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  • Saccharomyces cerevisiae  (57)
  • Arabidopsis  (50)
  • Springer  (107)
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  • 1
    ISSN: 1572-8773
    Schlagwort(e): EPR ; Saccharomyces cerevisiae ; uptake ; vanadate ; vanadyl
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie
    Notizen: Abstract Vanadium uptake by whole cells and isolated cell walls of the yeast Saccharomyces cerevisiae was studied. When orthovanadate was added to wild-type S. cerevisiae cells growing in rich medium, growth was inhibited as a function of the VO4 3- concentration and the growth was completely arrested at a concentration of 20 mM of VO4 3- in YEPD. Electron paramagnetic resonance (EPR) spectroscopy was used to obtain structural and dynamic information about the cell-associated paramagnetic vanadyl ion. The presence of EPR signals indicated that vanadate was reduced by whole cells to the vanadyl ion. On the contrary, no EPR signals were detected after interaction of vanadate with isolated cell walls. A ‘mobile’ and an ‘immobile’ species associated in cells with small chelates and with macromolecular sites, respectively, were identified. The value of rotational correlation time τ r indicated the relative motional freedom at the macromolecular site. A strongly ‘immobilized’ vanadyl species bound to polar sites mainly through coulombic attractions was detected after interaction of VO2+ ions with isolated cell walls.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 2
    Digitale Medien
    Digitale Medien
    Springer
    Cellular and molecular life sciences 52 (1996), S. 1130-1135 
    ISSN: 1420-9071
    Schlagwort(e): Saccharomyces cerevisiae ; mitochondria ; mRNA-specific translational activation ; synthetic genes ; gene regulation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Abstract Mitochondrial gene expression in yeast,Saccharomyces cerevisiae, depends on translational activation of individual mRNAs by distinct proteins encoded in the nucleus. These nuclearly coded mRNA-specific translational activators are bound to the inner membrane and function to mediate the interaction between mRNAs and mitochondrial ribosomes. This complex system, found to date only in organelles, appears to be an adaptation for targeting the synthesis of mitochondrially coded integral membrane proteins to the membrane. In addition, mRNA-specific translational activation is a rate-limiting step used to modulate expression of at least one mitochondrial gene in response to environmental conditions. Direct study of mitochondrial gene regulation and the targeting of mitochondrially coded proteins in vivo will now be possible using synthetic genes inserted into mtDNA that encode soluble reporter/passenger proteins.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 3
    Digitale Medien
    Digitale Medien
    Springer
    Cellular and molecular life sciences 52 (1996), S. 1033-1041 
    ISSN: 1420-9071
    Schlagwort(e): Ubiquitin ; yeast ; Saccharomyces cerevisiae ; Dictyostelium discoideum ; cytoskeleton ; mutants ; endocytosis ; actin ; myosin ; calmodulin
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Abstract Endocytosis is a general term that is used to describe the internalization of external and plasma membrane molecules into the cell interior. In fact, several different mechanisms exist for the internalization step of this process. In this review we emphasize the work on the actin-dependent pathways, in particular in the yeastSaccharomyces cerevisiae, because several components of the molecular machinery are identified. In this yeast, the analysis of endocytosis in various mutants reveals a requirement for actin, calmodulin, a type I myosin, as well as a number of other proteins that affect actin dynamics. Some of these proteins have homology to proteins in animal cells that are believed to be involved in endocytosis. In addition, the demonstration that ubiquitination of some cell surface molecules is required for their efficient internalization is described. We compare the actin, myosin and ubiquitin requirements for endocytosis with recent results found studying these processes usingDictyostelium discoideum and animal cells.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 4
    Digitale Medien
    Digitale Medien
    Springer
    Cellular and molecular life sciences 52 (1996), S. 1111-1116 
    ISSN: 1420-9071
    Schlagwort(e): Mitochondria ; mitochondrial inheritance ; cytoskeleton ; Saccharomyces cerevisiae ; Schizosaccharomyces pombe ; membrane proteins ; organelle movement ; mitochondrial morphology
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Abstract Mechanisms mediating the inheritance of mitochondria are poorly understood, but recent studies with the yeastsSaccharomyces cerevisiae andSchizosaccharomyces pombe have begun to identify components that facilitate this essential process. These components have been identified through the analysis of conditional yeast mutants that display aberrant mitochondrial distribution at restrictive conditions. The analysis of these mutants has uncovered several novel proteins that are localized either to cytoskeletal structures or to the mitochondria themselves. Many mitochondrial inheritance mutants also show altered mitochondrial morphology and defects in maintenance of the mitochondrial genome. Although some inheritance components and mechanisms appear to function specifically in certain types of cells, other conserved proteins are likely to mediate mitochondrial behavior in all eukaryotic cells.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 5
    Digitale Medien
    Digitale Medien
    Springer
    Cellular and molecular life sciences 52 (1996), S. 1148-1157 
    ISSN: 1420-9071
    Schlagwort(e): Saccharomyces cerevisiae ; mitochondrial ribosomes ; peptidyl transferase ; Varl ribosomal protein ; gene relocation ; posttranscriptional rRNA modification
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Abstract Mitochondria posses their own ribosomes responsible for the synthesis of a small number of proteins encoded by the mitochondrial genome. In yeast,Saccharomyces cerevisiae, the two ribosomal RNAs and a single ribosomal protein, Varl, are products of mitochondrial genes, and the remaining approximately 80 ribosomal proteins are encoded in the nucleus. The mitochondrial translation system is dispensable in yeast, providing an excellent experimental model for the molecular genetic analysis of the fundamental properties of ribosomes in general as well as adaptations required for the specialized role of ribosomes in mitochondria. Recent studies of the peptidyl transferase center, one of the most highly conserved functional centers of the ribosome, and the Varl protein, an unusual yet essential protein in the small ribosomal subunit, have provided new insight into conserved and divergent features of the mitochondrial ribosome.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 6
    ISSN: 1432-2048
    Schlagwort(e): Arabidopsis ; Gravitropism ; phyB-1 ; Phytochrome
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Hypocotyls of dark-grown seedlings of Ara bidosis thaliana exhibit a strong negative gravitropism, which is reduced by red and also by long-wavelength, far-red light treatments. Light treatments using phytochrome A (phyA)- and phytochrome B (phyB)-deficient mutants showed that this response is controlled by phyB in a red/far-red reversible way, and by phyA in a non-reversible, very-low-fluence response. Crosses of the previously analyzed phyB-1 allele (in the ecotype Landsberg erecta background) to the ecotype Nossen wild-type (WT) background resulted in a WT-like negative gravitropism in darkness, indicating that the previously described gravitropic randomization observed with phyB-1 in the dark is likely due to a second mutation independent of that in the PHYB gene.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 7
    ISSN: 1432-2048
    Schlagwort(e): Arabidopsis ; Chlorophyll biosynthesis ; Plastid ; Porphobilinogen deaminase (purification, subcellular location)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract A recombinant plasmid, pArab8, harbouring the cDNA encoding the mature form of the tetrapyrrole synthesis enzyme porphobilinogen deaminase (EC 4.3.1.8; also known as hydroxymethylbilane synthase) from Arabidopsis thaliana (L.) Heynh. has been constructed, and used to transform Escherichia coli. The porphobilinogen deaminase protein from Arabidopsis was overexpressed in this strain, and purified to homogeneity (3000-fold) with a yield of 20%. Antibodies were raised against the purified plant enzyme, and used in Western blot analysis, immunoprecipitation of enzyme activity and immuno-gold electron microscopy. The results indicate that the enzyme is confined to plastids in both leaves and roots. The implications of this finding for plant tetrapyrrole synthesis are discussed.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 8
    ISSN: 1432-2048
    Schlagwort(e): Arabidopsis ; Amino acid transport systems ; Gene mapping ; Proline (uptake kinetics)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The raz1 mutant of Arabidopsis thaliana (L.) Heynh. has been selected as resistant to the toxic proline analogue, azetidine-2-carboxylic acid (2AZ). Seedlings of the mutant tolerated fivefold higher concentrations of 2AZ (ED50 = 0.25 mM) than the wild-type seedlings (ED50 = 0.05 mM). The mutant gene was found to be semi-dominant and the corresponding RAZ1 locus was mapped on chromosome 5 at 69.6±1.8 cM. The resistance to 2AZ could be fully and exclusively accounted for by the lower uptake rate of the proline analogue in the mutant. The influx of L-proline in roots of wild-type seedlings could be dissected into two components: (i) a component with a high affinity and a low capacity for l-proline (K m≈20 gmM, V max≈60 nmol·(g FW)-1·h-1) and also a high affinity for L-2AZ (K i≈40 μM) and (ii) a low-affinity, high-capacity component (K m≈5 mM: V max = 1300 nmol·(g FW)-1·h-1). Clearly, the raz1 mutation affects the activity of a high-affinity transporter, because the high-affinity uptake of proline in the mutant was at least fivefold lower than in the wild-type, whereas the low-affinity uptake was unchanged.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 9
    ISSN: 1432-2048
    Schlagwort(e): Arabidopsis ; Cell axialization ; MONOPTEROS gene ; PIN-FORMED gene ; Polar auxin transport ; Vascular development
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract In the embryo of Arabidopsis thaliana (L.) Heynh., formation of the hypocotyl/root axis is initiated at the early-globular stage, recognizable as oriented expansion of formerly isodiametric cells. The process depends on the activity of the gene MONOPTEROS (MP); mp mutant embryos fail to produce hypocotyl and radicle. We have analyzed the morphology and anatomy of mp mutant plants throughout the Arabidopsis life cycle. Mutants form largely normal rosettes and root systems, but inflorescences either fail to form lateral flowers or these flowers are greatly reduced. Furthermore, the auxin transport capacity of inflorescence axes is impaired and the vascular strands in all analyzed organs are distorted. These features of the mutant phenotype suggest that the MP gene promotes cell axialization and cell file formation at multiple stages of plant development.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 10
    ISSN: 1432-0983
    Schlagwort(e): Key words Omnipotent suppression ; Microtubules ; Respiratory deficiency ; Saccharomyces cerevisiae
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  SUP35 and SUP45 genes determine the accuracy of translation at the stage of termination. We present indirect evidence indicating that these genes may also control some cellular process mediated by microtubules. A majority of sup35 and sup45 suppressor mutations confer supersensitivity to benomyl, the drug which de-polymerizes microtubules. In addition, data correlating phenotypic manifestations of sup45 suppressor mutations, involving sensitivity to benomyl, respiratory deficiency and a suppressor effect, are also presented.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 11
    ISSN: 1432-0983
    Schlagwort(e): Key words D-ribulose-5-phosphate 3-epimerase ; D-ribose-5-phosphate ketol-isomerase ; Pentose-phosphate pathway ; Saccharomyces cerevisiae
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract We have cloned and characterized the two remaining unknown genes of the non-oxidative part of the pentose-phosphate pathway of Saccharomyces cerevisiae encoding the enzymes D-ribulose-5-phosphate 3-epimerase (Rpe1p) and D-ribose-5-phosphate ketol-isomerase (Rki1p). Rpe1p has an unexpected high specific activity of 2148 mU × (mg protein)–1 in crude extracts. Deletion mutants of RPE1 show no enzyme activity and are unable to grow on D-xylulose. Unexpectedly, haploid rki1 deletion mutants are not viable. Functional expression of RKI1 was demonstrated following an increase of gene dosage in the haploid rki1 deletion mutant, which restored viability and specific D-ribose-5-phosphate ketol-isomerase activity. Both enzymes show high similarity to the deduced protein sequences of various open reading frames, expressed sequence tags or cDNAs from different organisms.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 12
    Digitale Medien
    Digitale Medien
    Springer
    Current genetics 30 (1996), S. 461-468 
    ISSN: 1432-0983
    Schlagwort(e): Keywords DNA repair ; Methylation damage ; Epistasis analysis ; Saccharomyces cerevisiae
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  The major genotoxicity of methyl methanesulfonate (MMS) is due to the production of a lethal 3-methyladenine (3MeA) lesion. An alkylation-specific base-excision repair pathway in yeast is initiated by a Mag1 3MeA DNA glycosylase that removes the damaged base, followed by an Apn1 apurinic/ apyrimidinic endonuclease that cleaves the DNA strand at the abasic site for subsequent repair. MMS is also regarded as a radiomimetic agent, since a number of DNA radiation-repair mutants are also sensitive to MMS. To understand how these radiation-repair genes are involved in DNA methylation repair, we performed an epistatic analysis by combining yeast mag1 and apn1 mutations with mutations involved in each of the RAD3, RAD6 and RAD52 groups. We found that cells carrying rad6, rad18, rad50 and rad52 single mutations are far more sensitive to killing by MMS than the mag1 mutant, that double mutants were much more sensitive than either of the corresponding single mutants, and that the effects of the double mutants were either additive or synergistic, suggesting that post-replication and recombination-repair pathways recognize either the same lesions as MAG1 and APN1, or else some differ- ent lesions produced by MMS treatment. Lesions handled by recombination and post replication repair are not simply 3MeA, since over-expression of the MAG1 gene does not offset the loss of these pathways. Based on the above analyses, we discuss possible mechanisms for the repair of methylation damage by various pathways.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 13
    ISSN: 1432-0983
    Schlagwort(e): Biocontrol ; Secretion ; Chitinase ; Expression cloning ; Saccharomyces cerevisiae ; Trichoderma harzianum
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract A novel endochitinase agar-plate assay has been developed and used to identify 11 full-length cDNAs encoding endochitinase I (ENC I) from aTrichoderma harzianum cDNA library by expression in yeast. The 1473-bpchil cDNA encodes a 424-residue precursor protein including both a signal sequence and a propeptide. The deduced ENC I amino-acid sequence is homologous to other fungal and bacterial chitinases, and the enzyme cross-reacts with a polyclonal antiserum raised against chitinase A1 fromBacillus circulans. TheT. harzianum endochitinase I was secreted into the culture medium by the yeastSaccharomyces cerevisiae in a functionally active form. The purified recombinant enzyme had a molecular mass of 44 kDa, an isoelectric point of 6.3, a pH optimum of 7.0 and a temperature optimum of 20 °C.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 14
    ISSN: 1432-0983
    Schlagwort(e): Heat-shock response ; Multidrug resistance ; AP-1 homolog ; Saccharomyces cerevisiae
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract We have examined whether the stress-induced transcriptional activation ofYDR1/PDR5/STS1 is mediated by yAP-1 and yAP-2. Of the stresses examined, heat shock-induced, rapid and transient PDR5 expression became very low in ayap1 yap2 double-gene disruptant, indicating that the yAP proteins mediate the response. Similar results were obtained withSNQ2, a close homologue ofPDR5. A set of 5′-truncation derivatives of thePDR5 gene identified the region from −484 to −434 as being sufficient for the response. A sequence similar to the yAP-1 recognition element recently identified in the stress-responsive yeast genes was found in this region and in the 5′-flanking sequences ofSNQ2.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 15
    ISSN: 1432-0983
    Schlagwort(e): Key words Biocontrol ; Secretion ; Chitinase ; Expression cloning ; Saccharomyces cerevisiae ; Trichoderma harzianum
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  A novel endochitinase agar-plate assay has been developed and used to identify 11 full-length cDNAs encoding endochitinase I (ENC I) from a Trichoderma harzianum cDNA library by expression in yeast. The 1473-bp chi1 cDNA encodes a 424-residue precursor protein including both a signal sequence and a propeptide. The deduced ENC I amino-acid sequence is homologous to other fungal and bacterial chitinases, and the enzyme cross-reacts with a polyclonal antiserum raised against chitinase A1 from Bacillus circulans. The T. harzianum endochitinase I was secreted into the culture medium by the yeast Saccharomyces cerevisiae in a functionally active form. The purified recombinant enzyme had a molecular mass of 44 kDa, an isoelectric point of 6.3, a pH optimum of 7.0 and a temperature optimum of 20 °C.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 16
    ISSN: 1432-0983
    Schlagwort(e): Key words Cytochrome b ; Mutants ; Mitochondria ; Saccharomyces cerevisiae
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The nucleotide changes present in a group of five cytochrome b mit– mutants were analyzed at the sequence level. Two single-base changes were found: one (M10-152) generated a nonsense codon in the first exon while the other (M8-181) created a missense substitution in the second exon. The other mutants all have multiple (three) substitutions that either resulted in a missense mutation in a coding region (M17-162) or else changed nucleotides in the last intron of the gene, so blocking its excision (M6-200 and M8-53). The synthesis of mitochondrial polypeptides and the steady state concentration of the complex-III subunits were examined. The Rieske protein and the core-4 and core-5 subunits were much reduced in all mutants. Consequently the overall stability of complex III is very sensitive even to amino-acid substitutions in the cytochrome b protein. Mutant M8-53 provides direct evidence for the proposed role of the P9.1 stem in the core structure of the group-I type last intron of this gene.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 17
    ISSN: 1432-0983
    Schlagwort(e): Key wordsβ-glucosidase ; Candida wickerhamii ; Saccharomyces cerevisiae
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The yeast Candida wickerhamii exports a cell-associated β-glucosidase that is active against cellobiose and all soluble cellodextrins. Because of its unique ability to tolerate end-product inhibition by glucose, the bglB gene that encodes this enzyme was previously cloned and sequenced in this laboratory. Using several different promoters and constructs, bglB was expressed in the hosts Escherichia coli, Pichia pastoris, and Saccharomyces cerevisiae. Expression was initially performed in E. coli using either the lacZ or tac promoter. This resulted in intracellular expression of the BglB protein with the protein being rapidly fragmented. Secretion and glycosylation of active β-glucosidase was achieved with several different S. cerevisiae constructs utilizing either the adh1 or the gal1 promoter on 2-µ replicating plasmids. When either the invertase (Suc2) or the BglB secretion signal was used, BglB protein remained associated with the cell wall and appeared to be hyperglycosylated. Expression in P. pastoris was also examined to determine if higher activity and expression could be achieved in a yeast host that usually does not hyperglycosylate. Using the alcohol oxidase promoter in conjunction with either the pho1 or the α-factor secretion signal, the recombinant enzyme was successfully secreted and glycosylated in P. pastoris. However, levels of protein expression from the chromosomally integrated vector were insufficient to detect activity.
    Materialart: Digitale Medien
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  • 18
    Digitale Medien
    Digitale Medien
    Springer
    Current genetics 29 (1996), S. 227-233 
    ISSN: 1432-0983
    Schlagwort(e): Trichoderma reesei ; β-Glucosidase ; Cellulase ; Saccharomyces cerevisiae
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract An intronless form of thebgl1 gene encoding an extracellularβ-glucosidase fromTrichoderma reesei was expressed in the yeast Saccharomyces cerevisiae under the control of the yeast GAL 1 promoter. Transformation of a yeast strain with this vector resulted in transformants that produce and secrete activeβ-glucosidase into the growth medium. Additionally, active recombinantβ-glucosidase protein was shown to be localized predominantly in the periplasmic space by using ap-nitrophenylβ-D-glycoside hydrolysis assay against fractionated yeast cells. The apparent size of the recombinant enzyme was 10–15 kDa larger than that of the native form. Treatment of the recombinantβ-glucosidase with endoglycosidase-H indicated the apparent increase in size was due to N-linked glycosylation.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 19
    Digitale Medien
    Digitale Medien
    Springer
    Current genetics 29 (1996), S. 227-233 
    ISSN: 1432-0983
    Schlagwort(e): Key words  Trichoderma reesei ; β-Glucosidase ; Cellulase ; Saccharomyces cerevisiae
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract   An intronless form of the bgl1 gene encoding an extracellular β-glucosidase from Trichoderma reesei was expressed in the yeast Saccharomyces cerevisiae under the control of the yeast GAL1 promoter. Transformation of a yeast strain with this vector resulted in transformants that produce and secrete active β-glucosidase into the growth medium. Additionally, active recombinant β-glucosidase protein was shown to be localized predominantly in the periplasmic space by using a p-nitrophenyl β-D-glycoside hydrolysis assay against fractionated yeast cells. The apparent size of the recombinant enzyme was 10–15 kDa larger than that of the native form. Treatment of the recombinant β-glucosidase with endoglycosidase-H indicated the apparent increase in size was due to N-linked glycosylation.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 20
    ISSN: 1432-0983
    Schlagwort(e): Key words Adenine biosynthesis ; ade8-18 ; ade2 mutations ; Red/white colony color assay ; Saccharomyces cerevisiae
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract In the yeast Saccharomyces cerevisiae the ade2, and/or the ade1, mutation in the adenine biosynthetic pathway leads to the accumulation of a cell-limited red pigment, while epistatic mutations in the same pathway, i.e. ade8, preclude this phenomenon, resulting in normal white colonies. The shift in color from red to white (or vice versa) with a combination of appropriate wild-type and mutant alleles of the adenine-pathway genes has been widely utilized as a non-selective phenotype to visualise and quantify the occurrence of various genetic events such as recombination, conversion and aneuploidy. It has provided an invaluable tool for the study of gene dosage and plasmid stability. In competition experiments between disrupted ade2, ade8-18 transformants carrying either a functional or non-functional episomal ADE8 gene, we verified that white ade8 ade2 cells show a remarkable selective advantage over red ade2 cells, with important implications on the use of this assay for the monitoring of genetic events. The accumulation of the red pigment in ade2 cells is likely to be the cause for impaired growth in these cells.
    Materialart: Digitale Medien
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  • 21
    ISSN: 1432-072X
    Schlagwort(e): Key words Plasma membrane H+-ATPase ; Saccharomyces cerevisiae ; Low pH ; PMA1 gene expression ; PMA2 gene expression
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Cells of Saccharomyces cerevisiae grown in media with an initial pH of 2.5–6.0, acidified with a strong acid (HCl), exhibited the highest plasma membrane H+-ATPase-specific activity at an initial pH of 6.0. At a lower pH (above pH 2.5) ATPase activity (62–83% of the maximum level) still allowed optimal growth. At pH 2.5, ATPase activity was about 30% of the maximum value and growth was impaired. Quantitative immunoassays showed that the content of ATPase protein in the plasma membrane was similar across the entire pH range tested, although slightly lower at pH 2.5. The decrease of plasma membrane ATPase activity in cells grown at low pH was partially accounted for by its in vitro stability, which decreased sharply at pH below 5.5, although the reduction of activity was far below the values expected from in vitro measurements. Yeast growth under acid stress changed the pattern of gene expression observed at optimal pH. The level of mRNA from the essential plasma-membrane-ATPase-encoding gene PMA1 was reduced by 50% in cells grown at pH 2.5 as compared with cells grown at the optimal pH 5.0, although the content of ATPase in the plasma membrane was only modestly reduced. As observed in response to other kinds of stress, the PMA2 promoter at the optimal pH was up to eightfold more efficient in cells grown at pH 2.5, although it remained several hundred times less efficient than that of the PMA1 gene.
    Materialart: Digitale Medien
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  • 22
    Digitale Medien
    Digitale Medien
    Springer
    Journal of industrial microbiology and biotechnology 16 (1996), S. 102-109 
    ISSN: 1476-5535
    Schlagwort(e): Millet ; Pennisetum typhoides ; liquefaction ; saccharification ; baker's yeast ; Saccharomyces cerevisiae ; fermentation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Werkstoffwissenschaften, Fertigungsverfahren, Fertigung
    Notizen: Abstract A fermentation medium based on millet (Pennisetum typhoides) flour hydrolysate and a four-phase feeding strategy for fed-batch production of baker's yeast,Saccharomyces cerevisiae, are presented. Millet flour was prepared by dry-milling and sieving of whole grain. A 25% (w/v) flour mash was liquefied with a thermostable 1,4-α-d-glucanohydrolase (EC 3.2.1.1) in the presence of 100 ppm Ca2+, at 80°C, pH 6.1–6.3, for 1 h. The liquefied mash was saccharified with 1,4-α-d-glucan glucohydrolase (EC 3.2.1.3) at 55°C, pH 5.5, for 2 h. An average of 75% of the flour was hydrolysed and about 82% of the hydrolysate was glucose. The feeding profile, which was based on a model with desired specific growth rate range of 0.18–0.23 h−1, biomass yield coefficient of 0.5 g g−1 and feed substrate concentration of 200 g L−1, was implemented manually using the millet flour hydrolysate in test experiments and glucose feed in control experiments. The fermentation off-gas was analyzed on-line by mass spectrometry for the calculation of carbon dioxide production rate, oxygen up-take rate and the respiratory quotient. Off-line determination of biomass, ethanol and glucose were done, respectively, by dry weight, gas chromatography and spectrophotometry. Cell mass concentrations of 49.9–51.9 g L−1 were achieved in all experiments within 27 h of which the last 15 h were in the fedbatch mode. The average biomass yields for the millet flour and glucose media were 0.48 and 0.49 g g−1, respectively. No significant differences were observed between the dough-leavening activities of the products of the test and the control media and a commercial preparation of instant active dry yeast. Millet flour hydrolysate was established to be a satisfactory low cost replacement for glucose in the production of baking quality yeast.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 23
    Digitale Medien
    Digitale Medien
    Springer
    Molecular and cellular biochemistry 158 (1996), S. 121-124 
    ISSN: 1573-4919
    Schlagwort(e): osmotic stress ; Saccharomyces cerevisiae ; glycerol ; K+/Na+ ions ; osmoregulation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Abstract The intracellular level of Na+ and K+ of S. cerevisiae strain AB1375 revealed that under KCl as well as sorbitol stress, the cationic level was comparable to the level under no stress conditions. On the other hand, there was a sharp drop in the intracellular K+ content and increase in the Na+ content on addition of NaCl to the medium. However, the total cationic level was close to that under control conditions. In addition to changes in the cationic level, an enhanced production and accumulation of glycerol were also observed under osmotic stress. A regulatory mechanism co-ordinating the intracellular concentration of glycerol as well as Na+, K+ content under osmotic stress conditions has been proposed.
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  • 24
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 30 (1996), S. 419-426 
    ISSN: 1573-5028
    Schlagwort(e): HEMA ; Glu-tRNA reductase ; Arabidopsis ; porphyrin ; chlorophyll
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract 5-Aminolevulinic acid (ALA) is the universal precursor of tetrapyrroles (e.g., chlorophylls and hemes). In the chloroplasts of plants and in several eubacterial species ALA is formed in a two-step process known as the C5 pathway. In the first step, glutamyl-tRNA reductase (GluTR), converts glutamate of glutamyl-tRNA to glutamate 1-semialdehyde (GSA) which is rearranged to ALA by glutamate 1-semialdehyde-2,1-aminomutase (GSA-AM) in the second step. Since ALA formation is a limiting step in chlorophyll biosynthesis, GluTR, which is encoded by the HEMA gene in Arabidopsis thaliana plays a vital role in that biosynthesis. Here we report the occurrence of a second functional HEMA gene (HEMA2) in A. thaliana. This gene was isolated by screening a genomic library with a probe from HEMA1. The nucleotide sequence of the cDNA and the corresponding genomic DNA indicates that the Arabidopsis HEMA2 gene contains two short introns (285 bp and 159 bp). The deduced amino acid sequence predicts a HEMA2 protein of 530 amino acids with 79% identity to the HEMA1-encoded GluTR. The 5′-flanking sequence of the HEMA2 gene includes several motifs (e.g., GT-1 boxes, GATA motifs) similar to light-responsive regulatory elements found in light-inducible genes. Unlike the HEMA1 transcript, which is present in all parts of the plant, HEMA2 is expressed in low levels in roots and flowers. The presence of a second functional HEMA gene in Arabidopsis raises the possibility that two C5 pathways exist in chloroplasts.
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  • 25
    ISSN: 1573-5028
    Schlagwort(e): tobamovirus ; Cruciferae ; CRMV ; Arabidopsis
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The complete nucleotide sequence of Chinese rape mosaic virus has been determined. The virus is a member of the tobamovirus genus of plant virus and is able to infect Arabidopsis thaliana (L.) Heynh systemically. The analysis of the sequence shows a gene array that seems to be characteristic of crucifer tobamoviruses and which is slightly different from the one most frequently found in tobamoviruses. Based on gene organization and on comparisons of sequence homologies between members of the tobamoviruses, a clustering of crucifer tobamoviruses is proposed that groups the presently known crucifer tobamovirus into two viruses with two strains each. A name change of Chinese rape mosaic virus to oilseed rape mosaic virus is proposed.
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  • 26
    ISSN: 1573-5028
    Schlagwort(e): Arabidopsis ; gene expression ; hypersensitive response ; plant-pathogen interactions ; cell suspensions ; sulfotransferase
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract A cDNA clone (RaRO47) encoding a sulfotransferase (ST) has been isolated from Arabidopsis cell suspensions. The deduced polypeptide of 302 amino acids is highly related to plant flavonol sulfotrans-ferases (FSTs), characterized for the first time in Flaveria, and also to STs from animal tissue. The expression of the Arabidopsis ST gene(s) corresponding to RaR047 was examined during different developmental stages. It was found that, at the level of steady-state mRNA, expression of gene(s) encoding this ST was rapidly induced in the aerial parts of young seedlings, and during growth of Arabidopsis cell cultures. No expression could be detected in roots. Treatment of Arabidopsis seedlings with hormonal or stress-related compounds, showed that RaR047 mRNA accumulation was more particularly induced in response to salicylic acid and methyl jasmonate. Furthermore, in the leaves of mature plants or in cell suspensions, accumulation of RaR047 mRNA was observed upon infection with bacterial pathogens. This expression was observed preferentially in response to avirulent pathogens causing an hyper-sensitive reaction, as compared to virulent pathogens, which lead to disease.
    Materialart: Digitale Medien
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  • 27
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 31 (1996), S. 1079-1082 
    ISSN: 1573-5028
    Schlagwort(e): Arabidopsis ; photomorphogenesis ; phytochrome B ; signal transduction ; transgenic plants
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract In an attempt to identify domains directly involved in the signal transduction of phytochrome B (phyB), we over-expressed the achromophoric C-terminal half of phyB under control of the CaMV-35S promoter in transgenicArabidopsis. In three independent transgenic lines, we detected accumulation of the introduced protein of predicted size at levels higher than that of the endogenous phyB by immunoblot analysis. Although these transgenic plants did not show any phenotype in the dark, enhancement of the phyB-dependent inhibition of hypocotyl elongation and reduction of the phytochrome A (phyA)-dependent inhibition were observed.
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  • 28
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 32 (1996), S. 549-552 
    ISSN: 1573-5028
    Schlagwort(e): Arabidopsis ; cysteine tRNA genes ; genome organization ; in vitro transcription ; Nicotiana
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract We have recently characterized Nicotiana cytoplasmic (cyt) tRNAGCA Cys as novel UGA suppressor tRNA. Here we have isolated its corresponding (NtC1) and a variant (NtC2) gene from a genomic library of Nicotiana rustica. Both tRNACys genes are efficiently transcribed in HeLa cell nuclear extract and yield mature cyt tRNAsCys. Sequence analysis of the upstream region of the RAD51 single-copy gene of the Arabidopsis thaliana genome revealed a cluster of three tRNACys genes which have the same polarity and comprise highly similar flanking sequences. Of the three Arabidopsis tRNACys genes only one (i.e. AtC2) appears to code for a functional gene which exhibits an almost identical nucleotide sequence to NtC1. These are the first sequenced nuclear tDNAsCys of plant origin.
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  • 29
    ISSN: 1573-5028
    Schlagwort(e): MAP kinase ; osmotic stress ; Pisum sativum ; Saccharomyces cerevisiae ; signal transduction
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Previous analysis of the MAP kinase homologue from Pisum sativum (PsMAPK) revealed a potential MAP kinase motif homologous to that found in eukaryotic cdc2 kinases. Sequence comparison showed a 47% identity on amino acid sequence basis to the Saccharomyces cerevisiae Hog 1p MAP kinase involved in the osmoregulatory pathway. Under conditions of salt-stress aberrant morphology of a hog1 deletion mutant was completely restored and growth was partially restored by expression of the PsMAPK. This shows that PsMAPK is functionally active as a MAP kinase in S. cerevisiae. Comparison of PsMAPK with other kinases involved in osmosensitivity, showed a high degree of homology and implicates a possible role for PsMAPK in a P. sativum osmosensing signal transduction pathway.
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  • 30
    ISSN: 1573-5028
    Schlagwort(e): Arabidopsis ; DNA repair ; tolerance ; DNA damage ; thiamine biosynthesis
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract AnArabidopsis thaliana cDNA was isolated by complementation of theEscherichia coli mutant strain BW535 (xth, nfo, nth), which is defective in DNA base excision repair pathways. This cDNA partially complements the methyl methane sulfonate (MMS) sensitive phenotype of BW535. It also partially corrects the UV-sensitive phenothpe ofE. coli AB1886 (uvrA) and restores its ability to reactivate UV-irradiated λ phage. It has an insert of ca. 1.3 kb with an open reading frame of 1047 bp (predicting a protein with a molecular mass of 36 kDa). This cDNA presents a high homology to a stress related gene from two species ofFusarium (sti35) and to genes whose products participate in the thiamine biosynthesis pathway,THI4, fromSaccharomyces cerevisiae andnmt2 fromSchizosaccharomyces pombe. TheArabidopsis predicted polypeptide has homology to several protein motifs: amino-terminal chloroplast transit peptide, dinucleotide binding site, DNA binding and bacterial DNA polymerases. The auxotrophy for thiamine in the yeastthi4::URA3 disruption strain is complemented by theArabidopsis gene. Thus, the cloned gene, namedthi1, is likely to function in the biosynthesis of thiamine in plants. The data presented in this work indicate thatthi1 may also be involved in DNA damage tolerance in plant cells.
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  • 31
    ISSN: 1573-5028
    Schlagwort(e): Arabidopsis ; calcium ; calmodulin ; gene family ; kinase ; phylogeny
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract A family of calcium-responsive protein kinases is abundant in plant cell extracts but has not been identified in animals and fungi. These enzymes have a unique structure consisting of a protein kinase catalytic domain fused to carboxy-terminal autoregulatory and calmodulin-like domains. In this report, we present the amino acid sequences for eight new Arabidopsis cDNA clones encoding isoforms of this enzyme. Three isoforms were expressed as fusion proteins in Escherichia coli and exhibited calcium-stimulated protein kinase activity. We propose CPK as the gene designation for this family of enzymes and describe a phylogenetic analysis for all known isoforms.
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  • 32
    ISSN: 1573-5028
    Schlagwort(e): Arabidopsis ; PP2A regulatory subunit ; serine/threonine phosphatase
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract We have indentified a novel gene (AtBβ) encoding a previously uncharacterized isoform of the B regulatory subunit of the type 2A serine/threonine protein phosphatase (PP2A) of Arabidopsis, and show that mRNA derived from the AtBβ gene accumulates in all Arabidopsis organs. In addition, we examined the expression of the three genes encoding the A regulatory subunit of Arabidopsis PP2A and show these genes are expressed in all organs as well. Taken together, our results suggest a myriad of PP2A subunit combinations, possibly with distinct substrate specificities, may occur within each Arabidopsis cell.
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  • 33
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 32 (1996), S. 275-302 
    ISSN: 1573-5028
    Schlagwort(e): Arabidopsis ; biotechnology ; ClpAP protease ; protein degradation ; 20S and 26S proteasome ; ubiquitin
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Proteolysis is essential for many aspects of plant physiology and development. It is responsible for cellular housekeeping and the stress response by removing abnormal/misfolded proteins, for supplying amino acids needed to make new proteins, for assisting in the maturation of zymogens and peptide hormones by limited cleavages, for controlling metabolism, homeosis, and development by reducing the abundance of key enzymes and regulatory proteins, and for the programmed cell death of specific plant organs or cells. It also has potential biotechnological ramification in attempts to improve crop plants by modifying protein levels. Accumulating evidence indicates that protein degradation in plants is a complex process involving a multitude of proteolytic pathways with each cellular compartment likely to have one or more. Many of these have homologous pathways in bacteria and animals. Examples include the chloroplast ClpAP protease, vacuolar cathepsins, the KEX2-like proteases of the secretory system, and the ubiquitin/26S proteasome system in the nucleus and cytoplasm. The ubiquitin-dependent pathway requires that proteins targeted for degradation become conjugated with chains of multiple ubiquitins; these chains then serve as recognition signals for selective degradation by the 26S proteasome, a 1.5 MDa multisubunit protease complex. The ubiquitin pathway is particularly important for developmental regulation by selectively removing various cell-cycle effectors, transcription factors, and cell receptors such as phytochrome A. From insights into this and other proteolytic pathways, the use of phosphorylation/dephosphorylation and/or the addition of amino acid tags to selectively mark proteins for degradation have become recurring themes.
    Materialart: Digitale Medien
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  • 34
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 30 (1996), S. 565-575 
    ISSN: 1573-5028
    Schlagwort(e): Arabidopsis ; cell cycle ; cell division ; cyclins ; polymerase chain reaction ; yeast complementation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Cyclins, a large family of proteins, are the regulatory subunits of cyclin-dependent protein kinases that are essential activators of cell cycle progression in eukaryotes. Here we report isolation of a new cyclin cDNA (cyc1bAt) from Arabidopsis cDNA libraries using polymerase chain reaction amplified cyclin-box sequences as probes. The deduced amino acid sequence of the isolated cDNA showed the highest sequence similarity with mitotic cyclins. However, the nucleotide and predicted amino acid sequence of cyc1bAt is different from five other mitotic-like cyclins that have recently been isolated from the same system, indicating that it is a new mitotic-like cyclin. These results, together with previous reports, suggest that there are at least six different mitotic-like cyclins in Arabidopsis. Expression of cyc1bAt in yeast G1 cyclin-minus mutant (DL1) rescued the cyclin-minus phenotype, demonstrating that plant mitotic-like cyclin can complement cyclin function in yeast. Analysis of expression of cyc1bAt in different tissues by reverse transcription-polymerase chain reaction using primers corresponding to a unique region of the cDNA showed that cyc1bAt is differentially expressed in different tissues with highest expression in flowers and no detectable expression in leaves.
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  • 35
    ISSN: 1573-5028
    Schlagwort(e): Arabidopsis ; metallothionein-like genes ; MT2 ; smt ; Synechococcus PCC 7942
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Zn2+ proteins pervade metabolism and are essential for gene expression. However, no proteins have been ascribed the central roles of Zn2+ donation to, or removal from, metalloproteins, or Zn2+ storage in vegetative plant tissue. In animals, such functions have been proposed for metallothioneins. Plants contain multiple metallothionein-like genes but their predicted products, which differ significantly from animal metallothioneins, remain to be isolated from vegetative tissue and their roles are uncertain. The type 2 metallothionein-like gene from Arabidopsis, MT2, was expressed under the control of Zn2+-responsive elements derived from the cyanobacterial metallothionein divergon, smt. Zn2+-dependent expression of MT2 transcripts in Synechococcus PCC 7942 was confirmed by northern analysis. The Arabidopsis MT2 gene partly complemented Zn2+ hypersensitivity in mutants of Synechococcus PCC 7942 which are functionally deficient in an endogenous Zn2+-metallothionein gene, smtA. MT2 was also expressed as a recombinant fusion protein in Escherichia coli, purified and shown to bind Zn2+ in vitro. The mean pH of half displacement of Zn2+ from MT2 was estimated to be 5.05. This suggests that MT2 has a greater affinity for Zn2+ than phytochelatins. The results presented here reveal that MT2 is capable of binding Zn2+ in vitro, conferring tolerance to elevated [Zn2+] in vivo within cyanobacteria and is likely to compete with other polypeptides for cellular Zn2+ in planta.
    Materialart: Digitale Medien
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  • 36
    ISSN: 1573-5028
    Schlagwort(e): Arabidopsis ; DNA-binding activity ; HSE (heat shock element) ; HSF (heat shock factor) ; tobacco (Nicotiana tabacum)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The DNA-binding activity of a tobacco heat shock factor (HSF) was induced by heat treatment (37–40 °C) of a cell-free extract that contained extra-nuclear fraction, but not in an extract of isolated nuclei. These observations suggest that an inactive form of HSF can directly recognize and transduce the heat shock signal and that such transduction requires components of the extranuclear fraction. Addition of ATP or of most other nucleoside triphosphates reduced the binding of the HSF to the heat shock element (HSE) in the same extract, and removal of ATP by dialysis from the extract restored the ability of the HSF to bind to DNA. The restored activity of the HSF could be eliminated again by a second addition of ATP. Our observations provide the first example of the involvement of ATP in the regulation of the reversible changes in HSF that control its ability to bind to HSEs in a cell-free extract.
    Materialart: Digitale Medien
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  • 37
    ISSN: 1573-5028
    Schlagwort(e): Arabidopsis ; glutathione ; heavy metals ; phytochelatin ; Schizosaccharomyces pombe
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Glutathione is essential for protecting plants from a range of environmental stresses, including heavy metals where it acts as a precursor for the synthesis of phytochelatins. A 1658 bp cDNA clone for glutathione synthetase (gsh2) was isolated fromArabidopsis thaliana plants that were actively synthesizing glutathione upon exposure to cadmium. The sequence of the clone revealed a protein with an estimated molecular mass of 53858 Da that was very similar to the protein from higher eukaryotes, was less similar to the gene from the fission yeast,Schizosaccharomyces pombe, and shared only a small region of similarity with theEscherichia coli protein. A 4.3 kbSstI fragment containing the genomic clone for glutathione synthetase was also isolated and sequenced. A comparison of the cDNA and genomic sequences revealed that the gene was composed of twelve exons. When theArabidopsis cDNA cloned in a special shuttle vector was expressed in aS. pombe mutant deficient in glutathione synthetase activity, the plant cDNA was able to complement the yeast mutation. Glutathione synthetase activity was measurable in wild-type yeast cells, below detectable levels in thegsh2 - mutant, and restored to substantial levels by the expression of theArabidopsis cDNA. TheS. pombe mutant expressing the plant cDNA had near wild type levels of total cellular thiols,109Cd2+ binding activity, and cadmium resistance. Since theArabidopsis cDNA was under control of a thiamine-repressible promoter, growth of the transformed yeast on thiamine-free medium increased expression of the cDNA resulting in increases in cadmium resistance.
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  • 38
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 32 (1996), S. 727-734 
    ISSN: 1573-5028
    Schlagwort(e): Arabidopsis ; essential amino acids ; lysine ; threonine ; transgenic plants
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract To study the regulation of lysine and threonine metabolism in plants, we have transformed Arabidopsis thaliana with chimeric genes encoding the two bacterial enzymes dihydrodipicolinate synthase (DHPS) and aspartate kinase (AK). These bacterial enzymes are much less sensitive to feedback inhibition by lysine and threonine than their plant counterparts. Transgenic plants expressing the bacterial DHPS overproduced lysine, but lysine levels were quite variable within and between transgenic genotypes and there was no direct correlation between the levels of free lysine and the activity of DHPS. The most lysine-overproducing plants also exhibited abnormal phenotypes. However, these phenotypes were detected only at early stages of plant growth, while at later stages, new buds emerged that looked completely normal and set seeds. Wild-type plants exhibited relatively high levels of free threonine, suggesting that in Arabidopsis AK regulation may be more relaxed than in other plants. This was also supported by the fact that expression of the bacterial AK did not cause any dramatic elevation in this amino acid. Yet, the relaxed regulation of threonine synthesis in Arabidopsis was not simply due to a reduced sensitivity of the endogenous AK to feedback inhibition by lysine and threonine because growth of wild-type plants, but not of transgenic plants expressing the bacterial AK, was arrested in media containing these two amino acids. The present results, combined with previous studies from our laboratory, suggest that the regulation of lysine and threonine metabolism is highly variable among plant species and is subject to complex biochemical, physiological and environmental controls. The suitability of these transgenic Arabidopsis plants for molecular and genetic dissection of lysine and threonine metabolism is also discussed.
    Materialart: Digitale Medien
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  • 39
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 32 (1996), S. 797-808 
    ISSN: 1573-5028
    Schlagwort(e): Arabidopsis ; Cucumis ; cytokinin ; noncoding RNA
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The CR20 gene was identified as a cytokinin-repressed gene in excised cotyledons of cucumber. We determined the sequences of some CR20 cDNAs with different structures and sequenced genomic clones for CR20. This gene consisted of three exons, and there were at least three types of transcript, which seemed to be generated by alternative splicing of the second intron. None of the CR20 transcripts included a long open reading frame (ORF). We isolated a cDNA of Arabidopsis thaliana with cucumber CR20 cDNA as a probe. This cDNA for a gene designated AtCR20-1 also lacked a long ORF. A region of 180 nucleotides was conserved in the CR20 RNA of cucumber and the AtCR20-1 RNA of Arabidopsis, although the homology was relatively low when the entire sequences were compared. Each conserved region consisted of seven elements, and seems to form stable secondary structure. These suggest that CR20 RNA may function as an RNA that is not translated into a protein.
    Materialart: Digitale Medien
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  • 40
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 32 (1996), S. 915-922 
    ISSN: 1573-5028
    Schlagwort(e): Arabidopsis ; DNA sequence polymorphism ; ecotypes ; genetic similarity ; sequence comparison
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract DNA polymorphisms among Arabidopsis thaliana ecotypes are widely used as genetic markers in map-based cloning strategies. New PCR-based molecular markers do not only facilitate molecular mapping, but can also be used to obtain reliable sequence information for cladistic analyses. We have used CAPS (cleaved amplified polymorphic sequences) markers and a direct sequencing strategy to estimate genetic similarity among eighteen Arabidopsis ecotypes. Sequences at four loci, two from the nuclear and two from a non-nuclaar genome, were analysed. For each ecotype more than 1000pb of sequence information was obtained, and genetic similarity was calculated from a total of 35 polymorphic sites using a character-based approach. Divergence ranged from zero up to 50 discordant characters among the 72 characters defined by the polymorphisms. Separate calculations based on the nuclear and the non-nuclear sequences were performed and revealed a number of common features, including the existence of small clusters of very closely related ecotypes separated from each other by extensive sequence divergence. Our results provide information useful especially to investigators setting up crosses for chromosome landing strategies.
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  • 41
    ISSN: 1573-5028
    Schlagwort(e): Arabidopsis ; differential screening ; light-regulated gene expression ; MYB-related proteins ; transcription factors
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Seven different MYB-related genes have been isolated from a genomic Arabidopsis library with probes based on MYB DNA-binding motifs. The predicted amino acid sequence of these genes showed high similarity in the MYB domain but outside this region virtually no similarities were found. The set of MYB-related genes was used to identify differentially expressed genes following the transfer of etiolated seedlings to light. This differential screen resulted in the selection of the ATM4 gene which is induced by light within one hour of exposure of etiolated or dark-adapted seedlings.
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  • 42
    ISSN: 1432-2048
    Schlagwort(e): Arabidopsis ; ω-3 fatty acid desaturase ; Leaf cell maturity ; Linolenic acid ; Lipid transfer ; Mutant
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Trienoic fatty acids, namely α-linolenic acid and hexadecatrienoic acid, present in leaf lipids are produced by ω-3 fatty acid desaturases located in the endoplasmic reticulum and plastid membranes. The changes in the level of trienoic fatty acids during leaf maturation were investigated in wild-type plants of Arabidopsis thaliana (L.) Heynh. and in the fad7 mutant deficient in the activity of a plastid ω-3 desaturase. The levels of trienoic fatty acids increased in 26 °C- and 15 °C-grown wild-type plants with maturation of leaves. The increase in trienoic fatty acids was mainly due to galactolipids enriched in plastid membranes. In addition, the relative levels of trienoic fatty acids in major glycerolipids, including phospholipids enriched in the endoplasmic reticulum membranes, also increased with leaf maturation. By contrast, when the fad7 mutant was grown at 26 °C, the relative levels of trienoic fatty acids in individual lipids decreased with leaf maturation. The decreases in the levels of trienoic fatty acids, however, were alleviated when the fad7 mutant was grown at 15 °C. These results suggest that the plastid ω-3 desaturase plays a major role in increasing the levels of trienoic fatty acids with leaf maturation.
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  • 43
    ISSN: 1432-2048
    Schlagwort(e): Arabidopsis ; Calmodulin ; Gravitropism
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Calmodulin, a primary plant calcium receptor, is known to be intimately involved with gravitropic sensing and transduction. Using the calmodulin-binding inhibitors trifluoperazine, W7 and calmidazolium, gravitropic curvature of Arabidopsis thaliana (L.) Heynh, ecotype Landsberg, roots was separable into two phases. Phase I was detected at very low concentrations (0.01 μM) of trifluoperazine and calmidazolium, did not involve growth changes, accounted for about half the total curvature of the root and may represent the specific contribution of the cap to gravity sensing. Phase II commenced around 1.0 μM and involved inhibition of both growth and curvature. The agr-3 mutant exhibited a reduced gravitropic response and was found to lack phase I curvature, suggesting that the mutation alters either use or expression of calmodulin. The sequences of wild-type and agr-3 calmodulin (CaM-1) cDNAs, which are root specific were completely determined and found to be identical. Upon gravitropic stimulation, wild-type Arabidopsis seedlings increased calmodulin mRNA levels by threefold in 0.5 h. On the other hand, gravitropic stimulation of agr-3 decreased calmodulin mRNA accumulation. The possible basis of the two phases of curvature is discussed and it is concluded that agr-3 has a lesion located in a general gravity transmission sequence, present in many root cells, which involves calmodulin mRNA accumulation.
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  • 44
    ISSN: 1432-2048
    Schlagwort(e): Arabidopsis ; Ascorbate peroxidase (cytosolic, plastidial) ; Multigene family ; Oxidative stress ; Pisum
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract A second cytosolic ascorbate peroxidase (cAPX; EC 1.11.1.11) gene from Arabidopsis thaliana has been characterised. This second gene (designated APX1b) maps to linkage group 3 and potentially encodes a cAPX as closely related to that from other dicotyledonous species as to the other member of this gene family (Kubo et al, 1993, FEBS Lett 315: 313–317; here designated APX1a), which maps to linkage group 1. In contrast, the lack of sequence similarity in non-coding regions of the genes implies that they are differentially regulated. Under non-stressed conditions only APX1a is expressed. APX1b was identified during low-stringency probing using a cDNA coding for pea cAPX which, in turn, was recovered from a cDNA library by immunoscreening with an antiserum raised against tea plastidial APX (pAPX). No pAPX cDNAs were recovered, despite the antiserum displaying specificity for pAPX in Western blots.
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  • 45
    Digitale Medien
    Digitale Medien
    Springer
    Planta 198 (1996), S. 87-94 
    ISSN: 1432-2048
    Schlagwort(e): Arabidopsis ; Gravitropism ; High-gradient magnetic field ; Linum ; Root growth
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract High-gradient magnetic fields (HGMFs) were used to induce intracellular magnetophoresis of amyloplasts. The HGMFs were generated by placing a small ferromagnetic wedge into a uniform magnetic field or at the gap edge between two permanent magnets. In the vicinity of the tip of the wedge the dynamic factor of the magnetic field, ▽(H2/2), was about 109 Oe2 · cm−1, which subjected the amyloplasts to a force comparable to that of gravity. When roots of 2-d-old seedlings of flax (Linum usitatissimum L.) were positioned vertically and exposed to an HGMF, curvature away from the wedge was transient and lasted approximately 1 h. Average curvature obtained after placing magnets, wedge and seedlings on a 1-rpm clinostat for 2 h was 33 ± 5 degrees. Roots of horizontally placed control seedlings without rotation curved about 47 ± 4 degrees. The time course of curvature and changes in growth rate were similar for gravicurvature and for root curvature induced by HGMFs. Microscopy showed displacement of amyloplasts in vitro and in vivo. Studies with Arabidopsis thaliana (L.) Heynh. showed that the wild type responded to HGMFs but the starchless mutant TC7 did not. The data indicate that a magnetic force can be used to study the gravisensing and response system of roots.
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  • 46
    ISSN: 1432-2048
    Schlagwort(e): Antioxidant enzymes ; Arabidopsis ; Chilling stress ; Glutathione ; Hydrogen peroxide ; Lipid peroxidation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Chilling ofArabidopsis thaliana (L.) Heynh. callus tissue to 4 °C led to conditions of oxidative stress, as indicated by increased levels of the products of peroxidative damage to cell membranes. Cellular H2O2 was also observed to increase initially upon chilling but by day 8 cellular levels had declined to below control levels. Although levels of catalase activity remained similar to those in unchilled tissue, activity of ascorbate peroxidase increased between days 4 and 8 of chilling to 4 °C. In callus held at 23 °C, levels of reduced glutathione remained static whereas they rose in callus held at 4 °C. Levels of oxidised glutathione were initially low but increased significantly by day 4 in the chilled callus. At 23 °C, however, levels of oxidised glutathione remained low. Between days 1 and 3 at 4 °C, levels of glutathione reductase activity increased but by day 8 glutathione reductase activity was similar to that in cells held at 23 °C. Exposure of callus to abscisic acid at 23 °C also led to increased activities of ascorbate peroxidase and glutathione reductase.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 47
    ISSN: 1432-2048
    Schlagwort(e): Arabidopsis ; Female gametophyte ; Pollen ; Reproduction ; Spaceflight
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Reproductive development in Arabidopsis thaliana (L.) Heynh. cv. Columbia plants was investigated under spaceflight conditions on shuttle mission STS-51. Plants launched just prior to initiation of the reproductive phase developed flowers and siliques during the 10-d flight. Approximately 500 flowers were produced in total by the 12 plants in both the ground control and spaceflight material, and there was no significant difference in the number of flowers in each size class. The flower buds and siliques of the spaceflight plants were not morphologically different from the ground controls. Pollen viability tests immediately post-flight using fluorescein diacetate indicated that about 35% of the pollen was viable in the spaceflight material. Light-microscopy observations on this material showed that the female gametophytes also had developed normally to maturity. However, siliques from the spaceflight plants contained empty, shrunken ovules, and no evidence of pollen transfer to stigmatic papillae was found by light microscopy immediately post-flight or by scanning electron microscopy on fixed material. Short stamen length and indehiscent anthers were observed in the spaceflight material, and a film-like substance inside the anther that connected to the tapetum appeared to restrict the release of pollen from the anthers. These observations indicate that given appropriate growing conditions, early reproductive development in A. thaliana can occur normally under spaceflight conditions. On STS-51, reproductive development aborted due to obstacles in pollination or fertilization.
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  • 48
    ISSN: 1432-2048
    Schlagwort(e): Arabidopsis ; High-chlorophyll-fluorescence mutants ; Photosynthesis
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Thirty-four recessive photosynthetic mutants of the high-chlorophyll-fluorescence (hcf) phenotype have been isolated by screening 7700 M2 progenies of ethyl methane sulfonate-treated seeds ofArabidopsis thaliana. Most of the mutants isolated were found to be seedlinglethal, but could be grown on sucrose-supplemented media. Chlorophyll (Chl) fluorescence induction, absorption changes in the reaction-centre chlorophyll of PS I (P700) at 830 nm and Chla/Chlb ratios were recorded in order to probe the photosynthetic functions and to define the mutational lesion. These studies were complemented by immunoblot and Northern analyses which finally led to the classification of the mutants into six different groups. Four classes of mutants were affected in PS I, PS II (two different classes) or the intersystem electron-transport chain, respectively. A fifth mutant class was of pleiotropic nature and the sixth class comprised a Chlb-deficient mutant. Several of the mutants showed severe deficiencies in the levels of subunits of PS I, PS II or the cytochromeb 6/f complex. Thus the mutational lesions could be located precisely. Only one mutant was defective in the transcript patterns of some plastid-encoded photosynthesis genes. Hence most of the mutants isolated appear to be affected in translational and post-translational regulatory processes of thylakoid membrane biogenesis or in structural genes encoding constituent subunits of the thylakoid protein complexes.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 49
    ISSN: 1432-2048
    Schlagwort(e): Arabidopsis ; Calmodulin cDNA (5′ untranslated region) ; Complementary DNA ; Open reading frame
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract A cDNA clone, pAUK1, with an open reading frame (ORF) coding for a hypothetical 164-amino-acid protein was isolated from an Arabidopsis thaliana (L.) Heynh cDNA library. The clone was attached, tail to tail, to the 3′ end of A. thaliana hexokinase cDNA. An almost identical sequence had been previously described as the 5′ untranslated region (5′ UTR) of A. thaliana calmodulin cDNA (ACaM-2). Sequence comparison with three additional A. thaliana truncated cDNA clones which appear in a database (GenBank) supports the conclusion that pAUKl is identical to the 5′ UTR of ACaM-2 and that the 5′ UTR of ACaM-2 is an independent cDNA artificially linked to A. thaliana calmodulin cDNA.
    Materialart: Digitale Medien
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  • 50
    ISSN: 1617-4623
    Schlagwort(e): Key words Saccharomyces douglasii ; Saccharomyces cerevisiae ; CBP2 ; Mitochondria ; Pre-mRNA processing
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  In Saccharomyces cerevisiae the only known role of the CBP2 gene is the excision of the fifth intron of the mitochondrial cyt b gene (bI5). We have cloned the CBP2 gene from Saccharomyces douglasii (a close relative of S. cerevisiae). A comparison of the S. douglasii and S. cerevisiae sequences shows that there are 14% nucleotide substitutions in the coding region, with transitions being three times more frequent than transversions. At the protein level sequence identity is 87%. We have demonstrated that the S. douglasii CBP2 gene is essential for respiratory growth in the presence of a wild-type S. douglasii mitochondrial genome, but not in the presence of an intronless S. cerevisiae mitochondrial genome. Also the S. douglasii and S. cerevisiae CBP2 genes are completely interchangeable, even though the intron bI5 is absent from the S. douglasii mitochondrial genome.
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  • 51
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 250 (1996), S. 267-276 
    ISSN: 1617-4623
    Schlagwort(e): Pre-mRNA splicing ; Saccharomyces cerevisiae ; Suppressors ; prp24-1
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The temperature-sensitiveprp24-1 mutation defines a gene product required for the first step in pre-mRNA splicing. PRP24 is probably a component of the U6 snRNP particle. We have applied genetic reversion analysis to identify proteins that interact with PRP24. Spontaneous revertants of the temperaturesensitive (ts)prp24-1 phenotype were analyzed for those that are due to extragenic suppression. We then extended our analysis to screen for suppressors that confer a distinct conditional phenotype. We have identified a temperature-sensitive extragenic suppressor, which was shown by genetic complementation analysis to be allelic toprp21-1. This suppressor,prp21-2, accumulates pre-mRNA at the non-permissive temperature, a phenotype similar to that ofprp21-1. prp21-2 completely suppresses the splicing defect and restores in vivo levels of the U6 snRNA in theprp24-1 strain. Genetic analysis of the suppressor showed thatprp21-2 is not a bypass suppressor ofprp24-1. The suppression ofprp24-1 byprp21-2 is gene specific and also allele specific with respect to both the loci. Genetic interactions with other components of the pre-spliceosome have also been studied. Our results indicate an interaction between PRP21, a component of the U2 snRNP, and PRP24, a component of the U6 snRNP. These results substantiate other data showing U2–U6 snRNA interactions.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 52
    ISSN: 1617-4623
    Schlagwort(e): Saccharomyces douglasii ; Saccharomyces cerevisiae ; CBP2 ; Mitochondria ; Pre-mRNA processing
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract InSaccharomyces cerevisiae the only known role of theCBP2 gene is the excision of the fifth intron of the mitochondrialcyt b gene (bI5). We have cloned theCBP2 gene fromSaccharomyces douglasii (a close relative ofS. cerevisiae). A comparison of theS. douglasii andS. cerevisiae sequences shows that there are 14% nucleotide substitutions in the coding region, with transitions being three times more frequent than transversions. At the protein level sequence identity is 87%. We have demonstrated that theS. douglasii CBP2 gene is essential for respiratory growth in the presence of a wild-typeS. douglasii mitochondrial genome, but not in the presence of an intronlessS. cerevisiae mitochondrial genome. Also theS. douglasii andS. cerevisiae CBP2 genes are completely interchangeable, even though the intron bI5 is absent from theS. douglasii mitochondrial genome.
    Materialart: Digitale Medien
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  • 53
    ISSN: 1617-4623
    Schlagwort(e): Saccharomyces cerevisiae ; Nucleotide excision repair ; RAD14 ; XPA homologue
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract In the accompanying paper we demonstrated that endonuclease III-sensitive sites in theMATα andHMLα loci ofSaccharomyces cerevisiae are repaired by the Nucleotide Excision Repair (NER) pathway. In the current report we investigated the repair of endonuclease III sites, 6-4 photoproducts and cyclobutane pyrimidine dimers (CPDs) in arad14-2 point mutant and in arad14 deletion mutant. TheRAD14 gene is the yeast homologue of the human gene that complements the defect in cells from xeroderma pigmentosum (XP) patients belonging to complementation group A. In the point mutant we observed normal repair of endonuclease III sites (i.e. as wild type), but no removal of CPDs at theMATα andHMLα loci. Similar experiments were undertaken using the recently createdrad14 deletion mutant. Here, neither endonuclease III sites nor CPDs were repaired inMAT a orHMR a. Thus the point mutant appears to produce a gene product that permits the repair of endonuclease III sites, but prevents the repair of CPDs. Previously it was found that, in the genome overall, repair of 6-4 photoproducts was less impaired than repair of CPDs in the point mutant. The deletion mutant repairs neither CPDs nor 6-4 photoproducts in the genome overall. This finding is consistent with the RAD14 protein being involved in lesion recognition in yeast. A logical interpretation is that therad14-2 point mutant produces a modified protein that enables the cell to repair endonuclease III sites and 6-4 photoproducts much more efficiently than CPDs. This modified protein may aid studies designed to elucidate the role of the RAD14 protein in lesion recognition.
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    Standort Signatur Erwartet Verfügbarkeit
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  • 54
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 250 (1996), S. 614-625 
    ISSN: 1617-4623
    Schlagwort(e): Key words RCC1 ; Saccharomyces cerevisiae ; Serine/threonine protein kinase
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  The yeast PRP20 protein is homologous to the RCC1 protein of higher eukaryotes and is required for mRNA export and maintenance of nuclear structure. RCC1/PRP20 act as guanine nucleotide exchange factors for the nuclear Ras-like Ran/GSP1 proteins. In a search for prp20-10 allele-specific high-copy-number suppressors, the KSP1 locus, encoding a serine/threonine protein kinase was isolated. Ksp1p is a nuclear protein that is not essential for vegetative growth of yeast. Inactivation of the kinase activity by a mutation affecting the catalytic center of the Ksp1p eliminated the suppressing activity. Based on the isolation of a protein kinase as a high-copy-number suppressor, the phosphorylation of Prp20p was examined. In vivo labeling experiments showed that Prp20p is a phosphoprotein; however, deletion of the KSP1 kinase did not affect Prp20p phosphorylation.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 55
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 251 (1996), S. 69-74 
    ISSN: 1617-4623
    Schlagwort(e): Key words Flowering ; Arabidopsis ; Photoperiod
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  The phenotype caused by mutations that affect the timing of flowering in Arabidopsis thaliana has been most extensively analyzed in the Landsberg erecta (Ler) genetic background. In Ler, the late-flowering phenotype of FRIGIDA and mutations in LUMINIDEPENDENS is suppressed by the Ler allele of FLC. In this study, the interactions of nine mutations conferring late flowering with the FLC allele of the Columbia ecotype (FLC-Col), which does not suppress late flowering, were examined. The effect on flowering time of combining six of the mutations with FLC-Col was additive; plants containing FLC-Col with fd, gi, fwa, fha, ft, and fe flowered slightly later than plants containing these mutations with the Ler allele of FLC. In contrast, a synergistic effect was observed between FLC-Col and three mutations; fca, fpa, and fve plants became extremely late flowering when combined with FLC-Col. Maximum delay in flowering for the majority of the mutant strains required FLC-Col in a homozygous state, although for fpa and fe a single copy of FLC-Col allowed maximum lateness. In addition, the fd and fe mutations became more dominant in the presence of FLC-Col.
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  • 56
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 253 (1996), S. 111-117 
    ISSN: 1617-4623
    Schlagwort(e): Key words MEL gene ; α-galactosidase ; Saccharomyces cerevisiae ; Saccharomyces paradoxus
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  In order to study the molecular evolution of the yeasts grouped in the Saccharomyces sensu stricto species complex by analysis of the MEL gene family, we have cloned and sequenced two new species-specific MEL genes from Saccharomyces yeasts: S. paradoxus (MELp) and a Japanese Saccharomyces sp. (MELj). The clones were identified by sequence homology to the S. cerevisiae MEL1 gene. Both clones revealed an ORF of 1413 bp coding for a protein of 471 amino acids. The deduced molecular weights of the α-galactosidase enzymes were 52 767 for MELp and 52 378 for MELj. The nucleotide sequences of the MELp (EMBL accession no. X95505) and the MELj (EMBL accession no. X95506) genes showed 74.7% identity. The degree of identity of MELp to the MEL1 gene was 76.8% and to the S. pastorianus MELx gene, 75.7%. The MELj coding sequence was 75.1% identical to the MEL1 gene and 80.7% to the MELx gene. The data suggest that MEL1, MELj, MELp, and MELx genes are species-specific MEL genes. The strains studied each have only one MEL locus. The MELp gene is located on the S. paradoxus equivalent of S. cerevisiae chromosome X; the MELj gene was on the chromosome that comigrates with the S. cerevisiae chromosome VII/XV doublet and hybridizes to the S. cerevisiae chromosome XV marker HIS3.
    Materialart: Digitale Medien
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  • 57
    ISSN: 1617-4623
    Schlagwort(e): Key words DEAD-box protein ; DED1 ; RCC1 ; Saccharomyces cerevisiae ; SRM1
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  The Saccharomyces cerevisiae temperature-sensitive mutants srm1-1, mtr1-2 and prp20-1 carry alleles of a gene encoding a homolog of mammalian RCC1. In order to identify a protein interacting with RCC1, a series of suppressors of the srm1-1 mutation were isolated as cold-sensitive mutants and one of the mutants, designated ded1-21, was found to be defective in the DED1 gene. The double mutant, srm1-1 ded1-21, could grow at 35° C, but not at 37° C. A revertant of srm1-1 ded1-21 that became able to grow at 37° C acquired another mutation in the SRM1 gene, indicating the tight relationship between SRM1 and DED1. In all the rcc1 - strains examined, the amount of mutated SRM1 proteins was reduced or not detectable at the nonpermissive temperature. While mutated SRM1 protein was stabilized in all of the rcc1 - strains by the ded1-21 mutation, the ded1-21 mutation suppressed both srm1-1 and mtr1-2, but not the prp20-1 mutation, contrary to the previous finding that overproduction of the S. cerevisiae Ran homolog GSP1 suppresses prp20-1, but not srm1-1 or mtr1-2.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 58
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 251 (1996), S. 682-691 
    ISSN: 1617-4623
    Schlagwort(e): Saccharomyces cerevisiae ; Two-hybrid system ; Protein-protein interactions ; PKC1 pathway ; MAP kinase cascade
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The two-hybrid system for the identification of protein-protein interactions was used to screen for proteins that interact in vivo with theSaccharomyces cerevisiae Pkc1 protein, a homolog of mammalian protein kinase C. Four positive clones were isolated that encoded portions of the protein kinase Mkk1, which acts downstream of Pkc1p in thePKC1-mediated signalling pathway. Subsequently, Pkc1p and the otherPKC1 pathway components encoding members of a MAP kinase cascade, Bck1p (a MEKK), Mkk1p, Mkk2p (two functionally homologous MEKs), and Mpk1p (a MAP kinase), were tested pairwise for interaction in the two-hybrid assay. Pkc1p interacted specifically with small N-terminal deletions of Mkk1p, and no interaction between Pkc1p and any of the other known pathway components could be detected. Interaction between Pkc1p and Mkk1p, however, was found to be independent of Mkk1p kinase activity. Bck1p was also found to interact with Mkk1p and Mkk2p, and the interaction required only the predicted C-terminal catalytic domain of Mkk1p. Furthermore, we detected protein-protein interactions between two Bck1p molecules via their N-terminal regions. Finally, Mkk2p and Mpk1p also interacted in the two-hybrid assay. These results suggest that the members of thePKC1-mediated MAP kinase cascade form a complex in vivo and that Pkc1p is capable of directly interacting with at least one component of this pathway.
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    Standort Signatur Erwartet Verfügbarkeit
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  • 59
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 251 (1996), S. 707-715 
    ISSN: 1617-4623
    Schlagwort(e): Saccharomyces cerevisiae ; Flocculation ; Transcriptional regulation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract A flocculation gene was cloned from aSaccharomyces cerevisiae ATCC60715 genomic library, known to contain theFLO8 gene, on the basis of its ability to confer a flocculation phenotype on a non-flocculent strain. From a total of 11 130 clones, four clones sharing the several restriction fragments were isolated, suggesting that these were derived from the same locus. The results of integration mapping and disruption of the cloned gene indicated that this gene was theFLO8 gene. After disruption of theFLO8 gene, the strain lost its ability to flocculate. The DNA sequence of theFLO8 gene was determined. This gene includes a 2187-bp open reading frame that encodes a 729-amino acid protein. Computer analysis indicated that theFLO8 gene has a significant degree of homology with aS. cerevisiae chromosome V DNA sequence, but no homology with theFLO1 gene. The hydrophobicity profile of the putativeFLO8 gene product did not indicate the presence of any significantly hydrophobic regions. Southern analysis of theFLO8 gene present in various yeast strains indicated that theFLO8 gene is highly conserved in yeast strains having a variety of flocculation phenotypes and genotypes. Northern analysis revealed that the level ofFLO1 gene transcription is dependent on the rate of transcription of theFLO8 gene. These results suggest that theFLO8 gene mediates flocculation via transcriptional activation of theFLO1 gene.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 60
    ISSN: 1617-4623
    Schlagwort(e): Key words 18S rRNA transcription ; Plant mitochondrial promoters ; Arabidopsis ; Pea ; Potato
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  Transcription initiation sites for the mitochondrial 18S rRNA genes in the dicot plants Arabidopsis thaliana, potato and pea were identified by a combination of in vitro capping, primer extension and S-1 analyses. These promoters contain a nonanucleotide motif and an AT-rich sequence similar to many mRNA and tRNA promoters in dicot mitochondria. In Arabidopsis and potato, active promoters are located within 120 nucleotides upstream of the 18S rRNA genes, as in Oenothera. The nucleotide sequence in the corresponding region in pea mitochondria is well conserved, but is not used as promoter in this plant. Instead a novel promoter sequence is used that lies several hundred nucleotides upstream. These results show that rRNAs can be transcribed from the same promoter types as mRNAs and tRNAs in plant mitochondria. However, the sequence features presently attributed to plant mitochondrial promoters – the conserved nonanucleotide and the upstream AT-rich box – do not allow to deduce the presence of an active promoter from genomic sequence data alone.
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    Standort Signatur Erwartet Verfügbarkeit
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  • 61
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 252 (1996), S. 510-517 
    ISSN: 1617-4623
    Schlagwort(e): FK506 binding protein (FKBP) ; Immunophilins ; Tetratricopeptide repeat (TPR) ; Plant stress ; Arabidopsis
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract We have isolated clones of an Arabidopsis gene (ROF1, forrotamaseFKBP) encoding a high molecular weight member of the FK506 binding protein (FKBP) family. The deduced amino acid sequence of ROF1 predicts a 551-amino acid, 62 kDa polypeptide which is 44% identical to human FKBP59 — a 59 kDa FKBP which binds to the 90 kDa heat shock protein and is associated with inactive steroid hormone receptors. ROF1 contains three FKBP12-like domains in the N-terminal portion of the protein (in contrast to two domains in mammalian FKBP59), an internal repeat structure associated with protein-protein interactions (tetratricopeptide repeats), and a putative calmodulin binding domain near the C-terminal region of the protein. No sequences associated with protein translocation out of the cytosol were found in ROF1.ROF1 mRNA was found at equivalent low levels in light-grown roots, stems, and flowers and at slightly higher levels in leaves. The abundance ofROF1 mRNA increased several-fold under stress conditions such as wounding or exposure to elevated NaCl levels.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 62
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 250 (1996), S. 395-404 
    ISSN: 1617-4623
    Schlagwort(e): Saccharomyces cerevisiae ; DNase ; Eukaryotes ; Genetic recombination
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Site-specific endonucleases have been found in various eukaryotic organelles such as mitochondria, chloroplasts and nuclei. These endonucleases initiate site-specific or homologous gene conversion in mitochondrial and nuclear DNA. Here, we report a new site-specific endonuclease activity, Endo.SK1, identified in mitochondria of strain SK1, a homothallic diploid strain ofSaccharomyces cerevisiae. Nucleotide sequences around the Endo.SK1-cleavage sites are different from those of known yeast site-specific endonucleases. The Endo.SK1 activity is, at least partly, specified by a gene in the SK1-derived mitochondria. A novel feature of the Endo.SK1 activity is its inducibility: the endonuclease activity was induced by ca. 40-fold by transfer of cells from a glucose medium into an acetate medium, and was then repressed. This transient induction was independent of the ploidy level of the cells, and coincided with induction of fumarase, a mitochondrial enzyme involved in the TCA cycle. Co-induction and co-repression of the mitochondrial site-specific endonuclease activity and a respiration-related enzyme indicate that the endonuclease activity is regulated in response to physiological conditions, and suggest a possible role for the endonuclease in mitochondrial DNA metabolism.
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    Standort Signatur Erwartet Verfügbarkeit
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  • 63
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 250 (1996), S. 681-691 
    ISSN: 1617-4623
    Schlagwort(e): Arabidopsis ; GNOM gene ; Intragenic complementation ; Conserved regions ; YeastYEC2 gene
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract TheGNOM gene is required for pattern formation along the main body axis of the embryo in the flowering plantArabidopsis thaliana. Mutations in theGNOM gene alter the asymmetric division of the zygote and interfere with the formation of distinct apical-basal regions in the developing embryo. We have isolated theGNOM gene by positional cloning, characterised its structure and determined the molecular lesions in mutant alleles. Although the predicted 163 kDa GNOM protein has a conserved domain in common with the yeast secretory protein Sec7p, it is most closely related in size and overall similarity to the product of the yeastYEC2 gene, which is not essential for cell viability. Four fully complementinggnom alleles carry missense mutations in conserved regions, seven partially complementing alleles have premature stop codon mutations and two non-complementing alleles have splice-site lesions. Our results suggest that the GNOM protein acts as a complex of identical subunits and that partial complementation may involve low levels of full-length protein generated by inefficient translational read-through.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 64
    ISSN: 1617-4623
    Schlagwort(e): Saccharomyces cerevisiae ; Calcineurin ; MAP kinase cascade ; Pheromone-induced growth arrest ; Synthetic effect
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Saccharomyces cerevisiae mutants which exhibit phenotypes (calcium resistance and vanadate sensitivity) similar to those of calcineurin-deficient mutants were isolated. The mutants were classified into four complementation groups (crv1,2,3 and4).crv1 was allelic tocnb1, a mutation in the regulatory subunit of calcineurin. The nucleotide sequences ofCRV2 andCRV3 genes which complemented thecrv2 andcrv3 mutations, respectively, are identical to those ofBCK1/SLK1/SKC1/SSP31 andMPK1/SLT2, respectively, which are both involved in the MAP kinase cascade. A calcineurin-deletion mutation (Δcnb1), which by itself has no detectable effect on growth and morphology, enhanced some phenotypes (slow growth and morphological abnormality) ofcrv2 andcrv3 mutants. These phenotypes ofcrv2 andcrv3 mutants were partially suppressed by Ca2+ or by overproduction of the calcineurin subunits (Cmp2 and Cnb1). Like the calcineurin-deficient mutant,crv2 andcrv3 mutants were defective in recovery from α-factor-induced growth arrest. The defect in recovery of the Δcnb1 mutant was suppressed by overexpression ofMPK1. These results indicated that the calcineurin-mediated and the Mpk1- (Bck1-) mediated signaling pathways act in parallel to regulate functionally redundant cellular events important for growth.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 65
    Digitale Medien
    Digitale Medien
    Springer
    Protoplasma 194 (1996), S. 81-90 
    ISSN: 1615-6102
    Schlagwort(e): Arabidopsis ; Pollen ; Vegetative cytoplasm ; Ultrastructure
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary Ultrastructural changes of pollen cytoplasm during generative cell formation and pollen maturation inArabidopsis thaliana were studied. The pollen cytoplasm develops a complicated ultra-structure and changes dramatically during these stages. Lipid droplets increase after generative cell formation and their organization and distribution change with the developmental stage. Starch grains in amyloplasts increase in number and size during generative and sperm cell formation and decrease at pollen maturity. The shape and membrane system of mitochondria change only slightly. Dictyo-somes become very prominent, and numerous associated vesicles are observed during and after sperm cell formation. Endoplasmic reticulum appears extensively as stacks during sperm cell formation. Free and polyribosomes are abundant in the cytoplasm at all developmental stages although they appear denser at certain stages and in some areas. In mature pollen, all organelles are randomly distributed throughout the vegetative cytoplasm and numerous small particles appear. Organization and distribution of storage substances and appearance of these small particles during generative and sperm cell formation and pollen maturation are discussed.
    Materialart: Digitale Medien
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  • 66
    ISSN: 1615-6102
    Schlagwort(e): Arabidopsis ; Handedness ; Helical array ; Maize ; Microtubule organization
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary The orientation of cortical microtubules in plant cells has been extensively studied, in part because of their influence on the expansion of most plant cell types. Cortical microtubules are often arranged in helical arrays, which are well known to occur with a specific pitch as a function of development or experimental treatment; however, it is not known if the handedness of helical arrays can also be specified. We have studied the handedness of helical arrays by using Vibratome sectioning of maize primary roots and confocal microscopy of Arabidopsis primary roots. In cortical cells of maize roots, the helical array was found to have the same handedness at a given position, not only for the cells of a single root, but also for the cells of more than one hundred roots examined. Quantification of angular distribution of apparent individual microtubules showed that defined regions of the root were composed of cells with highly uniform microtubule orientation. In the region between transverse and longitudinal microtubules (5–10.5 mm from the tip), the array formed a right-handed helix, and basal of cells with longitudinal microtubules (11.5–15 mm from the tip), the array formed a left-handed helix. Similarly, in epidermal cells of Arabidopsis roots right-handed helical arrays were found in the region between transverse and longitudinal microtubules. These results suggest that, in addition to the orientation of microtubules, the handedness of helical microtubule arrays is under cellular control.
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    Standort Signatur Erwartet Verfügbarkeit
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  • 67
    Digitale Medien
    Digitale Medien
    Springer
    Protoplasma 192 (1996), S. 178-188 
    ISSN: 1615-6102
    Schlagwort(e): Arabidopsis ; Root apical meristem development ; Dermatogen/calyptrogen histogen ; Spiral pattern ; Root cap ; Protoderm
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary We investigated the development of the root cap and protoderm inArabidopsis thaliana root tips.A. Thaliana roots have closed apical organization with the peripheral root cap, columella root cap and protoderm developing from the dermatogen/calyptrogen histogen. The columella root cap arises from columella initials. The initials for the peripheral root cap and protoderm are arranged in a collar and the initiation event for these cells occurs in a sequential pattern that is coordinated with the columella initials. The resulting root cap appears as a series of interconnected spiraling cones. The protoderm, in three-dimensions, is a cylinder composed of cell files made up of packets of cells. The number of cell files within the protoderm cylinder increases as the root ages from one to two weeks. The coordinated division sequence of the dermatogen/calyptrogen and the increase in the number of protoderm cell files are both features of post-embryonic development within the primary root meristem.
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    Standort Signatur Erwartet Verfügbarkeit
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  • 68
    Digitale Medien
    Digitale Medien
    Springer
    Protoplasma 195 (1996), S. 12-17 
    ISSN: 1615-6102
    Schlagwort(e): Protein histidine kinases ; Two-component systems ; Sensory photoreceptors ; Site-directed mutagenesis ; Arabidopsis ; Transgenic plants
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary The biochemical mechanism of phytochrome action is unknown. We have examined the proposal, based on sequence similarities to the sensor histidine kinase components of bacterial two-component signaling systems, that the phytochromes may be functional homologs of these kinases. Four amino acids, three highly conserved between the phytochrome and bacterial kinase molecules and the other, the histidine residue putatively the target of autophosphorylation, were changed singly in the oat phytochrome A sequence by in vitro site-directed mutagenesis, and the resultant mutant photo-receptor molecules were assayed for activity by overexpression in transgenic Arabidopsis. Three of the four mutant molecules retained activity equivalent to that of the unmutagenized parent sequence, whereas the fourth mutant could not be evaluated because of low expression. The data show that the former three mutagenized residues are not essential for phytochrome A function in transgenic Arabidopsis, but, because of the negative nature of the results, the possibility cannot be precluded that the photoreceptor functions as a protein kinase independent of these residues.
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  • 69
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 251 (1996), S. 69-74 
    ISSN: 1617-4623
    Schlagwort(e): Flowering ; Arabidopsis ; Photoperiod
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The phenotype caused by mutations that affect the timing of flowering inArabidopsis thaliana has been most extensively analyzed in the Landsbergerecta (Ler) genetic background. In Ler, the late-flowering phenotype ofFRIGIDA and mutations inLUMINIDEPENDENS is suppressed by the Ler allele ofFLC. In this study, the interactions of nine mutations conferring late flowering with theFLC allele of the Columbia ecotype (FLC-Col), which does not suppress late flowering, were examined. The effect on flowering time of combining six of the mutations withFLC-Col was additive; plants containingFLC-Col withfd, gi, fwa, fha, ft, andfe flowered slightly later than plants containing these mutations with theLer allele ofFLC. In contrast, a synergistic effect was observed betweenFLC-Col and three mutations;fca, fpa, andfve plants became extremely late flowering when combined withFLC-Col. Maximum delay in flowering for the majority of the mutant strains requiredFLC-Col in a homozygous state, although forfpa andfe a single copy ofFLC-Col allowed maximum lateness. In addition, thefd andfe mutations became more dominant in the presence ofFLC-Col.
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  • 70
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 251 (1996), S. 38-43 
    ISSN: 1617-4623
    Schlagwort(e): Saccharomyces cerevisiae ; Dynein ; Gene disruption ; cin8
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Cytoplasmic dynein is a multisubunit, microtubule-dependent motor enzyme that has been proposed to function in a variety of intracellular movements. As part of an effort to understand the evolution and the biological roles of cytoplasmic dynein, we have identified the first non-metazoan dynein light chain 1, SLC1, in the yeastSaccharomyces cerevisiae. The amino acid sequence of the SLC1 protein is similar to those of the human,Drosophila andCaenorhabditis cytoplasmic dynein light chains 1. TheSLC1 gene lies adjacent to theYAP2 (CAD1) transcription unit. TheSLC1 coding sequence is split by two introns and its mRNA is detectable throughout the cell cycle. Tetrad analysis of heterozygotes harboring aTRP insertion in theSLC1 coding region indicate thatSLC1 function is not essential for cell viability. Furthermore, we demonstrate that double mutants, defective forSLC1 and the kinesin-relatedCIN8 genes are non-lethal. The redundancy ofSLC1 function in yeast contrasts with the cell death caused by loss-of-function mutations in the dynein light chain 1 gene inDrosophila melanogaster.
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  • 71
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 251 (1996), S. 153-160 
    ISSN: 1617-4623
    Schlagwort(e): Key words hUBC9 ; Saccharomyces cerevisiae ; Ubc9p ; Yeast centromere proteins
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  Using a two-hybrid system, we cloned a human cDNA encoding a ubiquitin-conjugating enzyme (UBC), hUBC9, which interacts specifically with all three subunits of the Saccharomyces cerevisiae centromere DNA-binding core complex, CBF3. The hUBC9 protein shows highest homology to a new member of the UBC family: 54% identity to S. cerevisiae Ubc9p and 64% identity to Schizosaccharomyces pombe (Sp) hus5. Overexpression of hUBC9 partially suppresses a S. cerevisiae ubc9 temperature-sensitive mutation, indicating that the UBC9 gene family is also functionally conserved. Like hUBC9, Sphus5 also interacts specifically with all three subunits of the CBF3 complex. However, S. cerevisiae Ubc9p interacts only with the Cbf3p subunit (64 kDa) of the CBF3 complex, indicating the specificity of the interaction between S. cerevisiae Ubc9 and Cbf3p proteins. The function of Ubc9p in the G2/M phase of S. cerevisiae could be related to regulation of centromere proteins in chromosome segregation in mitosis. Therefore, the ubiquitination process and centromere function may be linked to chromosome segregation. We also provide further in vivo evidence that Mck1p, a protein kinase, is specifically associated with the centromere proteins Cbf2p and Cbf5p, which were previously shown to interact in vitro.
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  • 72
    ISSN: 1617-4623
    Schlagwort(e): Saccharomyces cerevisiae ; Cell cycle ; Dox-A2 ; NIN1 ; P91A ; SUN2
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract NIN1 is an essential gene for growth of the yeastSaccharomyces cerevisiae and was recently found to encode a component of the regulatory subunit of the 26S proteasome. Thenin1-1 mutant is temperature sensitive and its main defect is in G1/S progression and G2/M progression at non-permissive temperatures. One of the two multicopy suppressors ofnin1-1, SUN2 (SUppressor of Nin1-1), was found to encode a protein of 523 amino acids whose sequence is similar to those ofDrosophila melanogaster diphenol oxidase A2 and the mouse mast-cell Tum− transplantation antigen, P91A. The C-terminal half of Sun2p was found to be functional as Sun2p at 25° C, 30° C, and 34° C but not at 37° C. The open reading frame (ORF) of theDrosophila diphenol oxidase A2 gene (Dox-A2) was obtained from a lambda phage cDNA library using the polymerase chain reaction technique. TheDox-A2 ORF driven by theTDH3 promoter complemented the phenotype of a strain deleted forsun2. ThisDox-A2-dependent strain was temperature sensitive and accumulated dumb-bell-shaped cells, with an undivided nucleus at the isthmus, after temperature upshift. This morphology is similar to that ofnin1-1 cells kept at a restrictive temperature. These results suggest thatSUN2 is a functional counterpart ofDox-A2 and that these genes play a pivotal role in the cell cycle in each organism.
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  • 73
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 251 (1996), S. 176-185 
    ISSN: 1617-4623
    Schlagwort(e): Saccharomyces cerevisiae ; Cell cycle ; LTE1 ; CDC15 ; CDC14
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract LTE1 encodes a homolog of GDP-GTP exchange factors for the Ras superfamily and is required at low temperatures for cell cycle progression at the stage of the termination of M phase inSaccharomyces cerevisiae. We isolated extragenic suppressors which suppress the cold sensitivity oflte1 cells and confer a temperature-sensitive phenotype on cells. Cells mutant for the suppressor alone were arrested at telophase at non-permissive temperatures and the terminal phenotype was almost identical to that oflte1 cells at non-permissive temperatures. Genetic analysis revealed that the suppressor is allelic toCDC15, which encodes a protein kinase. Thecdc15 mutations thus isolated were recessive with regard to the temperature-sensitive phenotype and were dominant with respect to suppression oflte1. We isolatedCDC14 as a low-copy-number suppressor ofcdc15-rlt1.CDC14 encodes a phosphotyrosine phosphatase (PTPase) and is essential for termination of M phase. An extra copy ofCDC14 suppressed the temperature sensitivity ofcdc15-rlt1 cells, but not that ofcdc15-1 cells. In addition, some residues that are essential for the Cdc14 PTPase activity were found to be non-essential for the suppression. These results strongly indicate that Cdc14 possesses dual functions; PTPase activity is needed for one function but not for the other. We postulate that the cooperative action of Cdc14 and Cdc15 plays an essential role in the termination of M phase.
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  • 74
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 251 (1996), S. 261-266 
    ISSN: 1617-4623
    Schlagwort(e): Biotin ; Arabidopsis ; Complementation ; bioA
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract TheArabidopsis thaliana biotin auxotrophbio1 was rendered prototrophic by transformation with a chimeric transgene containing theEscherichia coli bioA gene driven by a constitutive promoter. ThebioA gene encodes the biotin biosynthetic enzyme 7,8-diaminopelargonic acid aminotransferase. Unlike the untransformed control plants, transgenic plants expressing the bacterial transgene synthesized biotin and grew to maturity without biotin-deficiency symptoms. These findings demonstrate thatbio1/bio1 mutant plants are defective in the gene encoding 7,8-diaminopelargonic acid aminotransferase.
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  • 75
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 251 (1996), S. 556-564 
    ISSN: 1617-4623
    Schlagwort(e): Key words cAMP ; Ca2+ ; Golgi apparatus ; Bud growth ; Saccharomyces cerevisiae
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  Biochemical and physiological studies have implicated cAMP and cAMP-dependent protein kinase (PKA) in a plethora of essential cellular processes. Here we show that yeast cells partially depleted of PKA activity (due to a tpk w mutation) and bearing a lesion in a Golgi-localized Ca2+ pump (Pmr1), arrest division with a small bud. The bud morphology of the arrested tpk1 w pmr1 mutant cells is characteristic of cells in S phase; however, the terminal phenotype of processes such as DNA replication and nuclear division suggests arrest at the G2/M boundary. This small bud, G2-arrest phenotype is similar to that of strains with a defect in cell wall biosynthesis (pkc1) or membrane biogenesis (och1); however, the biochemical defect may be different since the tpk1 w pmr1 double mutants retain viability. The growth defect of the tpk1 w pmr1 mutant can be alleviated by preventing the increase in cellular cAMP levels that is known to be associated with a decrease in PKA activity, or by supplementing the medium with millimolar amounts of Ca2+. Although the biochemical consequences of this increase in cAMP concentration are not known, the small-bud phenotype of the double mutant and the known protein processing defect of the pmr1 lesion suggest that the localization or function of some membrane component might be compromised and susceptible to perturbations in cellular cAMP levels. One candidate for such a protein is the cAMP-binding membrane ectoprotein recently described in yeast.
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  • 76
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 251 (1996), S. 556-564 
    ISSN: 1617-4623
    Schlagwort(e): cAMP ; Ca2+ ; Golgi apparatus ; Bud growth ; Saccharomyces cerevisiae
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Biochemical and physiological studies have implicated cAMP and cAMP-dependent protein kinase (PKA) in a plethora of essential cellular processes. Here we show that yeast cells partially depleted of PKA activity (due to atpk w mutation) and bearing a lesion in a Golgi-localized Ca2+ pump (Pmr1), arrest division with a small bud. The bud morphology of the arrestedtpk1 w pmr1 mutant cells is characteristic of cells in S phase; however, the terminal phenotype of processes such as DNA replication and nuclear division suggests arrest at the G2/M boundary. This small bud, G2-arrest phenotype is similar to that of strains with a defect in cell wall biosynthesis (pkc1) or membrane biogenesis (och1); however, the biochemical defect may be different since thetpk1 w pmr1 double mutants retain viability. The growth defect of thetpk1 w pmr1 mutant can be alleviated by preventing the increase in cellular cAMP levels that is known to be associated with a decrease in PKA activity, or by supplementing the medium with millimolar amounts of Ca2+. Although the biochemical consequences of this increase in cAMP concentration are not known, the small-bud phenotype of the double mutant and the known protein processing defect of thepmr1 lesion suggest that the localization or function of some membrane component might be compromised and susceptible to perturbations in cellular cAMP levels. One candidate for such a protein is the cAMP-binding membrane ectoprotein recently described in yeast.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 77
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 253 (1996), S. 173-181 
    ISSN: 1617-4623
    Schlagwort(e): Key words Illegitimate recombination ; Single-stranded plasmids ; Saccharomyces cerevisiae
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  We studied illegitimate recombination by transforming yeast with a single-stranded (ss) non-replicative plasmid. Plasmid pCW12, containing the ARG4gene, was used for transformation of yeast strains deleted for the ARG4, either in native (circular) form or after linearization within the vector sequence by the restriction enzyme ScaI. Both circular and linearized ss plasmids were shown to be much more efficient in illegitimate integration than their double-stranded (ds) counterparts and more than two-thirds of the transformants analysed contained multiple tandem integrations of the plasmid. Pulsed-field gel electrophoresis of genomic DNA revealed significant changes in the karyotype of some transformants. Plasmid DNA was frequently detected on more than one chromosome and on mitotically unstable, autonomously replicating elements. Our results show that the introduction of nonhomologous ss DNA into yeast cells can lead to different types of alterations in the yeast genome.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 78
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 253 (1996), S. 247-252 
    ISSN: 1617-4623
    Schlagwort(e): Key words Middle repetitive sequence ; Arabidopsis ; In situ hybridization ; Centromere ; Physical mapping
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  The middle repetitive fraction of the Arabidopsis genome has been relatively poorly characterized. We describe here a novel repetitive sequence cloned in the plasmid mi167, and present in ∼90 copies in the genome of Arabidopsis thaliana ecotype Columbia. Hybridization analysis to physically mapped YAC clones representing Arabidopsis chromosome 4 revealed four mi167-hybridizing loci, all clustered near the centromere. No other loci were detected on YAC clones covering chromosome 4. In situ hybridisation experiments to Arabidopsis chromosome spreads showed that mi167-hybridizing sequences are clustered at the centromeric heterochromatin of all five chromosomes; on two chromosomes the hybridization appeared to be localised on one arm. Additional mi167-hybridizing loci were detected, one of which was adjacent to a non-centromeric, heterochromatic region. This work supports the idea that the majority of middle repetitive DNA in the Arabidopsis genome is clustered. It also adds to our understanding of the organization of the centromere of Arabidopsis chromosome 4.
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    Standort Signatur Erwartet Verfügbarkeit
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  • 79
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 253 (1996), S. 81-88 
    ISSN: 1617-4623
    Schlagwort(e): Key words Rat ; DNA topoisomerase IIα ; Saccharomyces cerevisiae ; top2ts ; Complementation ; Leucine zipper
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  A series of yeast expression plasmids which comprise segments of the cDNA sequences encoding rat topo IIα have been constructed. The transcription of these constructs is under the control of the yeast GAL1 promoter. Galactose-dependent expression of the cloned rat topo IIα cDNA complemented a yeast top2 ts mutation, as well as a deletion mutation at the yeast TOP2 locus. Truncation of 12 N-terminal amino acids and/or 158 C-terminal amino acids of rat topo IIα had no effect on its ability functionally to substitute for top2 ts . Moreover, a cDNA construct with mutated putative leucine zipper domain (amino acids 993–1013) retained the complementation activity. These observations suggest that transformants capable of conditional topo IIα expression can be exploited as a useful model system for studies on the structure-function relationships of wild-type and mutated topo IIα, as well as the interplay of potential antitumor drugs with the enzyme.
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  • 80
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 253 (1996), S. 393-396 
    ISSN: 1617-4623
    Schlagwort(e): Key words Eukaryotic regulatory genes ; ADE2 gene ; Marked homologous recombination ; Gene-gene interference ; Saccharomyces cerevisiae
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  A novel method, which is rapid, reliable and quantitative, is presented for measuring the toxic effects on yeast cells of high dosage of any given gene. It is based on the possibility of monitoring the presence in cells of a plasmid carrying the ADE2 gene from Saccharomyces cerevisiae by direct observation of colonies, the construction of this particular plasmid being easily made by marked homologous recombination in yeast. Four yeast regulatory genes tested were found to result in various degrees of toxicity at high dosage. Possible implications of the measurement of gene toxicity for eukaryotic cell regulatory mechanisms and for the use of novel general approaches to gene selection, such as the gene-gene interference method, are discussed.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 81
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 252 (1996), S. 420-428 
    ISSN: 1617-4623
    Schlagwort(e): Saccharomyces cerevisiae ; Chitin synthases ; Septum
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Predicted protein sequences of fungal chitin synthases can be divided into a non-homologous N-terminal region and a C-terminal region that shows significant homology among the various synthases. We have explored the function of these domains by constructing a series of nested deletions, extending from either end, in theCHS1 andCHS2 genes ofSaccharomyces cerevisiae. In both cases, most or all of the sequences encoding the non-homologous N-terminal region (one-third of the protein for Chs1p and about one-fourth for Chs2p) could be excised, with little effect on the enzymatic activity in vitro of the corresponding synthase or on its function in vivo. However, further small deletions (20–25 amino acids) into the homologous region were deleterious to enzymatic activity and function, and often led to changes in the zymogenic character of the enzymes. Similarly, relatively small (about 75 amino acids) deletions from the C-terminus resulted in loss of enzymatic activity and function of both synthases. Thus, it appears that all the information necessary for membrane localization, enzymatic activity and function resides in the homologous regions of Chs1p and Chs2p, a situation that may also apply to other chitin synthases.
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  • 82
    ISSN: 1617-4623
    Schlagwort(e): 18S rRNA transcription ; Plant mitochondrial promoters ; Arabidopsis ; Pea ; Potato
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Transcription initiation sites for the mitochondrial 18S rRNA genes in the dicot plantsArabidopsis thaliana, potato and pea were identified by a combination of in vitro capping, primer extension and S-1 analyses. These promoters contain a nonanucleotide motif and an AT-rich sequence similar to many mRNA and tRNA promoters in dicot mitochondria. InArabidopsis and potato, active promoters are located within 120 nucleotides upstream of the 18S rRNA genes, as inOenothera. The nucleotide sequence in the corresponding region in pea mitochondria is well conserved, but is not used as promoter in this plant. Instead a novel promoter sequence is used that lies several hundred nucleotides upstream. These results show that rRNAs can be transcribed from the same promoter types as mRNAs and tRNAs in plant mitochondria. However, the sequence features presently attributed to plant mitochondrial promoters — the conserved nonanucleotide and the upstream AT-rich box — do not allow to deduce the presence of an active promoter from genomic sequence data alone.
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  • 83
    ISSN: 1617-4623
    Schlagwort(e): Saccharomyces cerevisiae ; Oxidative stress ; Pentose phosphate pathway ; Ribulose 5-phosphate epimerase
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract We have isolated several mutants ofSaccharomyces cerevisiae that are sensitive to oxidative stress in a screen for elevated sensitivity to hydrogen peroxide. Two of the sixteen complementation groups obtained correspond to structural genes encoding enzymes of the pentose phosphate pathway. Allelism of thepos10 mutation (POS forperoxidesensitivity) to thezwf1/met1 mutants in the structural gene for glucose 6-phosphate dehydrogenase was reported previously. The second mutation,pos18, was complemented by transformation with a yeast genomic library. The open reading frame of the isolated gene encodes 238 amino acids. No detectable ribulose 5-phosphate epimerase activity was found in thepos18 mutant, suggesting that the corresponding structural gene is affected in this mutant. For that reason the gene was renamedRPE1 (forribulose 5-phosphateepimerase).RPE1 was localized to chromosome X. The predicted protein has a molecular mass of 25 966 Daltons, a codon adaptation index (CAI) of 0.32, and an isoelectric point of 5.82. Database searches revealed 32 to 37% identity with ribulose 5-phosphate epimerases ofEscherichia coli, Rhodospirillum rubrum, Alcaligenes eutrophus andSolanum tuberosum. We have characterizedRPE1 by testing enzyme activities inrpe1 deletion mutants and in strains that overexpressRPE1, and compared the hydrogen peroxide sensitivity ofrpe1 mutants to that of other mutants in the pentose phosphate pathway. Interestingly, all mutants tested (glucose 6-phosphate dehydrogenase, gluconate 6-phosphate dehydrogenase, ribulose 5-phosphate epimerase, transketolase, transaldolase) are sensitive to hydrogen peroxide.
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  • 84
    ISSN: 1617-4623
    Schlagwort(e): DNA repair ; Nucleotide excision repair ; Saccharomyces cerevisiae
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Ultraviolet irradiation of DNA induces cyclobutane pyrimidine dimers (CPDs), 6-4′-(pyrimidine 2′-one) pyrimidines and pyrimidine hydrates. The dimer is the major photoproduct, and is specifically recognized by endonuclease V of phage T4. Pyrimidine hydrates represent a small fraction of the total photoproducts, and are substrates for endonuclease III ofEscherichia coli. We used these enzymes to follow the fate of their substrates in the mating type loci ofSaccharomyces cerevisiae. In a RAD strain, CPDs in the transcriptionally activeMATα locus are preferentially repaired relative to the inactiveHMLα locus, whilst repair of endonuclease III-sensitive sites is not preferential. Therad1, 2, 3 and4 mutants, which lack factors that are essential for the incision step of nucleotide excision repair (NER), repair neither CPDs nor endonuclease III-sensitive sites, clearly showing that these lesions are repaired by the NER pathway. Previously it had been shown that the products of theRAD7 andRAD16 genes are required for the NER of CPDs from theHMLα locus. We show that, in the same locus, these gene products are not needed for removal of endonuclease III-sensitive sites by the same mechanism. This indicates that the components required for NER differ depending on either the type of lesion encountered or on the specific location of the lesion within the genome.
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  • 85
    ISSN: 1617-4623
    Schlagwort(e): RCC1 ; Saccharomyces cerevisiae ; Serine/threonine protein kinase
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The yeast PRP20 protein is homologous to the RCC1 protein of higher eukaryotes and is required for mRNA export and maintenance of nuclear structure. RCC1/PRP20 act as guanine nucleotide exchange factors for the nuclear Ras-like Ran/GSP1 proteins. In a search forprp20-10 allele-specific high-copy-number suppressors, theKSP1 locus, encoding a serine/threonine protein kinase was isolated. Ksp1p is a nuclear protein that is not essential for vegetative growth of yeast. Inactivation of the kinase activity by a mutation affecting the catalytic center of the Ksp1p eliminated the suppressing activity. Based on the isolation of a protein kinase as a high-copy-number suppressor, the phosphorylation of Prp20p was examined. In vivo labeling experiments showed that Prp20p is a phosphoprotein; however, deletion of the KSP1 kinase did not affect Prp20p phosphorylation.
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  • 86
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 252 (1996), S. 552-562 
    ISSN: 1617-4623
    Schlagwort(e): Saccharomyces cerevisiae ; mRNA 3′ processing ; Poly(A) tail ; STS1 ; RNA15
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract In a search for proteins associated with Rna15p in processing the 3′ ends of messenger RNAs, we have looked for suppressors that correct, even partially, the thermosensitive growth defect of therna15-2 mutant. Mutations in a single locus that we namedSSM5, were able to suppress both the thermosensitivity of cell growth and the mRNA 3′ processing defect associated with therna15-2 mutation, but only slightly alleviated the thermosensitive growth defect of anrna14-1 mutant. Thessm5-1 mutant is sensitive to hydroxyurea at 37° C, a drug that inhibits DNA synthesis. By screening for complementation of the hydroxyurea-sensitive phenotype we cloned the corresponding wild-type gene and found that it corresponds to the essential geneSTS1 (also namedDBF8). Sts1p has an apparent molecular weight of 30 kDa and was confirmed to be a cytosolic protein by immunofluorescence analysis. Western blot analysis indicates that the thermosensitive mutant strainsrna15-2, rna14-1 andpap1-1 present a very low level of the Rna15p at 37° C. Thessm5-1 mutation restores the level of Rna15p in therna15-2 ssm5-1 double mutant. Use of the two-hybrid system suggests that Sts1p does not interact directly with Rna15p, but may be active as a homodimer. The present data suggest that Sts1p may play a role in the transport of Rna15p from the cytoplasm to the nucleus.
    Materialart: Digitale Medien
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  • 87
    ISSN: 1617-4623
    Schlagwort(e): Saccharomyces cerevisiae ; Mitochondrial pre-mRNA splicing ; NAM2 gene ; Leucyl tRNA synthetase ; RNA maturase
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract TheNAM2 gene ofSaccharomyces cerevisiae encodes the mitochondrial leucyl tRNA synthetase (mLRS), which is necessary for the excision of the fourth intron of the mitochondrialcytb gene (bI4) and the fourth intron of the mitochondrialcoxI gene (aI4), as well as for mitochondrial protein synthesis. Some dominant mutant alleles of the gene are able to suppress mutations that inactivate the bI4 maturase, which is essential for the excision of the introns aI4 and bI4. Here we report mutagenesis studies which focus on the splicing and suppressor functions of the protein. Small deletions in the C-terminal region of the protein preferentially reduce the splicing, but not the synthetase activity; and all the C-terminal deletions tested abolish the suppressor activity. Mutations which increase the volume of the residue at position 240 in the wild-type mLRS without introducing a charge, lead to a suppressor activity. The mutant 238C, which is located in the suppressor region, has a reduced synthetase activity and no detectable splicing activity. These data show that the splicing and suppressor functions are linked and that the suppressor activity of the mutant alleles results from a modification of the wild-type splicing activity.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 88
    ISSN: 1617-4623
    Schlagwort(e): Saccharomyces cerevisiae ; Mitochondria ; Integral membrane protein
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract SSM4 was isolated as a suppressor ofrna14-1, a mutant involved in nuclear mRNA maturation. In order to isolate genes interacting withSSM4, we have searched for mutants that are syntheticlethal in association with anSSM4 deletion. Among the mutants obtained, one, namedsls1-1, shows apet − phenotype. We have cloned and sequenced this gene. It encodes a protein with a calculated molecular mass of 73 kDa. This protein contains a mitochondrial targeting presequence but does not show homology with other known proteins. Deletion ofSLS1 does not affect cell viability on glucose but is lethal on a non-fermentable medium. The Sls1p protein does not appear to be involved in mitochondrial DNA replication, transcription, or in RNA splicing maturation or stability. We have also tagged this protein and localized it in mitochondria. Treatment with alkaline carbonate does not extract this protein from mitochondria, suggesting strongly that it is a mitochondrial integral membrane protein. Thus, theSLS1 gene, encodes a mitochondrial integral membrane protein and is paradoxically synlethal in association with a deletion of theSSM4 gene, which encodes an integral nuclear membrane protein.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 89
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 251 (1996), S. 261-266 
    ISSN: 1617-4623
    Schlagwort(e): Key words Biotin ; Arabidopsis ; Complementation ; bioA
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  The Arabidopsis thaliana biotin auxotroph bio1 was rendered prototrophic by transformation with a chimeric transgene containing the Escherichia coli bioA gene driven by a constitutive promoter. The bioA gene encodes the biotin biosynthetic enzyme 7,8-diaminopelargonic acid aminotransferase. Unlike the untransformed control plants, transgenic plants expressing the bacterial transgene synthesized biotin and grew to maturity without biotin-deficiency symptoms. These findings demonstrate that bio1/bio1 mutant plants are defective in the gene encoding 7,8-diaminopelargonic acid aminotransferase.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 90
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 253 (1996), S. 267-277 
    ISSN: 1617-4623
    Schlagwort(e): Key words KEULE ; Cytokinesis ; Arabidopsis ; Multinucleate cells ; Wall stubs
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  We present evidence to show that the KEULE gene of Arabidopsis is involved in cytokinesis. Mutant keule embryos have large multinucleate cells with gapped or incomplete cross walls, as well as cell wall stubs that are very similar to those observed upon caffeine inhibition of cytokinesis in plants. These defects are observed in all populations of dividing cells in the mutant, including calli, but less frequently in mature cells. Cell division appears to be slowed down, and the planes of cell division are often misoriented. In late embryos and seedlings, cross-wall formation usually appears complete, suggesting that the requirement for KEULE during cytokinesis is not absolute. Nonetheless, keule mutants die as seedlings with large polyploid cells. The bloated surface layer of keule seedlings does not uniformly behave like wild-type epidermis, and patches of this layer assume characteristics of the underlying ground tissue. The cytokinesis defect of keule mutants may influence aspects of cellular differentiation.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 91
    ISSN: 1617-4623
    Schlagwort(e): Key words DNA replication ; Meiosis ; Saccharomyces cerevisiae
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  In this report we study the regulation of premeiotic DNA synthesis in Saccharomyces cerevisiae. DNA replication was monitored by fluorescence-activated cell sorting analysis and by analyzing the pattern of expression of the DNA polymerase α-primase complex. Wild-type cells and cells lacking one of the two principal regulators of meiosis, Ime1 and Ime2, were compared. We show that premeiotic DNA synthesis does not occur in ime1Δ diploids, but does occur in ime2Δ diploids with an 8–9 h delay. At late meiotic times, ime2Δ diploids exhibit an additional round of DNA synthesis. Furthermore, we show that in wild-type cells the B-subunit of DNA polymerase α is phosphorylated during premeiotic DNA synthesis, a phenomenon that has previously been reported for the mitotic cell cycle. Moreover, the catalytic subunit and the B-subunit of DNA polymerase α are specifically degraded during spore formation. Phosphorylation of the B-subunit does not occur in ime1Δ diploids, but does occur in ime2Δ diploids with an 8–9 h delay. In addition, we show that Ime2 is not absolutely required for commitment to meiotic recombination, spindle formation and nuclear division, although it is required for spore formation.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 92
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 252 (1996), S. 510-517 
    ISSN: 1617-4623
    Schlagwort(e): Key words FK506 binding protein (FKBP) ; Immunophilins ; Tetratricopeptide repeat (TPR) ; Plant stress ; Arabidopsis
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  We have isolated clones of an Arabidopsis gene (ROF1, for rotamase FKBP) encoding a high molecular weight member of the FK506 binding protein (FKBP) family. The deduced amino acid sequence of ROF1 predicts a 551-amino acid, 62 kDa polypeptide which is 44% identical to human FKBP59 – a 59 kDa FKBP which binds to the 90 kDa heat shock protein and is associated with inactive steroid hormone receptors. ROF1 contains three FKBP12-like domains in the N-terminal portion of the protein (in contrast to two domains in mammalian FKBP59), an internal repeat structure associated with protein-protein interactions (tetratricopeptide repeats), and a putative calmodulin binding domain near the C-terminal region of the protein. No sequences associated with protein translocation out of the cytosol were found in ROF1. ROF1 mRNA was found at equivalent low levels in light-grown roots, stems, and flowers and at slightly higher levels in leaves. The abundance of ROF1 mRNA increased several-fold under stress conditions such as wounding or exposure to elevated NaCl levels.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 93
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 250 (1996), S. 162-168 
    ISSN: 1617-4623
    Schlagwort(e): Key words DNA repair ; Regulation ; Gene fusion ; DRE element ; Saccharomyces cerevisiae
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  The interstrand cross-link repair gene SNM1 of Saccharomyces cerevisiae was examined for regulation in response to DNA-damaging agents. Induction of SNM1-lacZ fusions was detected in response to nitrogen mustard, cis-platinum (II) diamine dichloride, UV light, and 8-methoxypsoralen +UVA, but not after heat-shock treatment or incubation with 2-dimethylaminoethylchloride, methylmethane sulfonate or 4-nitroquinoline-N-oxide. The promoter of SNM1 contains a 15 bp motif, which shows homology to the DRE2 box of the RAD2 promoter. Similar motifs have been found in promoter regions of other damage-inducible DNA repair genes. Deletion of this motif results in loss of inducibility of SNM1. Also, a putative negative upstream regulation sequence was found to be responsible for repression of constitutive transcription of SNM1. Surprisingly, no inducibility of SNM1 was found after treatment with DNA-damaging agents in strains without an intact DUN1 gene, while regulation seems unchanged in sad1 mutants.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 94
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 250 (1996), S. 162-168 
    ISSN: 1617-4623
    Schlagwort(e): DNA repair ; Regulation ; Gene fusion ; DRE element ; Saccharomyces cerevisiae
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The interstrand cross-link repair geneSNM1 ofSaccharomyces cerevisiae was examined for regulation in response to DNA-damaging agents. Induction ofSNM1-lacZ fusions was detected in response to nitrogen mustard, cis-platinum (II) diamine dichloride, UV light, and 8-methoxypsoralen + UVA, but not after heat-shock treatment or incubation with 2-dimethyl-aminoethylchloride, methylmethane sulfonate or 4-nitroquinoline-N-oxide. The promoter ofSNM1 contains a 15 bp motif, which shows homology to the DRE2 box of theRAD2 promoter. Similar motifs have been found in promoter regions of other damage-inducible DNA repair genes. Deletion of this motif results in loss of inducibility ofSNM1. Also, a putative negative up-stream regulation sequence was found to be responsible for repression of constitutive transcription ofSNM1. Surprisingly, no inducibility ofSNM1 was found after treatment with DNA-damaging agents in strains without an intactDUN1 gene, while regulation seems unchanged insad1 mutants.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 95
    ISSN: 1617-4623
    Schlagwort(e): Key words DNA repair ; Nucleotide excision repair ; Saccharomyces cerevisiae
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  Ultraviolet irradiation of DNA induces cyclobutane pyrimidine dimers (CPDs), 6-4′-(pyrimidine 2′-one) pyrimidines and pyrimidine hydrates. The dimer is the major photoproduct, and is specifically recognized by endonuclease V of phage T4. Pyrimidine hydrates represent a small fraction of the total photoproducts, and are substrates for endonuclease III of Escherichia coli. We used these enzymes to follow the fate of their substrates in the mating type loci of Saccharomyces cerevisiae. In a RAD strain, CPDs in the transcriptionally active MATα locus are preferentially repaired relative to the inactive HMLα locus, whilst repair of endonuclease III-sensitive sites is not preferential. The rad1, 2, 3 and 4 mutants, which lack factors that are essential for the incision step of nucleotide excision repair (NER), repair neither CPDs nor endonuclease III-sensitive sites, clearly showing that these lesions are repaired by the NER pathway. Previously it had been shown that the products of the RAD7 and RAD16 genes are required for the NER of CPDs from the HMLα locus. We show that, in the same locus, these gene products are not needed for removal of endonuclease III-sensitive sites by the same mechanism. This indicates that the components required for NER differ depending on either the type of lesion encountered or on the specific location of the lesion within the genome.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 96
    Digitale Medien
    Digitale Medien
    Springer
    Plant and soil 187 (1996), S. 91-95 
    ISSN: 1573-5036
    Schlagwort(e): Arabidopsis ; histogems ; root apical meristem ; root cap
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Arabidopsis thaliana roots have closed apical organization with three initial tiers. The dermatogen/calyptrogen tier consists of two parts-the central initials form the columella root cap, and the peripheral initial cells form the protoderm (epidermis) and the peripheral root cap. These peripheral initials divide in a sequence to form a root cap consisting of interconnected cones. the periblem initial tier forms the ground meristem (cortex). For the first week after germination the periblem consists of one layer of initial cells. The peripheral cells of the tier divide periclinally and then anticlinally (a T-division) to form the two-layered cortex (outer cortex and endodermis). After about one week, all the peripheral cells have divided periclinally forming two initials; the outermost produces the outer cortex while the inner initial produces the endodermis and middle cortex layer. The latter two cells arise via a periclinal division. During this time, other cells within the tier divide periclinally to form a two-layered tier. The plerome forms the cells of the procambium (vascular cylinder) by simple anticlinal divisions followed by longitudinal divisions to fill out the cell files of the vascular cylinder. A survey (27 dicot species in 17 families) of roots with closed apical organization revealed that there are three different types of root cap-concentric cylinders of cells (e.g.Linum), interconnecting cones (e.g.Arabidopsis) or overlapping arcs (e.g.Gossypium). H Lambers Section editor
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 97
    ISSN: 1573-5036
    Schlagwort(e): Arabidopsis ; cell axialization ; embryo ; root ; vascular tissues
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract In dicotyledonous plants, primordia of most seedling organs are laid down between fertilization and the formation of the heart-stage embryo. InArabidopsis embryogenesis, highly regular cell divisions and cell expansions facilitate the characterization of mutant development. We have taken a genetic approach to identify genes involved in organising the hypocotyl/root axis. The initiation of this axis is marked by oriented expansions and longitudinal division of cells in the lower tier of the early globular embryo. These cells go on to form a defined number of parallel cell files that constitute the hypocotyl and most of the radicle. Mutants impaired in the initiation or elaboration of the hypocotyl/root axis were selected by their seedling phenotype and subsequently analysed at embryonic stages. Several conclusions are suggested by the phenotypes of these mutants. First, hypocotyl/root axis formation can be genetically separated from other aspects of embryonic pattern formation. Second, initiation of the hypocotyl/root axis appears to be genetically distinct from post-embryonic root initiation. Third, four loci were identified that appear to contribute to the elaboration of the axial pattern. Finally, an anatomical inspection of one of the mutants, amenable to an analysis at post-embryonic stages, suggests a genetic link between basal pattern formation in the embryo and aspects of vascular differentiation in the adult plant. H Lambers Section editor
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 98
    Digitale Medien
    Digitale Medien
    Springer
    Plant growth regulation 18 (1996), S. 71-77 
    ISSN: 1573-5087
    Schlagwort(e): Arabidopsis ; ethylene ; ethylene binding protein ; signal transduction
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract A review of work carried out on ethylene binding in higher plants is presented. The use of radio-labelled displacement assays has identified specific 14C-ethylene binding in all tissues so far studied. virtually all higher plants studied contain at least two classes of ethylene binding site, one of which fully associates and dissociates in about 2 h and a class of sites that takes up to 20 h to become fully saturated. Although the types of site differ in their rate constants of association they have similar and high affinities for ethylene. A series of Arabidopsis thaliana mutants shown to vary in sensitivity to ethylene have been analysed for 14C-ethylene binding. One mutant, eti 5, which was shown to be unaffected by ethylene concentrations of up to 10,000 μL L−1 was also shown to exhibit reduced binding. In vivo and in vitro studies on pea have shown that ethylene binding can be detected in this tissue. In vitro studies have shown that both membrane and cytosolic fractions contain measurable amounts of ethylene binding. Interestingly, cytosolic ethylene binding consisted only of the fast associating/dissociating type. Developing cotyledons of Phaseolus vulgaris contain a higher concentration of ethylene binding sites that other tissues and only contain the slow dissociating component. These facets have allowed the purification of ethylene binding protein(s) (EBP) from this tissue. The proteins which bind ethylene can be resolved into two bands of 26 and 28 kDa on semi-denaturing PAGE and the proteins appear to be single entities on a 2-D gels. Data will be presented which indicate a possible role for heterotrimetric G-proteins in the early stages of the ethylene signal transduction pathway.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 99
    ISSN: 1573-6857
    Schlagwort(e): Arabidopsis ; heterochromatin ; retroviral element ; satellite DNA ; transposon
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract An analysis of Arabidopsis thaliana heterochromatic regions revealed that genomic sequences immediately flanking the major 180 bp satellite are essentially made of middle repetitive sequences and that most of these sequences correspond to defective Athila retroelements. Using YAC and λ clones, we evaluated the distribution of Athila elements in the Arabidopsis genome and showed that, despite the presence of numerous euchromatic copies, these elements are especially concentrated in or near heterochromatic regions. Sequencing of the various DNA transitions between satellite and Athila repeats provides strong evidence that most of the heterochromatic elements retrotransposed directly into 180 bp satellite clusters.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 100
    Digitale Medien
    Digitale Medien
    Springer
    Journal of industrial microbiology and biotechnology 16 (1996), S. 117-123 
    ISSN: 1476-5535
    Schlagwort(e): baker's yeast ; Saccharomyces cerevisiae ; fed-batch cultivation ; ethanol sensor
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Werkstoffwissenschaften, Fertigungsverfahren, Fertigung
    Notizen: Abstract Growth kinetics ofSaccharomyces cerevisiae in glucose syrup from cassava starch and sugarcane molasses were studied using batch and fed-batch cultivation. The optimum temperature and pH required for growth were 30°C and pH 5.5, respectively. In batch culture the productivity and overall cell yield were 0.31 g L−1 h−1 and 0.23 g cells g−1 sugar, respectively, on glucose syrup and 0.22 g L−1 h−1 and 0.18 g cells g−1 sugar, respectively, on molasses. In fed-batch cultivation, a productivity of 3.12 g L−1 h−1 and an overall cell yield of 0.52 g cells g−1 sugar in glucose syrup cultivation and a productivity of 2.33 g L−1 h−1 and an overall cell yield of 0.46 g cells g−1 sugar were achieved in molasses cultivation by controlling the reducing sugar concentration at its optimum level obtained from the fermentation model. By using an on-line ethanol sensor combined with a porous Teflon® tubing method in automating the feeding of substrate in the fed-batch culture, a productivity of 2.15 g L−1 h−1 with a yield of 0.47 g cells g−1 sugar was achieved using glucose syrup as substrate when ethanol concentration was kept at a constant level by automatic control.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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