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  • wheat  (57)
  • Saccharomyces cerevisiae  (43)
  • Yeast  (35)
  • Springer  (133)
  • American Institute of Physics
  • Blackwell Publishing Ltd
  • 2020-2023
  • 1990-1994  (133)
  • 1980-1984
  • 1925-1929
  • 1990  (133)
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  • Springer  (133)
  • American Institute of Physics
  • Blackwell Publishing Ltd
  • Wiley-Blackwell  (14)
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  • 2020-2023
  • 1990-1994  (133)
  • 1980-1984
  • 1925-1929
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  • 1
    ISSN: 1476-5535
    Keywords: Lignocellulosic waste ; Yeast ; Ethanol production ; Optimization study
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Hardwood hemicellulose hydrolysate has been utilized as a substrate for ethanol production. Among the three different yeasts tested, the best performances have been obtained, in decreasing order, usingPachysolen tannophilus, Candida shehatae andPichia stipitis. Several pretreatments of this raw material have been studied to improve ethanol yields; in one such pretreatment a strain ofP. tannophilus produced ethanol with a yield of 0.29 gethanol/gsugars (gP/gS); which is only 15% less than the values observed with synthetic media. Neither aeration nor acetone addition improved the fermentation of this substrate; in fact, only a marked stimulation of biomass growth has been observed at the expense of both ethanol and xylitol production.
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  • 2
    ISSN: 1432-0983
    Keywords: 2-oxoglutarate dehydrogenase ; Saccharomyces cerevisiae ; rad52-mediated chromosome loss
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Ogd1 mutants of Saccharomyces cerevisiae are deficient in mitochondrial 2-oxoglutarate dehydrogenase activity; they cannot grow on glycerol and produce an increased amount of organic acids during growth on glucose as substrate. Using gamma ray-induced rad52-mediated chromosome loss the ogd1 mutation can be assigned to chromosome IX. Tetrad analysis of crosses between ogd1 and other markers on chromosome IX revealed that the OGD1 gene maps on the left arm of this chromosome 1.9 cM from his5.
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  • 3
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Orotate phosphoribosyl transferase ; Nucleotide sequence-5-phosphoribosyl 1-pyrophosphate (5PRPP)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Orotate phosphoribosyl transferase (OPRTase) catalyses the transformation of orotate to OMP in the pyrimidine pathway. In the yeast Saccharomyces cerevisiae, the URA5 gene is known to encode this enzyme activity. In this paper we present the cloning and sequencing of a yeast gene, named URA10, encoding a second OPRTase enzyme. Comparison of the predicted amino acid sequences between URA5 and URA10 genes shows more than 75% similarity. These sequences have also been compared to those of Escherichia coli, Podospora anserina, Sordaria macrospora and Dictyostelium discoideum. Remarkable similarities in the primary structure of these proteins have been found. Gene disruption experiments revealed that URA10 gene expression is responsible for the leaky phenotype of a ura5 mutant. Assays of OPRTase activity in extracts from ura5 and ura10 mutants indicate that the URA10 product contributes only 20% of the total activity found in wild type cells.
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  • 4
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    Springer
    Current genetics 17 (1990), S. 269-273 
    ISSN: 1432-0983
    Keywords: Yeast ; Actin ; Intron ; Antisense RNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Using Northern blot analysis we have detected an approximately 840 nucleotide-long RNA which is complementary to the 5′ leader sequence and the first ten nucleotides of the coding sequence of the yeast actin (ACT1) messenger RNA. We have determined two transcription start sites for this actin antisense RNA (ASR1), both within the ACT1 intron, at about 80 and 90 nucleotides downstream from the 5′ splice site. Analysis of a cDNA clone showed that this RNA species overlaps the entire trailer sequence and approximately 20 nucleotides of the coding sequence of the nearby yeast YPT1 gene.
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  • 5
    ISSN: 1432-0983
    Keywords: Yeast ; Mutants ; Cytochrome ; Mitochondria ; Oxidative phosphorylation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary This paper reports studies of bioenergetic modifications in a TTR1 single-nuclear mutant, isolated as resistant to triethyltin, an inhibitor of mitochondrial ATPase, and effective in cAMP-dependent protein phosphorylation. This mutant appears to have lost the wildtype cell ability to respond to a decrease of oxygen concentration in the growth medium by a decrease of cytochrome concentration in the cell. ATP synthesis rate in mutant cells in both the prestationary and stationary phase of growth appeared increased in comparison to wild-type cells, as too was respiration rate. A comparative study of mitochondria extracted from wild-type and from TTR1 mutant cells showed an increase in respiration rate, an increase in ATP synthesis rate, and an increase in TPP+ uptake in mutant mitochondria. The specific ATPase activity, as well as its sensitivity to TET, appears to be similar for mitochondria extracted from both strains. It was proposed that the modification of mitochondrial biogenesis in the TTR1 mutant may be due to a response of the cell to an increase in ATP hydrolysis caused by the mutation. It is also possible that the modification in cAMP-dependent protein kinase regulation which appeared to occur in this mutant affects protein(s) involved in mitochondrial biogenesis.
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  • 6
    ISSN: 1432-0983
    Keywords: Yeast ; Chromosome mapping ; Acidic ribosomal proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary DNA probes from the genes encoding the acidic ribosomal proteins L44, L44′ and L45, as well as from reporter genes for chromosomes IV, VII, XII and XV, have been hybridised to Southern blots of Saccharomyces cerevisiae DNA resolved by pulsed field gel electrophoresis. The protein L44′ and protein L45 genes have been found to hybridise to chromosome IV, identified by the CAT1 gene probe, while the protein L44 probe hybridises with a band containing chromosomes VII and XV, identified by the ATPase 1 and HIS3 genes respectively.
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  • 7
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Mutants ; Farnesyl diphosphate synthetase ; Ergosterol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two yeast mutant strains auxotrophic for ergosterol and blocked in farnesyl diphosphate synthetase (EC 2.5.1.1) were isolated. Genetic analysis has shown that these mutant strains carry additional mutations in the ergosterol pathway besides erg20-1 and erg20-2 which affect FPP synthetase. The novel feature of these mutants is their ability to excrete prenyl alcohols (farnesol and geraniol). As geraniol is toxic for yeast cells, the above leaky mutations in FPP synthetase have to be associated with others in the sterol pathway, in order to slow down geraniol synthesis.
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  • 8
    ISSN: 1432-0983
    Keywords: Glucose oxidase ; Aspergillus ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We report the cloning of the Aspergillus niger glucose oxidase gene and its use to elevate glucose oxidase productivity in A. niger by increasing the gene dosage. In addition, the gene has been introduced into A. nidulans where it provides the novel capacity to produce glucose oxidase. A plasmid, in which DNA encoding the mature form of glucose oxidase was preceded by a Saccharomyces cerevisiae secretion signal, effected high-level production of extracellular glucose oxidase in this yeast.
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  • 9
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    Current genetics 18 (1990), S. 401-403 
    ISSN: 1432-0983
    Keywords: Baking yeast ; Saccharomyces cerevisiae ; Dough leavening ; Benomyl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary To investigate the leavening ability of yeast in dough, chromosome loss was induced by benomyl treatment in YOY1037, a diploid between a baking strain and a laboratory strain, and its effect on the leavening ability was studied. When benomyl-treated cells were spread on plates with a dye indicator for ploidy, about 20% of the visible colonies were stained dark blue or dark purple; the rest stained pale blue, similar to the diploid YOY1037. Strains showing the MATα phenotype, and non-galactose fermenting strains, apparently having lost particular chromosomes, were observed only in those with darkcoloured colonies. Strains with dark-coloured colonies showed a wider range of leavening ability than did those with pale-coloured colonies.
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  • 10
    ISSN: 1432-0983
    Keywords: Xylitol dehydrogenase gene ; Pichia stipitis ; Saccharomyces cerevisiae ; Xylose utilization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A P. stipitis cDNA library in λgt11 was screened using antisera against P. stipitis xylose reductase and xylitol dehydrogenase, respectively. The resulting cDNA clones served as probes for screening a P. stipitis genomic library. The genomic XYL2 gene was isolated and the nucleotide sequence of the 1089 bp structural gene, and of adjacent non-coding regions, was determined. The XYL2 open-reading frame codes for a protein of 363 amino acids with a predicted molecular mass of 38.5 kDa. The XYL2 gene is actively expressed in S. cerevisiae transformants. S. cerevisiae cells transformed with a plasmid, pRD1, containing both the xylose reductase gene (XYL1) and the xylitol dehydrogenase gene (XYL2), were able to grow on xylose as a sole carbon source. In contrast to aerobic glucose metabolism, S. cerevisiae XYL1-XYL2 transformants utilize xylose almost entirely oxidatively.
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  • 11
    ISSN: 1432-0983
    Keywords: Yeast ; Ribosomal protein gene ; Sequence analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two genes encoding ribosomal protein YL41 were cloned from Saccharomyces cerevisiae chromosomal DNA. Both genes contain an uniterrupted region of only 75 nucleotides coding for a protein of 3.3 kD. Within the coding regions the nucleotide sequences are virtually identical, whereas in both the 5′-and 3′-flanking regions the two genes differ significantly from each other. The deduced protein shows an arginine and lysine content of 68 percent, i.e., 17 out of 25 residues, and the basic residues are evenly distributed over the molecule. When compared to the ribosomal protein sequences currently available no counterpart to YL41 could be found in prokaryotes and it seems likely that YL41 is a eukaryotespecific ribosomal protein.
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  • 12
    ISSN: 1432-0983
    Keywords: Yeast ; Trans-acting Factor ; RAP1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We show by deletion mutagenesis, followed by in vivo and in vitro analysis, that the binding of a protein factor to the upstream activation sequence (USA) of the Saccharomyces cerevisiae glycolytic gene PYK, encoding pyruvate kinase, is required for efficient transcription of the corresponding coding region. In addition, gel electrophoretic mobility shift and DNase I protection studies, involving yeast gene products expressed in E. coli, suggest that this trans-acting DNA-binding protein is encoding by the RAP1 gene. The identification of RAP1 binding sites located within the UAS element of the yeast PYK, PGK (phosphoglycerate kinase) and ENO1 (enolase) genes, and in the 5′-upstream region of the ADHI (alcohol dehydrogenase) gene, suggests that a mechanism of coordinate gene expression involving several of the glycolytic genes may exist in yeast.
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  • 13
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    Current genetics 18 (1990), S. 485-491 
    ISSN: 1432-0983
    Keywords: Yeast ; Ty elements ; Virus like particles
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Electron microscopic analysis of thin sections of Saccharomyces cerevisiae NCYC74 has revealed the presence of many clumped cytoplasmic particles that morphologically resemble Ty element virus-like particles (VLPs). Accumulation of Ty VLPs has only previously been observed in S. cerevisiae strains that over-express a cloned Ty element. The particles in NCYC74 co-purify with Ty RNA, Ty-specific antigens and a reverse transcriptase activity. Furthermore, they appear to be recognised by antibodies to Ty VLPs during indirect immunofluorescence experiments. These observations provide compelling evidence that the cytoplasmic particle in NCYC74 are indeed Ty VLPs.
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  • 14
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    Current genetics 18 (1990), S. 29-39 
    ISSN: 1432-0983
    Keywords: Recombination ; DNA repair ; UV irradiation ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A novel synthetic locus is described that provides a simple assay system for characterizing mitotic recombinants. The locus consists of the TRP1 and HIS3 genes inserted into chromosome III of S. cerevisiae between the CRY1 and MAT loci. Defined trp1 and his3 alleles have been generated that allow the selection of interchromosomal recombinants in this interval. Trp+ or His+ recombinants can be divided into several classes based on coupling of the other alleles in the interval. The tight linkage of the CRY1 and MAT loci, combined with the drug resistance and cell type phenotypes that they respectively control, facilitates the classification of the recombinants without resorting to tetrad dissection. We present the distribution of spontaneous recombinants among the classes defined by this analysis. The data suggest that the recombination intermediate can have regions of symmetric strand exchange and that co-conversion tracts can extend over 1–3 kb. Continuous conversion tracts are favored over discontinuous tracts. The distribution among the classes defined by this analysis is altered in recombinants induced by UV irradiation.
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  • 15
    ISSN: 1432-0983
    Keywords: Yeast ; Mitochondrial RNA splicing ; Nuclear pet - mutant ; Group I introns
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have studied the role of the product of the nuclear gene PET157 in mitochondrial pre-mRNA splicing. Cytoduction experiments show that a mitochondrial genome deleted for the three introns bI3, aI5 and aI6 is able to suppress the pet157-1 mutation: the strain recovers respiratory competency indicating that the product of the PET157 gene is only required for mitochondrial premRNA splicing. Characterization of the high molecular weight pre-mRNAs which accumulate in the pet157 mutant demonstrate that the product of the PET157 gene is required for the excision of two group I introns bI3 and aI6 (corresponding to aI5β) located in the cob-box and coxI genes respectively. Furthermore, the pet157 mutant strain accumulates the bI3 maturase in the form of a polypeptide of 50K (p50) previously observed in mitochondrial mutants defective in the excision of bI3. We have shown by restriction analysis and allelism tests that the pet157-1 mutation is allelic to the nuclear mrs1 mutation, previously described as specifically blocking the excision of bI3. Finally, revertants obtained by the deletion of bI3 or aI6 from the mitochondrial DNA were isolated from the MRS1 disrupted allele, confirming the involvment of the product of the MRS1/PET157 gene in the excision of the two introns bI3 and aI6.
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  • 16
    ISSN: 1432-0983
    Keywords: Mutagen hyper-resistance ; Nitrogen mustard ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A screening of haploid yeast strains for enhanced resistance to nitrogen mustard (HN2) yielded a recessive mutant allele, hnm1, that conferred hyper-resistance (HYR) to HN2. Diploids, homo- or heterozygous for the HNM1 locus, exhibit normal wild-type like resistance while homozygosity for hnm1 leads to the phenotype HYR to HN2. The hnm1 mutation could be found in yeast strains proficient or deficient in different DNA repair systems. In these mostly HN2-sensitive haploid repair-deficient mutants, hnm1 acted as a partial suppressor of HN2 sensitivity. All isolated recessive mutations conferring hyper-resistance belonged to a single complementations group. The HYR to HN2 phenotype was maximally expressed in growing cells and was associated with reduced mutability by HN2. HNM1 most probably controls uptake of HN2 which would be impaired in the hnm1 mutants.
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  • 17
    ISSN: 1432-0983
    Keywords: S. cerevisiae ; Yeast ; Cadmium resistance ; CAD2 gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A cadmium-resistant strain, X3382-3A, which is able to grow in a medium containing 0.2 mM cadmium sulfate, was picked out from our laboratory stock strains of Saccharomyces cerevisiae. The cadmium resistance of this strain is controlled by a single dominant nuclear gene, denoted as CAD2. The locus of CAD2 was mapped by gene linkage to a site 15.5 centimorgans to the right of the his7 locus on the right arm of chromosome II. The cadmium resistance of the strain carrying CAD2 was evaluated for its properties of cadmium uptake, cadmium distribution and cadmium-metallothionein formation, in comparison with those of some other strains. The results suggest that the novel type of cadmium resistance controlled by CAD2 does not involve production of a cadmiumm-metallothionein.
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  • 18
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; G418 resistance ; Gene cartridges ; Heterologous Gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Coding sequence cartridges for aminoglycoside phosphotransferase (APT) were isolated from bacterial transposon Tn903. When incorporated into a heterologous gene construction utilising the PGK1 promoter and terminator, the heterologous APT gene provided a G418-resistance determinant that functioned efficiently as a dominant marker for yeast in both multiple- and single-copy. Transformant colonies on selective medium appeared rapidly, within 36–48 h, and growth rate of the transformed cells was normal. A simple and highly sensitive radiolabelling assay for APT enzyme activity was developed for use with crude cell protein extracts. Enzyme activity units were equated to the amount of APT protein present in the cells, and the APT protein was shown to be stable in yeast. Heterologous APT expression was 130-fold reduced compared with homologous PGK1. This resulted from an estimated two-fold decrease in mRNA level and a 65-fold decrease in translation efficiency. The latter was unaffected by AUG sequence context change, but corresponded with a high frequency of minor codons in the APT-coding sequence. APT can be used as a semi-quantitative reporter of gene expression, whose useful features are in vivo detection via the G418-resistance phenotype and powerful cell-free assay.
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  • 19
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Episomal plasmid ; Copy number control ; Plasmid maintenance ; Glycolytic enzyme levels
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary This study demonstrates how varying the promoter strength of an essential gene on a yeast 2μORI-STB YEp multicopy vector can influence vector copy levels. A phosphoglycerate kinase gene (PGK) on this plasmid was made essential for fermentative growth by transformation into a pgk - yeast strain. When in these PGK- transformants the requirement for PGK expression was the sole selective criterion for plasmid maintenance, PGK promoter activity was inversely related to vector copy levels. Plasmids with an efficiently-transcribed PGK gene were maintained at approximately one copy per cell, whereas those lacking the UAS that normally directs high basal PGK transcription levels were present at up to 10–15 copies. All cultures of these PGK+ transformants contained only a low proportion of pgk - cells. Since mitotic loss of the plasmid arrests growth through loss of a functional PGK allele, PGK confers high stability to the YEp vector in such a pgk - genetic background. In this system YEp vector levels are probably influenced by PGK transcription because high expression of PGK is needed in rapid fermentative growth. Remarkably, low plasmid PGK promoter activity caused PGK mRNA levels slightly higher than those found in yeast with normal PGK regulation. A higher plasmid copy number is therefore not the only factor counteracting the effects of low PGK transcription, and it is possible that PGK mRNA becomes more stable in response to inefficient PGK transcription.
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  • 20
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    Current genetics 17 (1990), S. 179-183 
    ISSN: 1432-0983
    Keywords: Heme ; Cytochromes ; Regulation ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Simultaneous effects of mutations in the transcriptional regulatory genes, HAP1, HAP2 and HAP3, on all respiratory cytochromes of Saccharomyces cerevisiae were determined. Cytochrome behavior in hap mutants and in cyc4 and rhm1 mutants, altered in regulation of 5-aminolevulinate synthase, was compared. Although hap mutants were isolated as trans-acting, transcriptional regulators of the CYC1 (iso-1-cytochrome c) gene, each mutant exhibits partial deficiencies in all cytochrome types. In hap2 and hap3 strains all cytochromes were decreased proportionally to about 40–50% of wild type values. In contrast, hap1 caused a decrease in all cytochromes and an accumulation of a pigment, probably Zn porphyrin. Apparently apocytochrome and heme biosynthesis retain coordination in hap2 and hap3, but not in hap1, mutants. Unlike cyc4 and rhm1 mutants, hap mutants do not exhibit 5-aminolevulinate-dependent restoration of cytochromes. The hap1 mutant grew at nearnormal rates on glycerol, whereas hap2 and hap3 mutants grew very slowly. The frequency of [rho-] was high (16–18%) in hap2 and hap3 strains. Results are consistent with generalized control of mitochondrial replication directed by the HAP1-HAP2 system and heme-directed control of formation of all apocytochromes mediated by HAP1. Neither system exerts all-or-nothing control.
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  • 21
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    Current genetics 17 (1990), S. 275-280 
    ISSN: 1432-0983
    Keywords: Yeast ; DNA replication ; Effect on mutation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Incubation in YPD medium under permissive conditions when DNA replication is going on, strongly stimulates the induction of cdc+ colonies of UV-irradiated cells of yeast strains HB23 (cdc8-1/cdc8-3), HB26 (cdc8-3/cdc8-3) and HB7 (cdc8-1/cdc8-1). Inhibition of DNA replication by hydroxyurea, araCMP, cycloheximide or caffeine or else by incubation in phosphate buffer pH 7.0, abolishes this stimulation. Thus the replication of DNA is strongly correlated with the high induction of cdc+ colonies by UV irradiation. It is postulated that these UV-induced cdc+ colonies arise as the result infidelity in DNA replication.
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  • 22
    ISSN: 1432-0983
    Keywords: Yarrowia lipolytica ; 7SL RNA ; Essential genes ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Cells containing a deletion of either the SCR1 or SCR2 genes, which code for the 7SL RNA component of the signal recognition particle (SRP) homologue, were found to be viable. Two independent approaches demonstrated that cells containing deletions of both genes were inviale. Therefore, Yarrowia lipolytica contains two (and only two) functional 7SL RNA genes.
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  • 23
    ISSN: 1432-0983
    Keywords: Mitotic recombination ; Hyper-recombination ; RAD50 ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mutations in the RAD50 gene of Saccharomyces cerevisiae have been shown to reduce double strand break repair, meiotic recombination, and radiation-inducible mitotic recombination. Several different point mutations (including ochre and amber alleles) have been previously examined for effects on spontaneous mitotic recombination and did not reduce the frequency of recombination. Instead, the rad50 mutations conferred a moderate hyper-rec phenotype. This paper examines a deletion/interruption allele of RAD50 that removes 998 of 1312 amino acids and adds 1.1 kb of foreign DNA. The results clearly indicate that spontaneous mitotic recombination can occur in the absence of RAD50; in fact, the frequency of recombination is elevated over the wild-type cell. One possible interpretation of these observations is that the initiating lesion in spontaneous recombination events in mitosis might not be a double strand break.
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  • 24
    ISSN: 1432-0983
    Keywords: Flow cytometry ; Rhodamine 123 ; Respiratory chain ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The cationic lipophilic dye Rhodamine 123 (Rh123) is selectively enriched in mitochondria in a membrane potential-dependent manner. Application of drugs which interfere with the electron flow of the respiratory chain lead to a severe reduction of mitochondrial dye uptake. In this communication we show that the same effect is observed after Rh123-staining of respiratory-deficient yeast mutants. Based on this observation we used flow cytometry to discriminate respiratory-compentent and respiratory-deficient yeast cells. Combined with a cell sorter we were able to selectively enrich respiring and non-respiring yeast cells, repectively, from a mixture of cells.
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  • 25
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Sporulation ; Inessential genes ; Genome organization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The SPR6 gene of Saccharomyces cerevisiae encodes a moderately abundant RNA that is present at high levels only during sporulation. The gene contains a long open reading frame that could encode a hydrophilic protein approximately 21 kDa in size. This protein is probably produced by the yeast, because the lacZ gene of Escherichia coli is expressed during sporulation when fused to SPR6 in the expected reading frame. SPR6 is inessential for sporulation; mutants that lack SPR6 activity sporulate normally and produce viable ascospores. Nonetheless, the SPR6 gene encodes a function that is relevant to sporulating cells; the wild-type allele can enhance sporulation in strains that are defective for several SPR functions. SPR6 is located on chromosome V, 14.4 centimorgans centromere-distal to MET6.
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  • 26
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Nucleo-mitochondrial interactions ; Mitochondrial status ; Lycorine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In a previous paper we have shown that the alkaloid lycorine inhibits growth of rho +, mit - and rho -, strains of Saccharomyces cerevisiae, whereas strains devoid of mitochondrial DNA (rho o) are resistant to more than 200 μg/ml of the alkaloid. In this report we show that hypersuppressive petites are almost as resistant as rho o mutants, whereas isogenic rho - petites, which have retained tained longer segments of the genome, are sensitive to the drug.
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  • 27
    ISSN: 1432-0983
    Keywords: Yeast ; QH2: cytochrome c oxidoreductase ; Mitochondrial biogenesis ; Transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In Saccharmmyces cerevisiae, the COR2 gene codes for the 40 kDa subunit II of the QH2: cytochrome c oxidoreductase, an enzyme of the mitochondrial respiratory chain. Regions in the 5′ flank of this gene important for regulated expression were identified by assaying β-galactosidase activities in cells carrying different COR2-lacZ fusion genes. Sequences downstream of position-201 relative to the translational initiation codon are sufficient to confer regulation by carbon source, whereas sequences downstream of position-153 do not give rise to significant expression. A binding site for the abundant general transcription factor GFI is present in the region between-201 and-153 just upstream from sequences which resemble the consensus DNA recognition sequence of the regulatory protein complex HAP2/HAP3: 5′-TNATTGGT-3′. By quantitating RNA levels and assaying β-galactosidase activities we show that synthesis of COR2, which is not a hemoprotein, is regulated by HAP1, HAP2/HAP3 and heme.
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  • 28
    ISSN: 1432-0983
    Keywords: Yeast ; Mutant ; Triethyltin chloride ; Protein phosphorylation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Three mutants of Saccharomyces cerevisiae resistant to triethyltin (an inhibitor of mitochondrial ATPase) on non-fermentative media, and non-resistant to this drug on fermentative media, were isolated and named TTR1, TTR2 and TTR3. Apart from triethyltin resistance, these mutants show the following common characteristics: (1) Increased intracellular cytochrome c concentration. (2) Increased respiration rate. (3) Decreased growth yield. (4) Increased growth sensitivity to several drugs inhibiting oxidative phosphorylation: namely, CCCP (permeabilizing inner mitochondrial membrane to protons), valinomycin (permeabilizing inner mitochondrial membrane to potassium) and oligomycin (inhibitor of mitochondrial ATPase). (5) Increased sensitivity to carbon source starvation. For each mutant, these characteristics appeared to be due to a single pleiotropic nuclear mutation. Mutation TTR1 causes additional phenotypic characteristics which do not appear in mutants TTR2 and TTR3: (1) Pinkish coloration of colonies which is more pronounced after a long growth period. (2) Inability of the cells to store glycogen. (3) Growth defect of the cells on a galactose-containing medium. (4) Inability of a diploid homozygote mutant strain to sporulate. All these phenotypic characteristics have already been described in yeast mutants deregulated in cAMP-dependant protein phosphorylation. Crossing of a strain bearing the TTR1 mutation with a strain mutated in the adenylate cyclase structural gene suggested that the TTR1 phenotype is due to a modification in regulation of cAPK by cAMP, making cell multiplication possible without intracellular cAMP.
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  • 29
    ISSN: 1432-0983
    Keywords: Schizosaccharomyces pombe ; Saccharomyces cerevisiae ; CaMV 35S promoter ; CaMV 35S terminator ; Heterologous expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Complementation of fission yeast mutants by plant genomic libraries could be a promising method for the isolation of novel plant genes. One important prerequisite is the functioning of plant promoters and terminators in Schizosaccharomyces pombe and Saccharomyces cerevisiae. Therefore, we studied the expression of the bacterial β-glucuronidase (GUS) reporter gene under the control of the Cauliflower Mosaic Virus (CaMV) 35S promoter and 35S terminator. We show here that S. pombe initiates transcription at exactly the same start site as was reported for tobacco. The 35S CaMV terminator is appropriately recognized leading to a polyadenylated mRNA of the same size as obtained in plant cells transformed with the same construct. Furthermore, the GUS-mRNA is translated into fully functional GUS protein, as determined by an enzymatic assay. Interestingly, expression of the 35S promoter in the budding yeast S. cerevisiae was found to be only moderate and about hundredfold lower than in S. pombe. To investigate whether different transcript stabilities are responsible for this enormous expression difference in the two yeasts, the 35S promoter was substituted by the ADH (alcohol dehydrogenase) promoter from fission yeast. In contrast to the differential expression pattern of the 35S promoter, the ADH promoter resulted in equally high expression rates in both fission and budding yeast, comparable to the 35S promoter in S. pombe. Since the copy number of the 35S-GUS constructs differs only by a factor of two in the two yeasts, it appears that differential recognition of the 35S promoter is responsible for the different transcription rates.
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  • 30
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    Current genetics 17 (1990), S. 493-497 
    ISSN: 1432-0983
    Keywords: Mitochondria ; Yeast ; Petites
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A pleiotropic, respiration-deficient mutant was isolated from the petite negative yeast Pachysolen tannophilus after UV mutagenesis. The mutant is unable to utilize xylose, arabinose, galactose or glycerol, and shows no detectable respiration when grown on glucose. Cytochrome c oxidase, xylose reductase and xylitol dehydrogenase activities are lacking. Mitochondrial ultrastructre is altered. The results support the hypothesis that functioning mitochondria are necessary for xylose utilization in this organism.
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  • 31
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Mitochondria ; Intron-encoded proteins ; Recombination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The respiratory competency of a yeast strain devoid of mitchondrial introns is quite normal. However, it may be asked whether intron-encoded proteins participate in metabolisms other than those of mitochondrial introns. Using strains without mitochondrial introns we have answered two questions. The first was: does the absence of intron-encoded proteins abolsh mitochondrial recombination? The second was: do mitochondrial introns and intron-encoded proteins play a part in mitochondrial DNA rearrangements induced by ethidium bromide (rho- production)? We have shown that the introns and intron-encoded proteins are not essential essential components of either phenomenon.
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  • 32
    ISSN: 1432-0983
    Keywords: Yeast ; Ty2 ; Protein/DNA binding ; Transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A series of transposable element-induced mutations at the HIS4 locus in Saccharomyces cerevisiae have been attributed to the transposition of a Ty element into the 5′ regulatory region of this gene. Various Ty-containing His+ revertants have been isolated and the HIS4/Ty junction region sequenced. The only difference found in this region between a His- and a weak His+ strain was a single point mutation, an A→G transition. The position of Ty remained unaltered. Examination of lacZ fusion plasmids further implicated this A→G transition as being reponsible for the altered phenotype, the bp transition representing an allele of a cis-acting regulatory element. Subsequent gel retardation and methylation interference experiments revealed that this A→G mutation enabled the binding of a trans-acting factor (TyBf) in vitro. In this paper we show that the TyBf binding site is in a region of chromatin hypersensitive to digestion by DNase I. The binding site is protected in vivo from digestion with exonuclease III, suggesting the presence of a bound protein in His+ (“on”) but not His- (“off”) Ty-containing strains. We propose that a trans-acting factor binding in vivo, presumably TyBf, is responsible for the activation of HIS4 expression in these insertion mutants.
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  • 33
    ISSN: 1432-0983
    Keywords: Yeast ; Mitochondria ; Cytochrome c oxidase ; Post-translational regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Biogenesis of functional cytochrome c oxidase in yeast requires the product of the nuclear gene SCO1. Strains deleted for this gene fail to accumulate the mitochondrially-synthesized cytochrome c oxidase subunits I and II, despite the presence of the respective mRNAs. Here we present data which demonstrate that the observed phenotype does not result from a failure to translate the mRNAs, but from a preferential degradation of the newly synthesized subunits. The SCO1 protein is therefore involved in a post-translational step in the accumulation of cytochrome c oxidase subunits I and II. We propose that the SCO1 protein is required for the correct assembly of both subunits into the cytochrome c oxidase complex.
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  • 34
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    Current genetics 18 (1990), S. 23-27 
    ISSN: 1432-0983
    Keywords: Protein translocation ; Saccharomyces cerevisiae ; Peroxisomes ; Overexpression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Import of proteins into organelles usually requires a cis-acting targeting signal. Analysis of various hybrid proteins, consisting of mouse DHFR and parts of catalase A from Saccharomyces cerevisiae, revealed that fusion proteins containing the N-terminal 126 amino acids, or less, of catalase A remain in the cytosol whereas fusion proteins containing 140, or more, N-terminal amino acids of catalase A form large aggregates inside the cell. These protein bodies, which lack a surrounding membrane, copurified with peroxisomes on cell fractionation. The peroxisomal targeting signal of catalase A does not reside at the C-terminus or at the N-terminus.
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  • 35
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    Mycopathologia 111 (1990), S. 181-189 
    ISSN: 1573-0832
    Keywords: mycotoxin ; ochratoxin ; Penicillium ; storage ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Eleven-kilogram parcels of HY-320 wheat, a cultivar of the new Canada Prairie Spring class, were kept at 15 and 19% initial moisture contents (IMC) in simulated storage in a Manitoba farm granary for 60 weeks to determine biotic and abiotic changes and mycotoxin production. Ochratoxin A reached a maximum of 0.24 ppm by week 20 in the 19% IMC wheat, but was absent in the 15% IMC wheat; no other mycotoxins were detected. Temperature, moisture content, O2 and CO2 levels, fat acidity values, seed germination, microfloral incidence and abundance, and the presence of other mycotoxins were monitored. Principal component analysis of all variables showed that the first principal components accounted for 32–41% of the system variability, and contained the ochratoxin A variable. Ochratoxin A was produced in moist grain that had decreased seed germination andAltermaria activity, and high fungal activity byPenicillium andAspergillus versicolor. Compared to other stored cereals previously studied, HY-320 wheat would be ranked in a low-risk category for mycotoxin formation, based on the ochratoxin A levels observed.
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  • 36
    ISSN: 1573-0832
    Keywords: Nystatin ; amphotericin B ; amphotericin B methyl ester ; polyene antibiotics ; yeast ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Saccharomyces cerevisiae was cultured under anaerobiosis in semi-complete medium to which either palmitoleic or oleic acid was added. Cells were grown at 20 °C or 30 °C. The levels of total lipids, total sterols, and phospholipids were higher in cells grown at 20 °C than at 30 °C. The effects of nystatin (NYS), amphotericin B (AMB), and amphotericin B methyl ester (AME) were evaluated by determining cell viability and liberation of intracellular compounds. The loss of cell viability is higher in the first 30 minutes of incubation with the drugs and is the same regardless of the type of cells obtained. Low molecular weight compounds and ions such as K+ are liberated a few minutes after incubation with the drugs whereas proteins and substances absorbing at 260 nm are liberated later. Phosphate liberation comes after K+ and before compounds of higher molecular weights.
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  • 37
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    Nutrient cycling in agroecosystems 21 (1990), S. 163-166 
    ISSN: 1573-0867
    Keywords: Critical level of Zn ; alkaline soils ; Zn-deficiency ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Field experiments were conducted at 32 locations, chosen for their wide range in DTPA extractable Zn, to determine the critical deficiency level of Zn for predicting response of wheat to Zn application. Soil application of 5.6 kg Zn ha−1 significantly increased the grain yield in deficient soils. Soil extractable Zn was significantly related with per cent grain response and absolute grain yield. Both the graphical and statistical methods of Cate and Nelson indicated the critical level to be 0.75 mg kg−1 soil of DTPA extractable Zn. This level gave a predictability value of 82 per cent.
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  • 38
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    Nutrient cycling in agroecosystems 22 (1990), S. 147-159 
    ISSN: 1573-0867
    Keywords: Sulfate ; elemental sulfur ; leaching ; ryegrass ; wheat ; greenhouse experiments ; split application
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Three factorial experiments with four replications were conducted in a greenhouse to examine the effectiveness of gypsum, elemental sulfur (ES powder) and three S containing N fertilizers, viz., ammonium sulfate (AS), urea + ES, and Ureas (20% AS and 80% urea). All experiments were conducted twice in different years. In the first experiment with uncropped soil, the effects of soil type, leaching rate (2.3 and 6.9 mm water per day) and urea addition on sulfate leaching losses were studied. Leaching losses decreased in the order Ureas 〉 ammonium sulfate (AS) 〉 gypsum ≫ urea + ES. Increasing the leaching rate greatly increased sulfate losses from both soils. Losses were greater in the sandy Typic Hapludoll than in the clayey Oxic Paleustalf. Sulfate adsorption was found to decrease strongly with rising the pH in both soils. Hydrolysis of urea temporarily raised the pH of the soil, thereby increasing the sulfate leaching losses. In the second experiment the effects of S rate (0–65 mg per kg soil), split application and leaching rate (0 and 2.3 mm per day) on sulfate leaching losses and ‘apparent S recovery’ (ASR) by three successive cuts of ryegrass (Lolium perenne L.) were studied. Herbage yield more than doubled when S was applied. The effectiveness of the sulfate fertilizers was greater when S was split-applied than given all at once. With split applications the ASR decreased in the order: Ureas 〉 AS 〉 gypsum 〉 urea + ES 〉 ES powder. ES fertilizers were least effective, because the oxidation rate of ES to sulfate was clearly too slow. In the third experiment the effects of S rate (0–40 mg per kg soil) and split application on sulfate leaching losses and ASR in the grain of wheat (Triticum aestivum L.) were studied under leaching conditions (2.3 mm per day). Grain yield increased strongly due to S application. Split application greatly increased the effectiveness of the sulfate fertilizers and appeared to be an effective tool in satisfying the S need of the crop under leaching conditions. Again, ES fertilizers were least effective, because the oxidation rate of ES was too slow to meet the S demand of the crop. In all experiments leaching losses of sulfate from the ES fertilizers were smaller than from the sulfate fertilizers.
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  • 39
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    Nutrient cycling in agroecosystems 22 (1990), S. 97-107 
    ISSN: 1573-0867
    Keywords: Superphosphate ; placement depth ; banding ; relative effectivenesss ; lupins ; wheat ; field experiment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In a field experiment on a sandplain soil in a low rainfall (326mm per annum) Mediterranean environment of south-western Australia, seven levels of single superphosphate, 0, 7.5, 10, 14, 19.5, 30 and 39 kg P ha−1, were placed at either 3, 5, 7, 9, 11 or 13 cm depth before sowing wheat (Triticum aestivum) at 3 cm. In a separate treatment, superphosphate was drilled with the seed (the normal practice). In the second year, the plots were sown with lupins (Lupinus angustifolius) at 3 cm depth with no additional superphosphate. In three separate treatments, superphosphate at 0, 14 and 39 kg P ha−1, was drilled with the lupin seed (the normal practice) on plots that had received no superphosphate in the first year. Yields of wheat and lupins were used as a measure of the effectiveness of the superphosphate placement treatments relative to the effectiveness of superphosphate drilled with seed of wheat (year 1) or lupins (year 2), to give relative effectiveness (RE) values in each of the two years. In the first year the RE of superphosphate was increased by about 20% when the fertilizer was placed 5 to 9 cm deep in the soil. In the second year, the RE of superphosphate for producing lupin grain was increased by about 30–60% where the fertilizer had been placed 5–13 cm deep in the previous year compared with freshly drilled 3 cm deep. The yield of wheat or lupins was closely related to the P content of plant tissue; each relationship was independent of the depth or year of superphosphate application.
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  • 40
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    Nutrient cycling in agroecosystems 26 (1990), S. 229-235 
    ISSN: 1573-0867
    Keywords: Grass ; wheat ; nitrogen nutrition ; dilution curve ; mineral content ; mineral removal ; phosphorus ; potassium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The important effect of nitrogen in changing the patterns of mineral content and mineral removal is analysed for grass swards and wheat. Different models are proposed; accumulated dry matter developed throughout a growing period is shown to be an excellent reference for assessing the evolution of the plant mineral content and the mineral removal the growing crop. Applications in diagnosing mineral nutrition status and optimising fertilizer use are proposed and discussed.
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  • 41
    ISSN: 1572-9699
    Keywords: 2-Deoxy-D-glucose transport ; polyphosphate ; Saccharomyces cerevisiae ; sugar phosphorylation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The role of polyphosphate in 2-deoxy-D-glucose transport was studied in yeast cells, pulse-labeled with [32P]orthophosphate, by comparing the concentrations and specific activities of polyphosphate, orthophosphate and 2-dGlc-phosphate. When 2-dGlc transport was measured under aerobic conditions, it appeared that polyphosphate replenished the orthophosphate pool, indicating that polyphosphate has, at least mainly, an indirect role in sugar phosphorylation. Also in cells with a reduced respiratory capacity, due to a treatment with antimycin A, no direct role for polyphosphate in 2-dGlc transport could be detected. Under these conditions, only a very limited breakdown of polyphosphate occurred, probably because of the small decrease in the orthophosphate concentration.
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  • 42
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    Cellular and molecular life sciences 46 (1990), S. 193-200 
    ISSN: 1420-9071
    Keywords: Saccharomyces cerevisiae ; protein toxin ; yeast toxin precursor ; protease processing ; lectin ; (1→6)-β-D-glucan ; receptor ; resistant mutants ; spheroplasts ; ion-permeable channels ; site-directed mutagenesis ; toxin functional domains
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The K1 killer toxin ofSaccharomyces cerevisiae is a secreted, virally-coded protein lethal to sensitive yeasts. Killer yeasts are immune to the toxin they produce. This killer system has been extensively examined from genetic and molecular perspectives. Here we review the biology of killer yeasts, and examine the synthesis and action of the protein toxin and the immunity component. We summarise the structure of the toxin precursor gene and its protein products, outline the proteolytic processing of the toxin subunits from the precursor, and their passage through the yeast secretory pathway. We then discuss the mode of action of the toxin, its lectin-like interaction with a cell wall glucan, and its probable role in forming channels in the yeast plasma membrane. In addition we describe models of how a toxin precursor species functions as the immunity component, probably by interfering with channel formation. We conclude with a review of the functional domains of the toxin structural gene as determined by site-directed mutagenesis. This work has identified regions associated with glucan binding, toxin activity, and immunity.
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  • 43
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    Cellular and molecular life sciences 46 (1990), S. 1016-1017 
    ISSN: 1420-9071
    Keywords: In vitro absorption ; calcium ; wheat ; Bengal gram
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The in vitro absorption of calcium from the duodenum was significantly less in a group of rats fed on a wheat diet than in a group fed a wheat and Bengal gram (70∶30) diet.
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  • 44
    ISSN: 1432-072X
    Keywords: Candida boidinii ; Yeast ; Peroxisomes ; β-Oxidation ; d-Amino acid oxidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have studied the induction of peroxisomes in the methylotrophic yeast Candida boidinii by d-alanine and oleic acid. The organism was able to utilize each of these compounds as the sole carbon source and grew with growth rates of μ=0.20 h-1 (on d-alanine) or μ=0.43 h-1 (on oleic acid). Growth was associated with the development of many peroxisomes in the cells. On d-alanine a cluster of tightly interwoven organelles was observed which made up 6.3% of the cytoplasmic volume and were characterized by the presence of d-amino acid oxidase and catalase. On oleic acid rounded to elongated peroxisomes were dominant which were scattered throughout the cytoplasm. These organelles contained increased levels of β-oxidation enzymes; their relative volume fraction amounted 12.8% of the cytoplasmic volume.
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  • 45
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    Archives of microbiology 154 (1990), S. 267-273 
    ISSN: 1432-072X
    Keywords: Yeast ; Saccharomyces cerevisiae ; (R)-2,3-Butanediol dehydrogenase ; Stereospecificity ; Gas chromatographic analysis of enantiomers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A NAD-dependent (R)-2,3-butanediol dehydrogenase (EC 1.1.1.4), selectively catalyzing the oxidation at the (R)-center of 2,3-butanediol irrespective of the absolute configuration of the other carbinol center, was isolated from cell extracts of the yeast Saccharomyces cerevisiae. Purification was achieved by means of streptomycin sulfate treatment, Sephadex G-25 filtration, DEAE-Sepharose CL-6B chromatography, affinity chromatography on Matrex Gel Blue A and Superose 6 prep grade chromatography leading to a 70-fold enrichment of the specific activity with 44% yield. Analysis of chiral products was carried out by gas chromatographic methods via pre-chromatographic derivatization and resolution of corresponding diasteromeric derivatives. The enzyme was capable to reduce irreversibly diacetyl (2,3-butanediol) to (R)-acetoin (3-hydroxy-2-butanone) and in a subsequent reaction reversibly to (R,R)-2,3-butanediol using NADH as coenzyme. 1-Hydroxy-2-ketones and C5-acyloins were also accepted as substrates, whereas the enzyme was inactive towards the reduction of acetone and dihydroxyacetone. The relative molecular mass (M r) of the enzyme was estimated as 140 000 by means of gel filtration. On SDS-polyacrylamide gel the protein decomposed into 4 (identical) subunits of M r 35 000. Optimum pH was 6.7 for the reduction of acetoin to 2,3-butanediol and 7.2 for the reverse reaction.
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  • 46
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    Archives of microbiology 154 (1990), S. 175-178 
    ISSN: 1432-072X
    Keywords: Killer toxin ; Hanseniaspora uvarum ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The yeast Hanseniaspora uvarum liberates a killer toxin lethal to sensitive strains of the species Saccharomyces cerevisiae. Secretion of this killer toxin was inhibited by tunicamycin, an inhibitor of N-glycosylation, although the mature killer protein did not show any detectable carbohydrate structures. Culture supernatants of the killer strain were concentrated by ultrafiltration and the extracellular killer toxin was precipitated with ethanol and purified by ion exchange chromatography. SDS-PAGE of the electrophoretically homogenous killer protein indicated an apparent molecular mass of 18,000. Additional investigations of the primary toxin binding sites within the cell wall of sensitive yeast strains showed that the killer toxin of Hanseniaspora uvarum is bound by β-1, 6-d-glucans.
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  • 47
    ISSN: 1432-072X
    Keywords: 13C NMR spectroscopy ; Yeast ; Debaryomyces hansenii ; Osmoregulation ; Compatible solute ; Salt stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract High resolution 13C NMR combined with chemical analysis were used to study the formation of metabolites from [1-13C]-labelled glucose by the salt-tolerant yeast Debaryomyces hansenii after transfer to media containing 8% NaCl. Time course spectroscopy of an aerobic cell suspension showed [1,3-13C]glycerol as the predominant end product. Perchloric acid extracts revealed additional less prominent incorporation of label into arabinitol, trehalose, glutamic acid, and alanine. The incorporation into trehalose and arabinitol showed a transient increase after shift to the high salinity medium. It is concluded that glycerol and arabinitol are the major organic solutes in D. hansenii, the production of glycerol being strongly induced by high salinity. Analysis of labelled extracts of D. hansenii after transfer to 8% NaCl media containing [1-13C]- or [6-13C]glucose, demonstrated that glucose is dissimilated via a combination of the Embden-Meyerhof-Parnas pathway and the pentose phosphate pathway, with the former playing a major role in glycerol formation and the latter in arabinitol production. The almost exclusive labelling of C5 of arabinitol from [6-13C]glucose indicates that the pathway to arabinitol proceeds via reduction of ribulose-5-phosphate.
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  • 48
    ISSN: 1432-072X
    Keywords: cAMP ; Cat mutants ; Glucose repression ; Glucose-induced ; Intracellular pH ; Ras ; Saccharomyces cerevisiae ; Signal transduction ; Trehalase ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Addition of glucose to derepressed cells of the yeast Saccharomyces cerevisiae induces a transient, specific cAMP signal. Intracellular acidification in these cells, as caused by addition of protonophores like 2,4-dinitrophenol (DNP) causes a large, lasting increase in the cAMP level. The effect of glucose and DNP was investigated in glucose-repressed wild type cells and in cells of two mutants which are deficient in derepression of glucose-repressible proteins, cat1 and cat3. Addition of glucose to cells of the cat3 mutant caused a transient increase in the cAMP level whereas cells of the cat1 mutant and in most cases also repressed wild type cells did not respond to glucose addition with a cAMP increase. The glucose-induced cAMP increase in cat3 cells and the cAMP increase occasionally present in repressed wild type cells however could be prevented completely by addition of a very low level of glucose in advance. In derepressed wild type cells this does not prevent the specific glucose-induced cAMP signal at all. These results indicate that repressed cells do not show a true glucose-induced cAMP signal. When DNP was added to glucose-repressed wild type cells or to cells of the cat1 and cat3 mutants no cAMP increase was observed. Addition of a very low level of glucose before the DNP restored the cAMP increase which points to lack of ATP as the cause for the absence of the DNP effect. These data show that intracellular acidification is able to enhance the cAMP level in repressed cells. The glucose-induced artefactual increase occasionally observed in repressed cells is probably caused by the fact that their low intracellular pH is only restored after the ATP level has increased to such an extent that it is no longer limiting for cAMP synthesis. It is unclear why the artefactual increases are not always observed. Measurement of glucose- and DNP-induced activation of trehalase confirmed the physiological validity of the changes observed in the cAMP level. Our results are consistent with the idea that the glucose-induced signaling pathway contains a glucose-repressible protein and that the protein is located before the point where intracellular acidification triggers activation of the pathway.
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  • 49
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Catalase A ; Catalase T ; β-Oxidation ; Microbodies ; H2O2-Metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The parental strain (A+T+) of Saccharomyces cerevisiae and mutants, deficient in catalase T (A+T−), catalase A (A−T+) or both catalases (A−T−), grew on ethanol and oleic acid with comparable doubling times. Specific activities of catalase were low in glucose- and ethanol-grown cells. In the two oleic acid-grown A+-strains (A+T+ and A+T−) high catalase activities were found; catalase activity invariably remained low in the A−T+ strain and was never detected in the A−T− strain. The levels of β-oxidation enzymes in oleic acid-grown cells of the parental and all mutant strains were not significantly different. However, cytochrome C peroxidase activity had increased 8-fold in oleic acid grown A− strains (A−T+ and A−T−) compared to parental strain cells. The degree of peroxisomal proliferation was comparable among the different strains. Catalase A was shown to be located in peroxisomes. Catalase T is most probably cytosolic in nature and/or present in the periplasmic space.
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  • 50
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    Archives of microbiology 153 (1990), S. 384-391 
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Ethanol ; Acetic acid ; Cytoplasmic pH ; 31P-NMR ; 13C-NMR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cell suspensions of a respiratory deficient mutant of Saccharomyces cerevisiae were monitored by in vivo 31P and 13C Nuclear Magnetic Resonance in order to evaluate the effect of ethanol in intracellular pH and metabolism. In the absence of an added energy source, ethanol caused acidification of the cytoplasm, as indicated by the shift to higher field of the resonance assigned to the cytoplasmic orthophosphate. Under the experimental conditions used this acidification was not a consequence of an increase in the passive influx of H+. With cells energized with glucose, a lower value for the cytoplasmic pH was also observed, when ethanol was added. Furthermore, lower levels of phosphomonoesters were detected in the presence of ethanol, indicating that an early event in glycolysis is an important target of the ethanol action. Acetic acid was identified as responsible for the acidification of the cytoplasm, in experiments where [13C]ethanol was added and formation of labeled acetic acid was detected. The intracellular and the extracellular concentrations of acetic acid were respectively, 30 mM and 2 mM when 0.5% (120 mM) [13C]ethanol was added.
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  • 51
    ISSN: 1432-072X
    Keywords: Regulatory mutants ; Meiotic mapping ; Transcriptional regulation ; MAL genes ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The MAL1 locus of Saccharomyces cerevisiae comprises three genes necessary for maltose utilization: a regulatory (MALR), a maltose transport (MALT) and a maltase gene (MALS). A fine structure genetic map of the MAL1R gene was constructed and the order of mutations was confirmed by plasmid-mediated chromosomal recombination. The mutations cluster non-randomly within the 5′ half of the gene, where the putative DNA binding domain of the encoded protein is located. Only mutations mal1 R-22 and MAL1R-72 map in the 3′ terminal half of the gene; these mutations cause a different pattern of transcriptional regulation of plasmid-borne MAL6T genes. Experiments supporting a direct involvement of the MALR-encoded protein in carbon catabolite repression of MAL gene expression are reported.
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  • 52
    ISSN: 1573-4919
    Keywords: bovine heart fatty acid-binding protein ; H-FABPc ; heterologous gene expression ; Saccharomyces cerevisiae ; GALIO promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Summary The unicellular eukaryotic microorganism, Saccharomyces cerevisiae, transformed with a plasmid containing a cDNA fragment encoding bovine heart fatty acid-binding protein (H-FABP) under the control of the inducible yeast GAL10 promoter, expressed FABP during growth on galactose. The maximum level of immunoreactive FABP, identical in size to native protein as judged from SDS-polyacrylamide gel electrophoresis, was reached after approximately 16 hours of induction. Analysis of particulate and soluble subcellular fractions showed that FABP was exclusively associated with the cytosol. FABP expressed in yeast cells was functional as was demonstrated by its capacity to bind 14C-oleic acid in an in vitro assay. Growth of the transformants on galactose as the carbon source was significantly retarded at 37°C. Whereas the fatty acid pattern of total lipids was not altered in transformed cells, desaturation of exogenously added 14C-palmitic acid was significantly reduced both at 30 and 37°C. The lowest percentage of radioactively labeled unsaturated fatty acids was found in the phospholipid fraction.
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  • 53
    ISSN: 1573-1561
    Keywords: Allelochemicals ; no-tillage ; conventional-tillage ; soils ; wheat ; Triticum aestivum ; mass spectrometry ; Petri-dish bioassay ; fatty acids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Putative allelochemicals found in the soil of no-tillage and conventional-tillage wheat plots near Stillwater, Oklahoma, were obtained by a mild alkaline aqueous extraction procedure, bioassayed to determine their biological activity, purified, and analyzed with a capillary gas chromatography-mass spectrometry-data analysis system. The most significant inhibition was found in bioassays of extracts from soil collected immediately after harvest in June, July, and August. No-tillage soils produced significant inhibition during the rest of the year also. Mass spectrometry showed fatty acids as the most abundant compounds. However, when bioassayed authentic samples of the five free fatty acids showed no significant biological activity toward wheat.
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  • 54
    ISSN: 1573-5028
    Keywords: tRNA-like sequences ; t-elements ; RNA processing ; mitochondria ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have recently described the properties of a wheat mitochondrial extract that is able to process, accurately and efficiently, artificial transcripts containing wheat mitochondrial tRNA sequences, with the production of mature tRNAs (P.J. Hanic-Joyce and M.W. Gray, J. Biol. Chem., in press). Such processing involves 5′-endonucleolytic, 3′-endonucleolytic, and TRNA nucleotidyltransferase activities. Here we show that this system also acts on transcripts containing sequences corresponding to an unusual class of short repeats (‘t-elements’) in wheat mtDNA. These repeats are theoretically capable of assuming a tRNA-like secondary structure, although stable transcripts corresponding to them are not detectable in vivo. We find that t-element sequences are processed with the same specificity and with comparable efficiency as are authentic tRNA sequences. Because known t-elements are located close to and in the same transcriptional orientation as active genes (18S-5S, 26S, tRNAPro) in wheat mtDNA, our results raise the question of whether t-elements play a role in gene expression in wheat mitochondria.
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  • 55
    ISSN: 1573-5028
    Keywords: α-amylase tetrameric inhibitor ; cDNA cloning ; genetic mapping ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have characterized three cDNA clones corresponding to proteins CM1, CM3 and CM16, which represent the three types of subunits of the wheat tetrameric inhibitor of insect α-amylases. The deduced amino acid sequences of the mature polypeptides are homologous to those of the dimeric and monomeric α-amylase inhibitors and of the trypsin inhibitors. The mature polypeptides are preceded by typical signal peptides. Southern blot analysis of appropriate aneuploids, using the cloned cDNAs as probes, has revealed the location of genes for subunits of the CM3 and of the CM16 type within a few kb of each other in chromosomes 4A, 4B and 4D, and those for the CM1 type of subunit in chromosomes 7A, 7B and 7D. Known subunits of the tetrameric inhibitor corresponding to genes from the B and D genomes have been previously characterized. No proteins of this class have been found to be encoded by the A genome in hexaploid wheat (genomes AA, BB, DD) or in diploid wheats (AA) and no anti α-amylase activity has been detected in the latter, so that the A-genome genes must be either silent (pseudogenes) or expressed at a much lower level.
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  • 56
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    Plant molecular biology 15 (1990), S. 793-795 
    ISSN: 1573-5028
    Keywords: mitochondrial gene ; ORF25 ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The wheat mitochondrial orf25 nucleotide sequence of 576 pb has been determined. Its derived protein sequence shares 88% and 75% amino acid identity with those of maize and tobacco mitochondria, respectively. The wheat and tobacco orf25 sequences lack four inserts, of 6 bp to 36 bp, that are present in the maize homologue. The wheat orf25 gene is actively transcribed and is preceded by a regulatory sequence block very similar to those located upstream of the wheat coxII and atp6 genes. Our observations support the view that orf25 sequences encode a functional polypeptide in plant mitochondria.
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  • 57
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    The journal of membrane biology 116 (1990), S. 93-105 
    ISSN: 1432-1424
    Keywords: clathrin ; genetics ; Saccharomyces cerevisiae ; exocytosis ; endocytosis ; prohormone maturation
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    Topics: Biology , Chemistry and Pharmacology
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  • 58
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    Theoretical and applied genetics 79 (1990), S. 305-313 
    ISSN: 1432-2242
    Keywords: Peroxidase ; Isoelectric focusing ; Hexaploid ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Isoelectric focusing (IEF) of extracts from different tissues of hexaploid wheat cv “Chinese Spring” provided a method of distinguishing and identifying the four known, and one newly discovered, sets of genes encoding peroxidase isozyme production.Per-1, carried on the short arms of homoeologous group 1 chromosomes, shows a high degree of conservation and is active in coleoptile tissue.Per-2, carried on the short arms of group 2 chromosomes, shows some polymorphism and is most active in root tissue.Per-3, on the long arms of group 3 chromosomes, is highly variable and most active in embryo tissue.Per-4, carried on chromosome arms7AS,4AL, and7DS, is quite variable and most active in endosperm tissue. (The chromosome nomenclature used in this paper is that agreed to by the 7th International Wheat Genetics Symposium, where the previous designations of4A and4B were reversed.) Restriction fragment length polymorphism (RFLP)-based maps of the group 7 chromosomes were used to locatePer-A4 to a distal region of7AS. In addition, a further set of genes was identified as being active in root tissue. In wheat a single locus,Per-D5, was found on chromosome arm2DS.
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  • 59
    ISSN: 1573-5036
    Keywords: Lycopersicon esculentum L. ; magnesium-manganese interaction ; magnesium: manganese ratio ; manganese toxicity ; tomato ; Triticum aestivum L. ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Results are reported for tomato (Lycopersicon esculentum L. var. Ailsa craig) and wheat (Triticum aestivum L. cv. Mara) which demonstrate that increasing concentrations of Mg in the plant raises plant tolerance to Mn toxicity. Water culture experiments with tomato show that under conditions of high Mn supply (200 µM, Mn), not only does increasing Mg application (0.75 mM to 15 mM) depress Mn uptake, but the higher Mg concentrations in the shoot counteract the onset of Mn toxicity when the concentrations of Mn in the shoot are also high. The ratio of Mg: Mn in the tissues is a better indicator of the appearance of toxicity symptoms than Mn concentration alone. Toxicity symptoms were observed when the Mg:Mn ratio in the shoot tissue was from 1.13 to a value between 3.53 and 6.54. The corresponding Mg: Mn ratio in the older leaves was from 0.82 to between 2.27 and 3.51. For wheat grown in soil, analyses of leaves revealed that growth could be expressed by the following relationship: Y=A+B exp(-kX), where Y=growth, X=Mg:Mn ratio, A, B and k=constants. Growth was significantly reduced when the Mg:Mn ratio fell below 20:1. From a measurement of this ratio it is therefore possible to predict the appearance of Mn toxicity and its influence on growth.
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  • 60
    ISSN: 1573-5036
    Keywords: calcium ; ear development ; magnesium ; potassium ; tissue press sap ; Triticum aestivum L. ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract For floret development and final grain number ear elongation in wheat (10–17d before anthesis) is the most sensitive period to adverse growth conditions. Sugars are supposedly the main factor determining floret numbers and grain set, and play next to potassium a significant role in establishing turgor pressure in young tissues. In view of this osmotic function, the influence of K on the concentration of water soluble carbohydrates (WSC, total of sucrose and reducing sugars), Ca and Mg was investigated in pot experiments. Further, the osmotic potential of sap extracted from young ears was determined and compared to sap extracted from the leaf blade. Plants supplied with low amounts of K (moderate K-deficiency) had a considerably lower K concentration in the press sap of the flag leaf and the ear than plants well supplied with K. Concentrations of WSC, Ca and Mg were higher in press sap of the flag leaf in K deficient plants than in plants adequately supplied with K. This indicates a substitution of K in its osmotic role. In press sap from ears, however, WSC, Ca and Mg were not influenced by the K application. Therefore, substances other than those measured must have been responsible for the osmoregulation in the young ear. WSC and the osmotic potential increased (more negative) independently on K supply during ear elongation, while K, Ca and Mg concentrations decreased. Whereas grain number was not influenced by the treatment, single grain weight at maturity was reduced by low K availability in the soil.
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  • 61
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    Plant and soil 124 (1990), S. 303-307 
    ISSN: 1573-5036
    Keywords: models ; potassium uptake ; Triticum aestivum L. ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Spring wheat was grown in the field under deficient and sufficient levels of soil K and with high and low supplies of fertiliser nitrogen. Measurements were made of K uptake, soil nutrient supply parameters, root growth and, in solution culture, root influx parameters. Mechanistic models predicted uptake reasonably well under K-deficient conditions, but over-predicted uptake, by as much as 4 times, under K-sufficient conditions. The over-prediction was apparently due to poor characterisation of plant demand.
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  • 62
    ISSN: 1573-5036
    Keywords: Cu levels ; N sources ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A pot experiment was conducted, in a greenhouse, at Hisar, India, using a sandy soil deficient in nitrogen and copper, to study the effects of various levels of N and Cu on the dry matter yield and the N and Cu contents of wheat (Triticum aestivum L.). The sources of nitrogen used were Ca(NO3)2, NH4Cl and NH4NO3 applied in amounts necessary to establish 120 ppm of soil nitrogen and using a control (0 ppm N). Copper was applied, as copper chloride, to give soil Cu levels of 0, 5, 10 and 20 ppm. In general, dry matter yields, N and Cu concentrations in shoots and roots and available soil-N after harvest of the plants, followed the order Ca(NO3)〉NH4NO3〉NH4Cl. Up to a level of 5 ppm Cu, the dry matter yields of shoots and roots increased, but decreased at higher levels of Cu. Increasing Cu levels significantly decreased the available soil-nitrogen after harvest and also the concentration of N in the plants. At the same time the concentration of Cu in shoots and roots and available Cu in the soil was increased. Nitrogen and copper were found to have a mutually antagonistic effect on each other's concentration in the plants. The antagonism was greater with NH4 + sources than with NO3 − compounds.
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  • 63
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    European journal of plant pathology 96 (1990), S. 187-198 
    ISSN: 1573-8469
    Keywords: fusarium head blight ; Fusarium culmorum ; F. graminearum ; wheat ; mycotoxin ; deoxynivalenol ; nivalenol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Description / Table of Contents: Samenvatting Aaraantasting van tarwe doorFusarium culmorum enFusarium graminearum leidt tot vorming van mycotoxinen in het graan, waarvan deoxynivalenol (DON) en nivalenol (NIV) de belangrijkste toxinen zijn. In dit artikel wordt een overzicht gegeven van de toxicologische aspecten, en het voorkomen van deze toxinen in tarwe. Informatie over DON en NIV in tarwe in West-Europa is schaars. Gebaseerd op gegevens vanFusarium epidemieën in de jaren 1979–1986 wordt een schatting gegeven van de concentratie DON in Nederlandse tarwe. Rekening houdend met de herkomst en verwerking van tarwe, blijken zowel in dierlijk als menselijk voedsel lage concentraties DON chronisch voor te komen. Op basis van een maximaal toelaatbare dagelijkse dosis DON van 3 μg kg−1 lichaamsgewicht is de schatting van de dagelijkse opname van DON in het jaar volgend op de oogst van 1982 net op de grens. Zowel een jaarlijkse inventarisatie vanFusarium aantasting en DON besmetting van het graan, als de ontwikkeling vanFusarium-resistente rassen zijn noodzakelijk.
    Notes: Summary An infection of bread wheat by fusarium head blight contaminates the crop with mycotoxins, particularly deoxynivalenol (DON) and nivalenol (NIV). The toxicity and natural occurrence of these mycotoxins in wheat are reviewed. Based on 8 years data of fusarium head blight epidemics of wheat in the Netherlands, DON contamination of the grain was estimated. Fusarium head blight ratings averaged an infection of 1.7% of all spikelets; estimates for DON contamination averaged 0.9 mg kg−1. Taking a guideline level for DON in uncleaned bread wheat of 2 mg kg−1, in 1979 and 1982 a wheat crop was produced with estimated DON concentrations above the limit of tolerance. Human and animal exposure to mycotoxins in the Netherlands appears to be small but chronic. The information presented in this paper illustrates the need for an annual evaluation of the crop for fusarium head blight incidence and mycotoxin content, and the necessity of fusarium head blight resistant wheat cultivars.
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  • 64
    ISSN: 1617-4623
    Keywords: Ty elements ; Saccharomyces cerevisiae ; Retrotransposon
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary To learn more about the variety of Ty elements capable of activating gene expression, we characterized 206 spontaneous Ty transpositions that activate the promoterless gene his3Δ4. Most of the Ty elements appear to be full-length, although a few deleted elements were recovered. Over 95% of the insertions belong to the Ty1 family, and the rest are Ty2 elements. The excessive number of Ty1 transpositions was unexpected because there are only 2-fold more Ty1 than Ty2 elements in the yeast strains used in the selection. However, there is 20-fold more Ty1 than Ty2 RNA present in these yeast strains. This difference in RNA level explains the greater number of Ty1 verses Ty2 transpositions at his3Δ4, because Ty elements transpose through an RNA intermediate. A similar association between the Ty transcript level and transpositional activation of his3Δ4 is obtained in cells expressing GAL1-promoted Ty2-H556 or Ty2-917 elements, but only if the element does not contain a marker. Genetically marked Ty2-H556NEO and-917NEO elements transpose into and activate his3Δ4 with the same efficiency as the previously characterized Ty1H3NEO element, but are underrepresented relative to the levels of TyNEO transcript. We also found that chromosomal Ty transcripts are even more abundant than previously estimated and comprise about 1% of total cellular RNA.
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  • 65
    ISSN: 1617-4623
    Keywords: Yeast ; Cell cycle ; Meiosis ; DNA primase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mitotic cultures synchronised either by a feed-starve protocol or by elutriation have been used to show that the Saccharomyces cerevisiae DNA primase I gene is periodically expressed in the cell cycle. The transcript increases many-fold in late G1 and reaches a peak at the same time as four other genes essential for DNA synthesis, CDC8, CDC9, CDC21 and POL1. The primase I transcript is also regulated in meiosis, reaching maximal levels during premeiotic DNA synthesis.
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  • 66
    ISSN: 1617-4623
    Keywords: Bacterial conjugation ; Cyanobacteria ; IncQ plasmids ; Saccharomyces cerevisiae ; DNA methylation
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    Topics: Biology
    Notes: Summary The promiscuous IncQ plasmid pKT210 (Cmr, Smr) is efficiently transferred by transpecific conjugation from Escherichia coli to the facultatively heterotrophic cyanobacterium Synechocystis PCC6803 when mobilized by a helper plasmid coding for IncP transfer functions. The IncQ plasmid is stably maintained in the cyanobacterium as an autonomously replicating multicopy plasmid with no detectable structural alterations and can be recovered by transformation back to E. coli when using a mcrA mcrB host. Thus, the replicative host-range of IncQ plasmids extends beyond purple bacteria to the distinct procaryotic taxon of cyanobacteria, allowing the use of these small plasmids as convenient cloning vectors in Synechocystis PCC6803 and presumably also in cyanobacteria that are not amenable to genetic transformation. In contrast, an IncQ plasmid bearing the TRP1 gene of Saccharomyces cerevisiae failed to replicate when transferred to that yeast by transformation.
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  • 67
    ISSN: 1617-4623
    Keywords: Meiosis ; Sporulation ; Northern hybridization ; Regulatory circuit ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary SME1 was cloned due to its high copy number effect: it enabled MATα/MATα diploid cells to undergo meiosis and sporulation in a vegetative medium. Disruption of SME1 resulted in a recessive Spo− phenotype. These results suggest that SME1 is a positive regulator for meiosis. DNA sequencing analysis revealed an open reading frame of 645 amino acids. An amino terminal peptide of ca 400 amino acids in the deduced protein was similar to known protein kinases. Transcription of SME1 was regulated negatively by nitrogen and glucose and positively by MATα/MATα and IME1, another positive regulator gene of meiosis. By complementation analysis, SME1 was found to be identical to IME2, which had been shown to be important in meiosis. These results suggest that IME1 product stimulates meiosis by activating transcription of SME1 (IME2) and that protein phosphorylation is required for initiation of meiosis.
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  • 68
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; AR07 gene ; Chorismate mutase ; GCN4 ; Transcriptional regulation
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary The gene AR07 encodes the monofunctional enzyme chorismate mutase, a branch point enzyme in the aromatic amino acid biosynthetic pathway in Saccharomyces cerevisiae. We investigated the transcription of the AR07 gene. Three 5′ ends at positions − 36, − 56 and − 73 and the 3′ end of the transcripts 146 bp downstream of the translational stop codon were mapped. As in the promoters of other aromatic amino acid biosynthetic genes, a recognition element for the GCN4 transcriptional activator of amino acid biosynthesis is located 425 base pairs (bp) upstream of the first transcriptional start point. This element binds GCN4 specifically in vitro. Northern analysis and determination of the specific enzyme activity reveals however, that the element is not sufficient to mediate transcriptional regulation by GCN4 in vivo. We thus suggest that in addition to a consensus sequence capable of binding the GCN4 protein other factors like, for example, chromatin structure, determine whether a recognition site for a transcription factor functions as an upstream activation sequence.
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  • 69
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Splicing ; Nuclear structure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We report on the characterization of the yeast prp20-1 mutant. In this temperature-sensitive mutant, multiple steps of mRNA metabolism are affected. The prp20-1 mutant strain showed alterations in mRNA steady-state levels, defective mRNA splicing and changes in transcription initiation or termination when shifted from the permissive to the non-permissive temperature. In addition, a change in the structure of the nucleus in these cells became apparent. Electron microscopy revealed an altered structure of the nucleoplasm of prp20-1 mutant cells when grown at the no-permissive temperature that was not observed in cells grown at the permissive temperature or in wild-type cells. The wild-type PRP20 gene was isolated and sequenced. The putative PRP20 protein has a molecular weight of 52 kDa. We found that the PRP20 gene is identical to the yeast SRM1 gene (Clark and Sprague 1989). In addition, the PRP20 protein sequence shows significant sequence similarity to the human RCC1 protein (Ohtsubo et al. 1987). This protein has been implicated in the control of chromosome condensation. Based on the phenotype of the prp20-1 mutant and the observed sequence similarity to the human RCC1 protein, we postulate that the yeast PRP20 protein is involved in the control of nuclear organization.
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  • 70
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    Molecular genetics and genomics 224 (1990), S. 111-118 
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Translation ; Splicing ; Paromomycin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The MSS51 gene product has been previously shown to be involved in the splicing of the mitochondrial pre-mRNA of cytochrome oxidase subunit I (COX1). We show here that it is specifically required for the translation of the COX1 mRNA. Furthermore, the paromomycin-resistance mutation (P inf454 supR ) which affects the 15 S mitoribosomal RNA, interferes, directly or indirectly, with the action of the MSS51 gene product. Possible roles of the MSS51 protein on the excision of COX1 introns are discussed.
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  • 71
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    Molecular genetics and genomics 224 (1990), S. 209-221 
    ISSN: 1617-4623
    Keywords: Aminoacyl-tRNA synthetase ; RNA splicing ; Group I introns ; RNA maturase ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The Saccharomyces cerevisiae nuclear gene NAM2 codes for mitochondrial leucyl-tRNA synthetase (mLRS). Herbert et al. (1988, EMBO J 7:473–483) proposed that this protein is involved in mitochondrial RNA splicing. Here we present the construction and analyses of nine mutations obtained by creating two-codon insertions within the NAM2 gene. Three of these prevent respiration while maintaining the mitochondrial genome. These three mutants: (1) display in vitro a mLRS activity ranging from 0%–50% that of the wild type: (2) allow in vivo the synthesis of several mitochondrially encoded proteins; (3) prevent the synthesis of the COXII protein but not of its mRNA; (4) abolish the splicing of the group I introns bI4 and aI4; and (5) affect significantly the excision of the group I introns bI2, bI3 and aI3. Importation of the bI4 maturase from the cytoplasm into mitochondria in a nam2 − mutant strain does not restore the excision of the introns bI4 and aI4 implying that the splicing deficiency does not result from the absence of the bI4 maturase. We conclude that the mLRS is a splicing factor essential for the excision of the group I introns bI4 and aI4 and probably important for the excision of other group I introns.
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  • 72
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    Molecular genetics and genomics 224 (1990), S. 257-263 
    ISSN: 1617-4623
    Keywords: General Control ; Thermotolerance ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In Saccharomyces cerevisiae starvation for a single amino acid activates the transcription of a set of genes belonging to different amino acid biosynthetic pathways (General Control, GC). We show that mutants affected in GC regulation are also affected in their response to thermal stress. Moreover, growth conditions that are known to induce heat shock proteins induce the GC response. However, unlike heat shock proteins, the transcriptional activator of GC, GCN4, is not induced after a short exposure to heat, and in gcn mutant strains induction of heat resistance is normal.
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  • 73
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    Molecular genetics and genomics 220 (1990), S. 269-276 
    ISSN: 1617-4623
    Keywords: GABA ; Saccharomyces cerevisiae ; Transcription ; DNA sequence ; Zinc finger
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The UGA3 gene of Saccharomyces cerevisiae is required for 4-aminobutyric acid (GABA)-dependent induction of the UGA1, UGA2 and UGA4 genes which encode the two GABA catabolic enzymes and a GABA-specific permease, respectively. Measurements of UGA1-specific transcripts show that induction of UGA1 correlates with accumulation of its RNA and requires a functional UGA3 gene. A 2 kb DNA fragment complementing the uga3 mutation was isolated and shown to contain the UGA3 gene. The primary structure of the UGA3 encoded protein was deduced from the DNA sequence, and contains an N-terminal, cysteine-rich motif similar in sequence to regions found in other fungal regulatory proteins and which are supposed to form zinc finger structures involved in DNA binding. Mutations were identified in the UGA3 genes isolated from uninducible and constitutive uga3 alleles. One case of intragenic complementation between two uninducible uga3 mutants is reported, indicating a possible oligomeric structure for UGA3. The role of UGA3 is discussed in relation to its genetic properties and its predicted structure.
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  • 74
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    Molecular genetics and genomics 220 (1990), S. 283-288 
    ISSN: 1617-4623
    Keywords: Chorismate mutase ; Saccharomyces cerevisiae ; Repression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The Saccharomyces cerevisiae ARO7 gene was cloned by screening a wild-type gene bank for complementation of an aro7 auxotrophic mutant. In vitro mutagenesis of the isolated plasmid (pJFB1) gave several transformants resistant to levels of the phenylalanine analogue 2-thienylalanine inhibitory to the wild-type transformant. Chorismate mutase assays indicated that two of the mutants (J14-26IV6 and J14-26IV9) were resistant to feedback inhibition by tyrosine displayed by wild-type strains. Analysis of the effect of other aromatic amino acids on chorismate mutase activity showed that tryptophan counteracted this inhibition. Analysis of the effect of tyrosine in the growth medium on enzyme activity indicated that the wild-type ARO7 gene was repressed by tyrosine, a phenomenon not previously reported. Two of the 2-thienylalanine resistant mutants (J14-26IV3 and J14-26IV9) appeared to be resistant to this repression. Transcriptional analysis confirmed that the level of ARO7 transcript decreased with increasing tyrosine concentration. In stain J14-26IV9 the ARO7 transcript level was not affected. J14-26IV9, therefore, appears to be a double mutant, resistant to both feedback inhibition and repression by tyrosine.
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    Molecular genetics and genomics 222 (1990), S. 87-96 
    ISSN: 1617-4623
    Keywords: Glucoamylase ; Sporulation ; Saccharomyces cerevisiae ; Saccharomyces cerevisiae var. diastaticus ; STA genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The single glucoamylase gene (SGA1) of the yeast Saccharomyces cerevisiae is expressed exclusively during the sporulation phase of the life cycle. Enzymatic studies and nucleic acid sequence comparisons have shown that the SGA1 glucoamylase is closely related to the secreted enzymes of S. cerevisiae var. diastaticus. The latter are encoded by any of three unlinked STA genes, which have been proposed to derive from the ancestral SGA1 form by genomic rearrangement. We show that the regulation of SGA1 is distinct from that of the other members of the STA gene family. SGA1 expression did not respond to STA10, the primary determinant of glucoamylase expression from STA2. Unlike STA2, SGA1 was not regulated directly by the mating type locus. Expression of SGA1 depended on the function of the MAT products in supporting sporulation and not on the formation of haploid progeny spores or on the composition of the mating type locus per se. We conclude that the STA genes acquired regulation by STA10 and MAT by the genomic rearrangements that led to their formation. This regulation is thus distinct from that of the ancestral SGA1 gene.
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  • 76
    ISSN: 1617-4623
    Keywords: General amino acid permease ; Protein kinase ; Serine-rich protein ; Transport protein ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary TheNPR1 gene ofSaccharomyces cerevisiae plays a central role in controlling permease activity; its product is required to promote the activity of at least six distinct transport systems for nitrogenous nutrients under conditions of nitrogen catabolite derepression. We report here the nucleotide sequence of the clonedNPR1 gene. The predicted amino acid sequence indicates thatNPR1 encodes a protein of 86 kDa which appears to be organized into two distinct structural domains. The amino-terminal domain of NPR1 (residues 1 to 440) contains 26% serine residues and several regions strongly enriched for PEST residues suggesting a short half-life for the NPR1 protein. The carboxy-terminal region of NPR1 contains consensus sequences characteristic of the catalytic domains of protein kinases. Therefore, NPR1-dependent positive control of nitrogen transport systems most likely involves protein phosphorylation. Northern analysis indicates that the absence of general amino acid permease (GAP1) activity innpr1 mutants is not due to reduction in transcription or messenger stability. Hence, the NPR1 protein probably acts at the post-transcriptional level. Proteins that may serve as substrates for phosphorylation are discussed.
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  • 77
    ISSN: 1617-4623
    Keywords: Yeast ; Arginine ; Regulatory protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We present here a functional analysis of ARGRI and ARGRIII regulatory proteins which are involved together with ARGRII in specific regulation of arginine anabolic and catabolic pathways. Unlike ARGRII, ARGRI and ARGRIII have no transcriptional activation capacity. The first 60 amino acids of ARGRI (out of 177) are dispensable for its activity. The functional domain of the protein is located in the region of homology with MCM1 and SRF proteins. ARGRIII contains in its C-terminal portion a stretch of 17 aspartate residues which are indispensable for arginine regulation. Gene disruption of theARGRIII gene impairs the growth of the mutant on rich medium, showing that ARGRIII has a pleiotropic role in the cell.
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  • 78
    ISSN: 1617-4623
    Keywords: Aspergillus nidulans ; Schizosaccharomyces pombe ; Saccharomyces cerevisiae ; Drug resistance ; β-tubulin mutation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Rhizoxin, an antibiotic, exhibits potent anti-mitotic activity against most eucaryotic cells including those of higher vertebrates, plants and fungi by binding to β-tubulin. ThebenA gene of three independently isolated rhizoxin-resistant (Rhir) mutants ofAspergillus nidulans was cloned, sequenced and compared with that of the wild-type, rhizoxin-sensitive (Rhis) strain. In all three Rhir mutants, the AAC codon for Asn-100 of thebenA β-tubulin gene was altered to ATC, coding for Ile. Sequence displacement experiments confirmed that the substitution of Ile for Asn-100 confers resistance to rhizoxin in this organism. The amino acid sequences of β-tubulin surrounding the 100th amino acid residue from the N-terminus including Asn-100 are highly conserved with a few exceptions. The fission yeastSchizosaccharomyces pombe and the budding yeastSaccharomyces cerevisiae are naturally occurring Rhir organisms whose β-tubulin genes encode Ile and Val respectively at the 100th amino acid residue. The Ile-100 ofS. pombe and the Val-100 ofS. cerevisiae were altered to Asn using site-directed mutagenesis and gene displacement techniques. The resultant haploid strains of these two yeasts uniquely expressing β-tubulin (Asn-100) instead of β-tubulin (Ile-100 or Val-100) were found to be Rhis. Haploid yeast expressing β-tubulin (Asn-100) is normal except for its sensitivity to rhizoxin. These results suggest that rhizoxin resistance has a common basis in both naturally occurring species and experimentally selected mutants in the substitution of Ile or Val for Asn-100 in β-tubulin.
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  • 79
    ISSN: 1617-4623
    Keywords: Rhizopus aspartic proteinase ; Splicing of intron ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The intron of the Rhizopus aspartic proteinase gene (RNAP-I) was modified by in vitro mutagenesis and examined for its splicing efficiency in Saccharomyces cerevisiae. The wild-type intron of the RNAP-I gene was not spliced at all in spite of its structural similarity to introns of S. cerevisiae. The primary transcript of the RNAP-I gene was converted to correctly translatable mRNA only when the complete consensus sequence of S. cerevisiae introns (i.e. 5″-GTATGT-----TACTAAC-----TAG-3″) was introduced into its intron, although the efficiency of splicing was low. It is also shown that transformants carrying the RNAP-I gene with the complete consensus sequence of S. cerevisiae introns produce active RNAP-I protein.
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  • 80
    ISSN: 1617-4623
    Keywords: Cochliobolus heterostrophus race T disease ; Maize cytoplasmic male sterility ; Mitochondria ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We asked whether the mitochondrial T-urf13 gene, associated with the male sterility phenotype of T cytoplasm in maize, can be expressed in Saccharomyces cerevisiae and whether this expression can mimic the effects observed in maize. We introduced the universal code equivalent of the T-urf13 gene into the S. cerevisiae nucleus by transformation and directed its translation product into mitochondria by means of a fusion with the targeting presequence from Neurospora crassa ATPase subunit 9. We show that expression of the universal code equivalent of the T-urf13 gene in the yeast nucleus does indeed mimic its effects in maize: respiratory growth of yeast is inhibited, respiration-deficient cytoplasmic mutants accumulate and NADH oxidation of isolated mitochondria is uncoupled. All these effects are observed only if the mitochondrial targeting peptide and methomyl or HmT toxin are present.
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  • 81
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    Molecular genetics and genomics 223 (1990), S. 49-57 
    ISSN: 1617-4623
    Keywords: Neurospora crassa ; Heterologous gene expression ; Saccharomyces cerevisiae ; Domain independence ; TRP1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The trifunctional TRP1 gene from Neurospora crassa (N-TRP1) was subcloned into the yeast-Escherichia coli shuttle vector YEp13 and expressed in Saccharomyces cerevisiae. The three activities of the N-TRP1 gene product were detected in yeast mutants that lacked either N-(5′-phosphoribosyl) anthranilate (PRA) isomerase or both the glutamine amidotransferase function of anthranilate synthase and indole-3-glycerol phosphate (InGP) synthase. The protein was detected on immunoblots only as the full length 83 kda product indicating that the trifunctional gene product was expressed in yeast primarily in a fully active, undegraded form. By placing the subcloned N-TRP1 gene under the control of the inducible PHO5 promoter from yeast, the expression of all three activities was increased to more than ten fold that of wild-type yeast and the overproduced protein could be visualized by SDS-polyacrylamide gel electrophoresis of crude extract and Coomassie Blue staining. Using the expression system described the effect of selective deletion of regions of the coding sequence of the N-TRP1 gene on expression of the three activities was tested. Expression of either the F- or C-domains, catalyzing respectively the PRA isomerase or InGP synthase activities, did not depend on the presence of the other domain in the active polypeptide. Furthermore, normal dimer formation occurred with a protein active for InGP synthase in a deletion derivative lacking most of the PRA isomerase domain, ruling out the hypothesis that interaction between the active site regions for PRA isomerase and InGP synthase accounted for dimer formation in the trifunctional product.
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  • 82
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    Molecular genetics and genomics 223 (1990), S. 97-106 
    ISSN: 1617-4623
    Keywords: Heat shock ; Saccharomyces cerevisiae ; Stationary phase ; cAMP ; Thermotolerance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have isolated a new small heat shock gene, HSP12, from Saccharomyces cerevisiae. It encodes a polypeptide of predicted Mr 12 kDa, with structural similarity to other small heat shock proteins. HSP12 gene expression is induced several hundred-fold by heat shock and on entry into stationary phase. HSP12 mRNA is undetectable during exponential growth in rich medium, but low levels are present when cells are grown in minimal medium. Analysis of HSP12 expression in mutants affected in cAMP-dependent protein phosphorylation suggests that the gene is regulated by cAMP as well as heat shock. A disruption of the HSP12 coding region results in the loss of an abundant 14.4 kDa protein present in heat shocked and stationary phase cells. It also leads to the induction of the heat shock response under conditions normally associated with low-level HSP12 expression. The HSP12 disruption has no observable effect on growth at various temperatures, nor on the ability to acquire thermotolerance.
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  • 83
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    Molecular genetics and genomics 223 (1990), S. 342-344 
    ISSN: 1617-4623
    Keywords: rRNA genes ; Growth rate ; Yeast ; Pulsed field gel electrophoresis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary By employing pulsed field gel electrophoresis we find that slow growing strains of Kluyveromyces lactis have only 43%–55% of the wild-type level of ribosomal DNA (rDNA) repeats. When subjected to prolonged vegetative growth these strains can increase both the number of rDNA repeats and their growth rate.
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  • 84
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    Molecular genetics and genomics 223 (1990), S. 394-400 
    ISSN: 1617-4623
    Keywords: Yeast ; Mitochondria ; CBS2 antibodies ; CBS2 protein ; In vitro import
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The nuclear genome encoded yeast protein CBS2 is required for translational activation of mitochondrial cytochrome b RNA. Genetic studies have shown that the target sequence of the CBS2 protein is the 5′ untranslated leader sequence of cytochrome b RNA. Here we report on the intracellular localization of CBS2. CBS2 protein, expressed in Escherichia coli and prepared from inclusion bodies, was used as an antigen to raise a polyclonal rabbit antiserum. Affinity-purified CBS2 antibodies detect a 45 kDa protein in mitochondrial lysates of wild-type cells, which is absent in a strain in which the CBS2 gene has been deleted. The protein is overexpressed in mitochondrial extracts of a transformant carrying the CBS2 gene on a high copy number plasmid, but undetectable in the post-mitochondrial supernatant. Intramitochondrial localization of CBS2 was verified by in vitro import of CBS2 protein that had been synthesized in a reticulocyte lysate programmed with CBS2 mRNA transcribed in vitro. Mitochondrial import of CBS2 is not accompanied by any detectable proteolytic processing.
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  • 85
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    Molecular genetics and genomics 223 (1990), S. 474-480 
    ISSN: 1617-4623
    Keywords: Yeast ; Promoter ; Argininosuccinate lyase ; ARG4
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Transcription of the ARG4 gene of Saccharomyces cerevisiae is regulated by general control of amino acid biosynthesis but not by a specific regulatory mechanism. Three deletion mutants (ΔI, ΔII,, ΔIII) successively removing DNA sequences upstream from the coding sequence have been phenotypically analyzed after insertion into a single copy plasmid. As expected, ΔI, which lacks the sequences upstream to −155, including the two putative upstream activation sequences (UAS), was unable to derepress argininosuccinate lyase biosynthesis under conditions of amino acid starvation. In ΔII (deleted up to −126) the enzyme activity was very low and cells harbouring this allele were arginine dependent. These drastic phenotypic changes can be attributed to the loss of 12 out of 14 dA residues from positions −124 to −137. This poly (dAdT) sequence most likely serves as an upstream promoter element for constitutive expression of ARG4. The ΔIII deletion removes all 5′ sequences including the putative TATA box. This inactive allele has been successfully used for selecting yeast promoters of unknown origin.
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  • 86
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    Molecular genetics and genomics 221 (1990), S. 353-357 
    ISSN: 1617-4623
    Keywords: Yeast ; DNA alkylation ; DNA repair
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary Repair of methylated bases in Saccharomyces cerevisiae was measured by two methods: in vitro in cell extracts, and in vivo, by determining the loss of methylated bases from yeast DNA after treatment of stationary cultures with [3H]-N-methyl-N′-nitro-N-nitrosoguanidine. Whereas no repair activity could be detected by the in vitro method, the methylated bases were removed in vivo very efficiently. These contradictory results of in vitro and in vivo repair measurements suggest that either the repair enzymes of yeast are sufficiently different from those of bacteria and mammalian cells that they are not active in the in vitro assay, or that methylated bases are repaired in yeast by a different pathway.
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  • 87
    ISSN: 1617-4623
    Keywords: Intrachromosomal and interchromosomal recombination ; Saccharomyces cerevisiae ; RAD18
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The frequency of intra- and interchromosomal recombination was determined in RAD18 and rad18 deletion and rad18-3 mutant strains. It was found that spontaneous interchromosomal recombination at trp5, his1, ade2, and MAT was elevated 10- to 70-fold in the rad18-3 and rad18Δ mutants as compared to the RAD + strains. On the other hand the frequencies of spontaneous intrachromosomal recombination for the his3Δ3′, his3Δ5′ and the his4C −, his4A − duplications and for heterothallic mating type switching were only marginally elevated in the rad18 deletion mutant, and recombination between ribosomal DNA repeats was only 2-fold elevated in the rad18-3 mutant. These differences may be due to a haploid versus diploid specific difference. However interchromosomal recombination was elevated 40-fold and intrachromosomal recombination was only marginally (1.5-fold) elevated in a diploid homozygous for rad18Δ, arguing against a haploid versus diploid specific difference. Possible explanations for the difference in the elevated levels of intra- versus interchromosomal spontaneous recombination are discussed.
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  • 88
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Schizosaccharomyces pombe ; Transcription initiation ; ADH gene ; TATA sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The roles of the TATA element and sequences near the mRNA initiation site in specifying the location of initiation sites in Saccharomyces cerevisiae were examined, using the Schizosaccharomyces pombe ADH gene. The importance of spacing was demonstrated by analysis of a series of deletions that removed from 8–50 bp between the TATA element and ATG translation initiation site of this gene. Primer extension mapping showed that increasing deletion length is associated with a progressive shift downstream in the location of the initiation sites. The distance of a given site from the promoter affected the relative ability of the site to be utilized for initiation. For this gene, a permissive region for transcription initiation exists between 55 and 125 bases downstream of the TATA element, and a zone of 75–115 bases allows maximal usage of an initiation site. The presence of a TATA sequence was shown to be necessary in S. cerevisiae for maintaining the location of this “window” of initiation. The TATA sequence is essential for function of the gene in S. pombe. This gene, as well as the majority of the 63 S. cerevisiae genes surveyed, uses initiation sites which fit a PyAA/T(Pu) consensus. Cis-acting mutations were recovered which restored ADH activity to a deletion allele that initiates its mRNAs downstream of the ATG. DNA sequence and transcript analysis with these mutants confirmed the requirement of proper spacing and conformity of initiation sites to the PyAA/T(Pu) consensus for efficient transcript initiation.
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  • 89
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    Molecular genetics and genomics 223 (1990), S. 438-442 
    ISSN: 1617-4623
    Keywords: Transcription ; Promoter ; Oligo(dA-dT) stretch ; Gel shift assays ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Oligo(dA-dT) tracts are frequently found in the intergenic regions of the yeast Saccharomyces cerevisiae and have been proposed to act as upstream promoter elements for constitutive transcription. An oligo(dA-dT) tract of 23 bp is also found as a characteristic sequence motif in the centre of the 230 by segment which separates the open reading frames of the CBS2 gene and its 5′-flanking gene on chromosome IV. Recently we have reported that transcription of CBS2 is initiated immediately adjacent to this oligo(dA-dT) tract (Michaelis et al. 1988). Here we report that the flanking gene of unknown function is divergently transcribed into an RNA with heterogeneous 5′ ends. Two of these 5′ ends map within the oligo(dA-dT) stretch, while the third is located upstream, leading to an RNA species which is partially complementary to the CBS2 transcript. Gel shift assays show that the oligo(dA-dT) stretch is specifically recognized by (a) binding factor(s) in nuclear extracts. We discuss these results with respect to the role of oligo(dA-dT) stretches in gene expression in yeast.
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  • 90
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Gene regulation ; TUF ; Pyruvate decarboxylase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The upstream activation site of the pyruvate decarboxylase gene, PDC1, of Saccharomyces cerevisiae contains an RPG box, and mediates the increase in expression of a PDC1-lacZ fusion gene during growth on glucose. Oligonucleotide replacement experiments indicate that the RPG box functions as an absolute activator of expression, but other elements (possibly CTTCC repeats) are required for carbon source regulation, and maximal expression. Gel retardation and oligonucleotide competition experiments suggest that the DNA binding factor TUF interacts with the RPG box in the upstream region of PDC1. Binding of TUF factor is not carbon source dependent in in vitro experiments, and is probably not responsible for glucose induction of PDC1 expression.
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  • 91
    ISSN: 1573-5036
    Keywords: Gaeumannomyces graminis ; genotypes ; interaction ; manganese ; oxidation ; take-all ; Triticum aestivum ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Take-all is a world-wide root-rotting disease of cereals. The causal organism of take-all of wheat is the soil-borne fungus Gaeumannomyces graminis var tritici (Ggt). No resistance to take-all, worthy of inclusion in a plant breeding programme, has been discovered in wheat but the severity of take-all is increased in host plants whose tissues are deficient for manganese (Mn). Take-all of wheat will be decreased by all techniques which lift Mn concentrations in shoots and roots of Mn-deficient hosts to adequate levels. Wheat seedlings were grown in a Mn-deficient calcareous sand in small pots and inoculated with four field isolates of Ggt. Infection by three virulent isolates was increased under conditions which were Mn deficient for the wheat host but infection by a weakly virulent isolate, already low, was further decreased. Only the three virulent isolates caused visible oxidation of Mn in vitro. The sensitivity of Ggt isolates to manganous ions in vitro did not explain the extent of infection they caused on wheat hosts. In a similar experiment four Australian wheat genotypes were grown in the same Mn-deficient calcareous sand and inoculated with one virulent isolate of Ggt. Two genotypes were inefficient at taking up manganese and were very susceptible to take-all, one was very efficient at taking up manganese and was resistant to take-all, and the fourth genotype was intermediate for both characters. All genotypes were equally resistant under Mn-adequate conditions.
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  • 92
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    Euphytica 45 (1990), S. 71-80 
    ISSN: 1573-5060
    Keywords: wheat ; Triticum aestivum ; wheat leaf rust ; Puccinia recondita f.sp. tritici ; partial resistance ; histology ; growth curve
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The average size of wheat leaf rust colonies, measured using epifluorescence microscopy was significantly larger in the highly susceptible genotype Morocco than in the susceptible genotype Kaspar and the partially resistant genotypes Westphal 12A, Akabozu and BH 1146. This was already so three days after inoculation. Colony growth in partially resistant genotypes was continuously retarded compared to colonies in the highly susceptible genotype Morocco. No evidence was found for an initial inhibition of the growth of colonies in partially resistant genotypes. In partially resistant genotypes formation of uredial beds and sporulating areas started at a smaller colony size than in susceptible genotypes. Wheat leaf rust colonies in primary leaves of all genotypes studied were much larger than colonies in flag leaves measured at the same number of days after inoculation. Growth and sporulation of not intertwined colonies was not influenced by either a high or a low number of neighbouring colonies.
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  • 93
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    Euphytica 45 (1990), S. 81-86 
    ISSN: 1573-5060
    Keywords: Triticum aestivum ; wheat ; Puccinia recondita f.sp. tritici ; wheat leaf rust ; partial resistance ; histology ; abortion ; adult plant resistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Arrest of the growth of wheat leaf rust infection structures was studied with fluorescence microscopy in seedling leaves and flag leaves of the susceptible spring wheat genotypes Morocco and Kaspar and the partially resistant genotypes Westphal 12A and Akabozu. The percentages non-penetrants and substomatal vesicle abortion were low in all genotypes. In the partially resistant genotypes the percentage abortion of infection structures was higher than in the susceptible genotype Morocco. Aborted infection structures had formed one or two haustorial mother cells. In adult plants differences in the percentage aborted infection structures between susceptible and partially resistant genotypes were more pronounced than in seedlings. The so-called late abortion was not observed.
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  • 94
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    Euphytica 45 (1990), S. 59-69 
    ISSN: 1573-5060
    Keywords: Triticum aestivum ; wheat ; Pyrenophora tritici-repentis ; resistance ; tan spot ; yellow spot ; variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary No complete resistance to Pyrenophora tritici-repentis has been located in more than 1400 bread wheats examined. Incomplete resistance, however, occurs at different levels in many spring and winter types and data are presented for the strongest sources of resistance detected. In particular, there is a high frequency of Brazilian spring wheats with appreciable levels of resistance to this pathogen. Recent international nurseries from CIMMYT, Mexico, also contain numerous potentially valuable sources of resistance and these wheats may be shorter and higher yielding in Australia than the Brazilian wheats. The resistances in many Brazilian cultivars may be largely common because the cultivars are often strongly interrelated. Some of the Brazilian wheats resistant to P. tritici-repentis are also resistant to one or more of the septoria diseases and/or possess tolerance to aluminium toxicity.
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  • 95
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    Euphytica 45 (1990), S. 87-92 
    ISSN: 1573-5060
    Keywords: Triticum aestivum ; wheat ; near-isogenic lines ; NILs ; Puccinia recondita f.sp. tritici ; leaf rust ; Puccinia striiformis ; yellow rust ; backcross ; variation ; background resistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Using the cultivar Arina as the recurrent parent, six backcrosses were made with two donor lines carrying the leaf rust resistance genes Lr1 and Lr9, respectively. Selection for leaf rust resistance occurred at the seedling stage in the greenhouse; the first plants transferred to the field were BC6F4s. Frequency distribution of the 332 Lr1/7 × Arina and the 335 Lr9/7 × Arina lines showed continuous variation for yellow rust resistance and heading date in these leaf rust near-isogenic lines (NILs). Similar results were also obtained for plant height, for resistance to powdery mildew and glume blotch, as well as for baking quality characters in another set of more advanced NILs. The available information on the behaviour of one of the parents of cultivar Arina led to the conclusion that the expressed yellow rust resistance is quantitative and might possibly be durable.
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  • 96
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    Euphytica 45 (1990), S. 169-177 
    ISSN: 1573-5060
    Keywords: Triticum aestivum ; wheat ; coefficient of parentage ; pedigree ; gene pool
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Pedigrees of 142 Yugoslavian winter wheat cultivars were traced to 110 ancestral genotypes, of which 41 contributed significantly. In each of the four major Yugoslavian wheat breeding programs, the most important ancestor, as evaluated by mean coefficient of parentage, was ‘Akagomughi’, source of the genes Rht8 and Ppd1. The other 13–19 ancestors accounting for the majority of the remaining germplasm, varied considerably among institutions. The relative contributions of ancestors changed little between the periods 1967–76 and 1982–86, with the exception of ‘Neuzucht’ (source of a 1B/1R translocation), which became much more important in the latter period.
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  • 97
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    Euphytica 46 (1990), S. 149-155 
    ISSN: 1573-5060
    Keywords: Triticum aestivum ; wheat ; selection ; preharvest sprouting ; germination ; kernel color ; dormancy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The utility of spike- and seed-based mass selection techniques for improving preharvest sprouting resistance in heterogeneous wheat (Triticum spp.) populations was evaluated. Sorting seed by size improved selection efficiency in some cases, putatively by physiological synchronization. Progeny testing, as well as changes in frequency of red-kernelled types, indicate effectiveness of both spike- and seed-based mass selection for reduced preharvest sprouting. Differential effectiveness of mass selection, in populations segregating for dormancy from different sources, is consistent with previous work on mechanisms of dormancy from these sources. These results are of value to improvement of preharvest sprouting resistance in large, heterogeneous wheat populations.
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  • 98
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    Euphytica 47 (1990), S. 165-169 
    ISSN: 1573-5060
    Keywords: Triticum aestivum ; wheat ; Secale cereale ; rye ; isozyme loci ; esterase ; homoeology relationships
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The EST-6 leaf esterase phenotypes from euploid, nullisomic-tetrasomic and rye chromosome addition and substitution lines of common wheat were determined using polyacrylamide gel electrophoresis. Evidence is presented to demonstrate that Est-6 is a new set of genes, that are expressed in the leaf. The Est-6 gene set were clearly distinguished from the Est-5 genes which are expressed in the grain. The three homoeoallelic loci, Est-A6, Est-B6 and Est-D6, were located on chromosomes 3A, 3B and 3D. An Est-R6 gene was located on chromosome 6R is involved in rye. Some considerations concerning homoeology between homoeologous group 3 of wheat and the rye chromosome 6R are made.
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  • 99
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    Euphytica 48 (1990), S. 1-8 
    ISSN: 1573-5060
    Keywords: Triticum aestivum ; wheat ; protein accumulation ; plant protein ; protein estimation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Better understanding of the physiological and genetic basis of wheat grain protein will contribute to breeding efforts for this characteristic. This study provides information about plant protein distribution in high and low grain protein winter wheats (Triticum aestivum L.) at different growth stages and its relation to grain protein. Field experiments involved two winter wheats with high grain protein, ‘Redwin’ and ‘Lancota’, and two with low grain protein, ‘Centurk’ and ‘Brule’ in two years. Protein content in the head, the upper three leaves, the first and second leaf, and the peduncle were estimated with Near Infrared Reflectance Spectrophotometer (NIR) at five growth stages. High protein cultivars had higher leaf protein at ripe and higher protein content in the heads at most growth stages than low grain protein cultivars. High protein cultivars had lower protein content in the peduncle than low protein cultivars at ripe. Correlation coefficients between plant-part protein and grain protein ranged from 0.48 to 0.87 for the heads, from −0.45 to −0.79 for the peduncle, and from 0.55 to 0.84 for the leaves. A combination of head, peduncle, and first leaf protein at heading was significantly related to grain protein (R2=0.71). Indirect selection for head, peduncle, and first leaf (flag leaf) protein at heading should result in increased grain protein. Recurrent selection for increased grain protein, with parent selectionbefore anthesis and hybridization should be successful.
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  • 100
    ISSN: 1573-5060
    Keywords: Sorghum bicolor ; sorghum ; Triticum aestivum ; wheat ; Oryza sativa ; rice ; Fraction-1-Protein inheritance ; Isoelectric focusing ; intergeneric hybrids ; Large and small sub-units ; rice × sorghum ; rice × wheat hybrids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The polypeptide composition of Fraction-1-Protein (F1P) from rice × sorghum, rice × wheat hybrids and their respective parents have been analyzed by a microelectrofocusing method. The large sub-unit (LSU) is composed of three polypeptides and the small sub-unit (SSU) of two polypeptides in rice and sorghum parents and rice × sorghum hybrids. Similarly, LSU is composed of three polypeptides in the rice and wheat parents and rice × wheat hybrids. Two polypeptides occur in the SSU of rice parent and rice × wheat hybrids where as only one polypeptide in the wheat parent. These polypeptides also differ in their isoelectric points. Based on the previous reports of F1P inheritance in hybrids in other crops, F1P analysis of rice × sorghum and rice × wheat hybrids does not seem to be an important marker to identify such intergeneric hybrids. Since this is first such report of F1P inheritance in hybrids between distantly related plants, its implication in different modes of inheritance are discussed.
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