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  • Saccharomyces cerevisiae
  • gene expression
  • Springer  (65)
  • Wiley-Blackwell  (20)
  • 2020-2024
  • 1990-1994  (85)
  • 1990  (85)
  • 1
    Electronic Resource
    Electronic Resource
    Developmental changes of aldehyde dehydrogenase isozymes in human livers: Mitochondrial ALDH2 isozyme is expressed in fetal livers (1990)
    Springer
    Cellular and molecular life sciences 46 (1990), S. 747-750 
    Details
    ISSN: 1420-9071
    Keywords: Aldehyde dehydrogenase ; developmental changes ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Previous reports suggested that the major cytosolic aldehyde dehydrogenase (ALDH1) was present in fetal and infant livers, but the major mitochondrial isozyme (ALDH2) was absent or severely diminished. Re-examination by means of starch gel electrophoresis followed by enzyme activity staining, and by means of dot blot immuno-hybridization of liver samples with known genotypes of theALDH 2 locus, indicated that bothALDH 1 andALDH 2 genes are expressed in fetal and infant livers. In addition, ALDH4 isozyme was also observed. The results imply that a fetus with the ‘usual’ homozygousALDH 2 1 /ALDH 2 1 genotype, but not one with the atypicalALDH 2 1 /ALDH 2 2 orALDH 2 2 /ALDH 2 2 , is capable of detoxifying acetaldehyde transferred from the mother.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01939955
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  • 2
    Electronic Resource
    Electronic Resource
    The OGD1 gene, affecting 2-oxoglutarate dehydrogenase in S. cerevisiae, is closely linked to HIS5 on chromosome IX (1990)
    Springer
    Current genetics 17 (1990), S. 85-88 
    Details
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00313254
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  • 3
    Electronic Resource
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    Cloning and sequencing of URA10, a second gene encoding orotate phosphoribosyl transferase in Saccharomyces cerevisiae (1990)
    Springer
    Current genetics 17 (1990), S. 105-111 
    Details
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00312853
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  • 4
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    Isolation and properties of yeast mutants affected in farnesyl diphosphate synthetase (1990)
    Springer
    Current genetics 18 (1990), S. 41-46 
    Details
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00321113
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  • 5
    Electronic Resource
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    Expression of the Aspergillus niger glucose oxidase gene in A. niger, A. nidulans and Saccharomyces cerevisiae (1990)
    Springer
    Current genetics 18 (1990), S. 531-536 
    Details
    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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    URL: http://dx.doi.org/10.1007/BF00327024
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  • 6
    Electronic Resource
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    Effect of chromosome loss on the leavening ability of Saccharomyces cerevisiae in dough (1990)
    Springer
    Current genetics 18 (1990), S. 401-403 
    Details
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00309908
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  • 7
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    Isolation and characterization of the Pichia stipitis xylitol dehydrogenase gene, XYL2, and construction of a xylose-utilizing Saccharomyces cerevisiae transformant (1990)
    Springer
    Current genetics 18 (1990), S. 493-500 
    Details
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00327019
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  • 8
    Electronic Resource
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    A recessive mutant allele of the HNM1 gene of Saccharomyces cerevisiae is responsible for hyper-resistance to nitrogen mustard (1990)
    Springer
    Current genetics 18 (1990), S. 187-192 
    Details
    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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    URL: http://dx.doi.org/10.1007/BF00318378
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  • 9
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    G418-resistance as a dominant marker and reporter for gene expression in Saccharomyces cerevisiae (1990)
    Springer
    Current genetics 18 (1990), S. 303-313 
    Details
    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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    URL: http://dx.doi.org/10.1007/BF00318211
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  • 10
    Electronic Resource
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    When a glycolytic gene on a yeast 2μORI-STB plasmid is made essential for growth its expression level is a major determinant of plasmid copy number (1990)
    Springer
    Current genetics 17 (1990), S. 119-123 
    Details
    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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    URL: http://dx.doi.org/10.1007/BF00312855
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  • 11
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    Functional analysis of the sporulation-specific SPR6 gene of Saccharomyces cerevisiae (1990)
    Springer
    Current genetics 18 (1990), S. 293-301 
    Details
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00318210
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  • 12
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    Interactions between the yeast mitochondrial and nuclear genomes: isogenic suppressive and hypersuppressive petites differ in their resistance to the alkaloid lycorine (1990)
    Springer
    Current genetics 17 (1990), S. 455-457 
    Details
    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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    URL: http://dx.doi.org/10.1007/BF00334527
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  • 13
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    Evolutionary conservation of transcriptional machinery between yeast and plants as shown by the efficient expression from the CaMV 35S promoter and 35S terminator (1990)
    Springer
    Current genetics 17 (1990), S. 473-479 
    Details
    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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    URL: http://dx.doi.org/10.1007/BF00313074
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  • 14
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    The absence of introns in yeast mitochondria does not abolish mitochondrial recombination (1990)
    Springer
    Current genetics 17 (1990), S. 537-541 
    Details
    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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    URL: http://dx.doi.org/10.1007/BF00313085
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  • 15
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    Fate of highly expressed proteins destined to peroxisomes in Saccharomyces cerevisiae (1990)
    Springer
    Current genetics 18 (1990), S. 23-27 
    Details
    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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    URL: http://dx.doi.org/10.1007/BF00321111
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  • 16
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    Effect of nystatin, amphotericin B and amphotericin B methyl ester on Saccharomyces cerevisiae with different lipid composition (1990)
    Springer
    Mycopathologia 112 (1990), S. 165-172 
    Details
    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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    URL: http://dx.doi.org/10.1007/BF00436649
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  • 17
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    Determination of the role of polyphosphate in transport-coupled phosphorylation in the yeast Saccharomyces cerevisiae (1990)
    Springer
    Antonie van Leeuwenhoek 57 (1990), S. 159-164 
    Details
    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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    URL: http://dx.doi.org/10.1007/BF00403950
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  • 18
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    Genetic and molecular approaches to synthesis and action of the yeast killer toxin (1990)
    Springer
    Cellular and molecular life sciences 46 (1990), S. 193-200 
    Details
    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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    URL: http://dx.doi.org/10.1007/BF02027313
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  • 19
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    Purification and characterization of a (R)-2,3-butanediol dehydrogenase from Saccharomyces cerevisiae (1990)
    Springer
    Archives of microbiology 154 (1990), S. 267-273 
    Details
    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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    URL: http://dx.doi.org/10.1007/BF00248966
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  • 20
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    Absence of glucose-induced cAMP signaling in the Saccharomyces cerevisiae mutants cat1 and cat3 which are deficient in derepression of glucose-repressible proteins (1990)
    Springer
    Archives of microbiology 154 (1990), S. 199-205 
    Details
    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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    URL: http://dx.doi.org/10.1007/BF00423333
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  • 21
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    Growth of catalase A and catalase T deficient mutant strains of Saccharomyces cerevisiae on ethanol and oleic acid (1990)
    Springer
    Archives of microbiology 153 (1990), S. 513-517 
    Details
    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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    Effects of ethanol on Saccharomyces cerevisiae as monitored by in vivo 31P and 13C nuclear magnetic resonance (1990)
    Springer
    Archives of microbiology 153 (1990), S. 384-391 
    Details
    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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    Genetic mapping and biochemical analysis of mutants in the maltose regulatory gene of the MAL1 locus of Saccharomyces cerevisiae (1990)
    Springer
    Archives of microbiology 154 (1990), S. 544-549 
    Details
    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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    The effect of insertion of the maize transposable elementMutator is dependent on genetic background (1990)
    Springer
    Biochemical genetics 28 (1990), S. 9-20 
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    ISSN: 1573-4927
    Keywords: mutator ; transposable element ; alcohol dehydrogenase ; maize ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A secondary mutant, derived from an allele of maize alcohol dehydrogenase 1 (Adh1) carrying a Mutator transposable element (Mu1) in its first intron, was reported to exhibit a threefold decrease in ADH enzymatic activity and steady-state RNA levels compared to the original mutant. The original mutant,Adh1-S3034 (abbreviatedS3034), was previously characterized at the molecular level. The derivative, abbreviatedS3034b, has now been cloned; at the DNA sequence level the insertion and surroundingAdh1 sequences are indistinguishable fromS3034. Furthermore, in our lines there is no difference in relative ADH activities between products of the two putative alleles. A comparison of gene expression in heterozygotes obtained by crossing to different tester lines reveals a correlation between the measured decrease in levels of ADH polypeptide produced by the mutant allele and the background in which it is measured; this effect is distinct from any background-related variation in the expression of the progenitor allele. It does not appear to be attributable to alternative patterns of DNA modification. It appears to reflect a background-associated difference in the level of normalAdh1-RNA produced. Thus the previously reported distinction betweenS3034 andS3034b may be due to differences in the extent to which the mutant allele and a given genetic background interact to produce functionalAdh1-RNA.
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    Expression of a functionally active cardiac fatty acid-binding protein in the yeast, Saccharomyces cerevisiae (1990)
    Springer
    Molecular and cellular biochemistry 98 (1990), S. 69-74 
    Details
    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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    The gene coding for glutamate dehydrogenase inDrosophila melanogaster (1990)
    Springer
    Biochemical genetics 28 (1990), S. 337-346 
    Details
    ISSN: 1573-4927
    Keywords: glutamate dehydrogenase ; Drosophila melanogaster ; gene expression ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract We have isolated theDrosophila melanogaster locus coding forl-glutamate dehydrogenase (EC 1.4.1.3) by virtue of its similarity to the corresponding human gene. There is only one copy of this gene in the fruit fly genome, located on the right arm of chromosome 3 (95D1-4). The transcript includes at least one large intron and matures to a ∼2.4-kb-long polyadenylated RNA whose expression is under developmental control.
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    The gene coding for glutamate dehydrogenase inDrosophila melanogaster (1990)
    Springer
    Biochemical genetics 28 (1990), S. 337-346 
    Details
    ISSN: 1573-4927
    Keywords: glutamate dehydrogenase ; Drosophila melanogaster ; gene expression ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract We have isolated theDrosophila melanogaster locus coding forl-glutamate dehydrogenase (EC 1.4.1.3) by virtue of its similarity to the corresponding human gene. There is only one copy of this gene in the fruit fly genome, located on the right arm of chromosome 3 (95D1-4). The transcript includes at least one large intron and matures to a ∼2.4-kb-long polyadenylated RNA whose expression is under developmental control.
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    URL: http://dx.doi.org/10.1007/BF02401423
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    Changes in alpha-globin gene expression in mice of two alpha-globin haplotypes during development (1990)
    Springer
    Biochemical genetics 28 (1990), S. 445-457 
    Details
    ISSN: 1573-4927
    Keywords: mouse ; hemoglobin ; embryonic ; adult ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Adult alpha-globin in mice is synthesized in large amounts during development, first in the primitive, nucleated erythrocytes of yolk sac origin and later in the definitive, nonnucleated erythrocytes that differentiate in the fetal liver, spleen, and bone marrow. Isoelectric focusing analysis of hemoglobins of mice with theHba g2 andHba c haplotypes shows that the ratios of alpha chain 1 to chain 5m and alpha chain 1 to chain 4 in adult hemoglobins fromHba g2 andHba c mice, respectively, change between day 11.5 and day 16.5 of gestation in nucleated red cells, while no change occurs in nonnucleated red cells. The percentage ratios of the two different alpha-globin chains are different inHba g2 andHba c mice for EII, EIII, and adult hemoglobin. In nucleated red cells of yolk sac origin, differences and changes in alpha-globin ratios are a composite of changing globin gene transcription and posttranslational competitive affinities among globins to form embryonic and adult hemoglobin tetramers.
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    An altered constitutive peptide in sym 5 mutants of Pisum sativum L. (1990)
    Springer
    Plant molecular biology 14 (1990), S. 207-216 
    Details
    ISSN: 1573-5028
    Keywords: developmental mutant ; gene expression ; nodule formation ; Pisum sativum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mutational analysis of Pisum sativum L. was used to search for constitutive proteins that might function in nodule formation. The sym 5 locus is a mutational hot spot, represented by seven independently derived mutant lines with decreased nodulation. Comparison of two-dimensional polyacrylamide gels of in vitro-translated root RNA showed a consistent difference in the migrational pattern of one peptide. In the nodulating parental cultivar ‘Sparkle’, a 66 kDa peptide had a pI of 5.9. In four of the five tested sym 5 mutants, the 66 kDa peptide had a more acidic pI of 5.8. This 66 kDa peptide is found in lateral root, tap root, and shoot. Its expression was independent of rhizobial inoculation, root temperature, or light.
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    Molecular cloning and sequencing of a cDNA for an auxin-repressed mRNA: correlation between fruit growth and repression of the auxin-regulated gene (1990)
    Springer
    Plant molecular biology 14 (1990), S. 127-136 
    Details
    ISSN: 1573-5028
    Keywords: auxin action ; cDNA clone ; gene expression ; developmental regulation ; strawberry fruit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A complementary DNA (cDNA) library has been constructed in λgt10 from poly(A)+ mRNA isolated from auxin-deprived strawberry receptacles. By differential plaque filter hybridization, a cDNA (λSAR5) to an auxin-repressed mRNA has been isolated. The expression of the auxin-repressed gene is studied at various stages of normal fruit development and in fruits of variant strawberry genotype using λSAR5 as a probe. Northern analyses of RNA isolated from pollinated and unpollinated fruits of various developmental stages revealed that mRNA corresponding to the λSAR5 clone is repressed during normal fruit development, and the level of λSAR5 mRNA is regulated by endogenous auxin. Furthermore, results with both normal and variant genotype strawberry fruit indicate that there is a positive correlation between growth of strawberry fruit and repression of mRNA corresponding to the λSAR5 clone. The λSAR5 cDNA has been sequenced and is 723 nucleotides in length. The deduced protein has 111 amino acid residues with a molecular mass of 12.5 kDa. The putative polypeptide starts at nucleotide position 20 and ends at 352. The molecular weight of the predicted polypeptide is in agreement with the molecular weight of the in vitro translated polypeptide of hybrid selected mRNA. A comparison of the nucleotide and deduced amino acid sequence of λSAR5 with nucleotide and protein sequences in data banks has not revealed any homology to known proteins.
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    Molecular cloning of cDNAs for auxin-induced mRNAs and developmental expression of the auxin-inducible genes (1990)
    Springer
    Plant molecular biology 14 (1990), S. 643-653 
    Details
    ISSN: 1573-5028
    Keywords: auxin action ; cDNA clones ; developmental regulation ; gene expression ; strawberry fruit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract By differential hybridization, two auxin-inducible cDNA clones (λSAR1 and λSAR2) have been isolated from a cDNA library constructed to poly(A)+ mRNA from auxin-treated strawberry receptacles. Both the clones have been used as probes to study the expression of the auxin-induced genes in pollinated and unpollinated fruits of various stages of development and in different organs. A high level of auxin-induced mRNAs is found in pollinated fruits as compared to unpollinated fruits of the same age, suggesting that the expression of the auxin-induced genes is developmentally regulated and the level of auxin-induced mRNAs is regulated by endogenous auxin. Furthermore, our data on the expression of λSAR1 and λSAR2 genes in pollinated and unpollinated fruits revealed a positive correlation between growth of strawberry fruit and the induction of mRNA corresponding to the λSAR1 and λSAR2 clones. Ethylene has no effect on the expression of the auxin-induced mRNAs. λSAR1 mRNA is not detected in other parts of strawberry plants whereas λSAR2 mRNA is present in roots. Furthermore, mRNA corresponding to λSAR1 and λSAR2 is not detected in other auxin-responsive plant systems such as pea epicotyls and bean explants.
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    Molecular cloning of a lupin-specific gene from a cDNA library of suspension-cultured cells of Lupinus polyphyllus (1990)
    Springer
    Plant molecular biology 15 (1990), S. 175-176 
    Details
    ISSN: 1573-5028
    Keywords: Lupinus polyphyllus ; cell culture ; cDNA clone ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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    URL: http://dx.doi.org/10.1007/BF00017739
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    Prosomes, subcomplexes of untranslated mRNP (1990)
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    Molecular biology reports 14 (1990), S. 1-9 
    Details
    ISSN: 1573-4978
    Keywords: prosomes ; messenger RNP ; intermediate filaments ; heat shock complex ; multicatalytical protease (MCP) ; protein synthesis ; gene expression ; differentiation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract PROSOMES are a novel class of small RNP particles of uniform morphology, but of variable RNA (pRNA) and protein composition (about 650 000 MW; 12 nm diameter in the EM). They were discovered as subcomplexes of free mRNP, tightly attached to inactive mRNA in the cytoplasm. The pRNAs hybridize stably to mRNA. Prosomes associate in vitro to mRNA and inhibit cell free protein synthesis inducing an mRNA structure unable to interact with ribosomes. Many types of prosomes were observed. The individual particle is made up by a variable combination of about 20 characteristic proteins and one or several pRNA. Some prosomal proteins are glycosylated, phosphorylated and, possibly, ADP-ribosylated and are highly conserved in evolution whilst others vary with the species and the mRNA population they are associated to. A protease activity was found associated to prosomes. The function(s) of the prosomes is(are) still unknown. The differential inhibition of in vitro protein synthesis points to a capacity to recognize mRNA and to keep it in an inactive state. The observation with the aid of monoclonal antibodies (pMABs) that prosomes and thus mRNP are attached to the intermediate filaments (IF) raises the question if one of the functions of the IF might be in the topological distribution of mRNA within the cell. Similar to the cytokeratin fibers, the prosome networks bridge neighboring cells at specific positions. — The nucleus also contains some prosomal antigens, located on chromosomes and on the nuclear matrix. Their presence and distribution in the cell compartments varies with the cell type and the prosomal antigen probed. Oocytes contain large amounts of prosomes. In embryonic development, the synthesis of individual prosomal proteins starts progressively after the blastula stage and resumes fully in gastrulation only; cleavage and blastula stage prosomes are thus of maternal origin. The nucleo-cytoplasmic distribution of prosomal antigens changes in embryos, with the stage of development and type of differentiation. In human tissues specific patterns of prosomal antigens were found in function of cell type and differentiation. In view of these data, the hypothesis may be formulated that prosomes are a population of mRNA-linked RNP which includes particles of varying individual composition and hence specificity. Attached to IF sub-networks, specific types of prosomes might accompany families of mRNA in function of the physiological state and the specialisation of given differentiated cell types. The cell-type specific organisation of the IF networks might be related to the messenger RNA complement of a given cell, and to its status of gene expression. The prosomes might thus have a function in controlling the transport, distribution and control of activity of specific mRNAs in the cell.
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    Cytokinin enhancement of the light induction of nitrate reductase transcript levels in etiolated barley leaves (1990)
    Springer
    Plant molecular biology 14 (1990), S. 585-594 
    Details
    ISSN: 1573-5028
    Keywords: cytokinin ; gene expression ; mRNA ; nitrate reductase ; transcriptional regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To investigate the molecular mechanism of cytokinin regulation of nitrate reductase (NR) activity, the influence of benzyladenine (BA) on the level of NR transcript was studied in etiolated barley leaves using a barley NR cDNA as a probe. Northern blot analyses of the levels of NR poly (A)+ RNA indicate that the amount present is proportional to the concentration of BA (2×10-8 to 2×10-4 M) applied to the leaves. Enhancement of NR mRNA by 2×10-5 M BA was clearly detected after 15 minutes of exposure of the leaves to light. The enhancement is cytokinin-specific and adenine is ineffective. Brief treatment with the protein synthesis inhibitor, cycloheximide, inhibited BA-enhanced NR activity but did not inhibit BA-enhanced NR transcript level, thus the enhancement was independent of concurrent protein synthesis. Nuclear runoff transcription studies showed that the enhancement of NR mRNA was at least partially due to increased transcription rates.
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    Expression of glutamine synthetase genes in roots and nodules of Phaseolus vulgaris following changes in the ammonium supply and infection with various Rhizobium mutants (1990)
    Springer
    Plant molecular biology 14 (1990), S. 549-560 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; glutamine synthetase ; leghaemoglobin ; nitrogen metabolism ; Phaseolus vulgaris ; root nodules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In this paper we have examined whether the four glutamine synthetase (gln) genes, expressed in roots and nodules of Phaseolus vulgaris are substrate-inducible by ammonium. Manipulation of the ammonium pool in roots, through addition and removal of exogenous ammonium, did not elicit any changes in the abundances of the four mRNAs thus suggesting that the gln genes in roots of this legume are neither substrate-inducible by ammonium nor derepressed during nitrogen starvation. In nodules the effect of the ammonium supply on expression of the gln genes has been examined by growing nodules under argon/oxygen atmospheres, or with a number of Fix- Rhizobium mutants, and following addition of exogenous ammonium. The results of these experiments suggest that the expression of the gln-γ gene, which is strongly induced during nodule development, is primarily under a developmental control. However nitrogen fixation appears to have a quantitative effect on expression of gln-γ as the abundance of this mRNA is about 2 to 4-fold higher under nitrogen-fixing conditions. This effect could not be mimicked by addition of exogenous ammonium and moreover is not specific to the gln-γ gene as mRNA from a leghaemoglobin gene was similarly affected. Taken together these results have failed to find an effect of ammonium on specifically inducing the expression of glutamine synthetase genes in roots and nodules of P. vulgaris.
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    Organ-specific modulation of gene expression in transgenic plants using antisene RNA (1990)
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    Plant molecular biology 15 (1990), S. 39-47 
    Details
    ISSN: 1573-5028
    Keywords: antisense ; gene expression ; plants ; regulation ; RNA ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have shown leaf-specific inhibition GUS gene expression in transgenic Nicotiana plants using an antisense RNA with a 41-base homology spanning the translation start codon of the gene. GUS was expressed from the nominally constitutive 35S promoter and the antisense RNA was expressed from the light-regulated ca/b promoter of Arabidopsis thaliana. A range of GUS inhibition from 0 to 100% was obtained by screening a small population of transgenic plants and the specific levels of inhibition observed were stably inherited in two generations. An antiGUS ‘gene’ dosage effect was observed in plants which were homozygous for antiGUS. RNA detection results suggest that duplex formation with the 41 base pair antiGUS RNA destabilized the GUS mRNA and that an excess of antisense. RNA was not required. Our results demonstrate the potential of antisense RNA as a strategy for obtaining plant mutants, especially ‘down mutations’ in essential genes where only a short 5′ sequence of the mRNA is required. They also suggest that the ‘position effect’ on gene expression could be used in conjunction with an antisense RNA strategy to provide a versatile approach for crop improvement.
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    Expression of a stilbene synthase gene in Nicotiana tabacum results in synthesis of the phytoalexin resveratrol (1990)
    Springer
    Plant molecular biology 15 (1990), S. 325-335 
    Details
    ISSN: 1573-5028
    Keywords: Nicotiana tabacum ; fungal disease resistance ; groundnut stilbene synthase ; gene expression ; resveratrol synthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A gene from groundnut (Arachis hypogaea) coding for stilbene synthase was transferred together with a chimaeric kanamycin resistance gene. It was found to be rapidly expressed after induction with UV light and elicitor in tobacco cells (Nicotiana tabacum). Comparative studies of stilbene synthase mRNA synthesis in groudnut and transgenic tobacco suspension cultures revealed the same kinetics of gene expression. Stilbene synthase specific mRNA was detectable 30 minutes after elicitor induction and 10 minutes after UV irradiation. The maximum of mRNA accumulation was between 2 and 8 hours post induction. 24 hours after induction stilbene synthase mRNA accumulation ceased. Furthermore, in transgenic tobacco plants, the gene was found to be inducible in sterile roots, stems and leaves. Stilbene synthase was demonstrated in crude protein extracts from transgenic tobacco cell cultures using specific antibodies. Resveratrol, the product of stilbene synthase, was identified by HPLC and antisera raised against resveratrol.
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    Tissue-specific expression of a cDNA clone from cell suspension cultures ofLupinus polyphyllus (1990)
    Springer
    Plant molecular biology 14 (1990), S. 1055-1056 
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    ISSN: 1573-5028
    Keywords: Lupinus polyphyllus ; cell culture ; cDNA clone ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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    URL: http://dx.doi.org/10.1007/BF00019403
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    A tomato cDNA inducible by salt stress and abscisic acid: nucleotide sequence and expression pattern (1990)
    Springer
    Plant molecular biology 15 (1990), S. 695-705 
    Details
    ISSN: 1573-5028
    Keywords: salt stress ; abscisic acid ; cDNA sequence ; gene expression ; LEA protein sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have characterized a new tomato cDNA, TAS14, inducible by salt stress and abscisic acid (ABA). Its nucleotide sequence predicts an open reading frame coding for a highly hydrophilic and glycine-rich (23.8%) protein of 130 amino acids. Southern blot analysis of tomato DNA suggests that there is one TAS14 structural gene per haploid genome. TAS14 mRNA accumulates in tomato seedlings upon treatment with NaCl, ABA or mannitol. It is also induced in roots, stems and leaves of hydroponically grown tomato plants treated with NaCl or ABA. TAS14 mRNA is not induced by other stress conditions such as cold and wounding. The sequence of the predicted TAS14 protein shows four structural domains similar to the rice RAB21, cotton LEA D11 and barley and maize dehydrin genes.
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    Genetic analysis of clathrin function in yeast (1990)
    Springer
    The journal of membrane biology 116 (1990), S. 93-105 
    Details
    ISSN: 1432-1424
    Keywords: clathrin ; genetics ; Saccharomyces cerevisiae ; exocytosis ; endocytosis ; prohormone maturation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
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    URL: http://dx.doi.org/10.1007/BF01868668
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    Genetic transfer, inheritance, and phenotypic expression of plasmid RP4::Mu cts genes inBacillus cereus (1990)
    Springer
    Bulletin of experimental biology and medicine 109 (1990), S. 389-392 
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    ISSN: 1573-8221
    Keywords: plasmid ; transception ; transcipient ; segregation ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
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    URL: http://dx.doi.org/10.1007/BF00839668
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    Molecular aspects of egg-laying behavior inAplysia californica (1990)
    Springer
    Behavior genetics 20 (1990), S. 251-264 
    Details
    ISSN: 1573-3297
    Keywords: egg-laying behavior ; Aplysia ; neuropeptides ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Psychology
    Notes: Abstract TheAplysia neuroendocrine system is a particularly advantageous model for cellular and molecular studies because of the relatively small number and large size of its component neurons. In addition, numerous anatomical and physiological studies have resulted in the assignment of behavioral roles to individual identified neurons. Recombinant DNA techniques have been used to isolate the genes that encode the precursors of peptides involved in egg-laying behavior. The comparison of the egg-laying hormone (ELH) gene family within the genusAplysia reveals high homologies in the overall structure of the precursors. A well-conserved tetrabasic residue has been shown to be the first endoproteolytic cleavage site of the precursor, giving rise to two intermediates, which are differentially processed and packaged. Some members of the ELH gene family are expressed specifically in the bag cell clusters or the atrial gland, respectively, providing an opportunity to study control of gene expression at the molecular level.
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    Ty RNA levels determine the spectrum of retrotransposition events that activate gene expression in Saccharomyces cerevisiae (1990)
    Springer
    Molecular genetics and genomics 220 (1990), S. 213-221 
    Details
    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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    Conjugative transfer and autonomous replication of a promiscuous IncQ plasmid in the cyanobacterium Synechocystis PCC 6803 (1990)
    Springer
    Molecular genetics and genomics 221 (1990), S. 129-133 
    Details
    ISSN: 1617-4623
    Keywords: Bacterial conjugation ; Cyanobacteria ; IncQ plasmids ; Saccharomyces cerevisiae ; DNA methylation
    Source: Springer Online Journal Archives 1860-2000
    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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    URL: http://dx.doi.org/10.1007/BF00280378
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    A GCN4 protein recognition element is not sufficient for GCN4-dependent regulation of transcription in the ARO7 promoter of Saccharomyces cerevisiae (1990)
    Springer
    Molecular genetics and genomics 224 (1990), S. 57-64 
    Details
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; AR07 gene ; Chorismate mutase ; GCN4 ; Transcriptional regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    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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    URL: http://dx.doi.org/10.1007/BF00259451
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    A yeast mutant, PRP20, altered in mRNA metabolism and maintenance of the nuclear structure, is defective in a gene homologous to the human gene RCC1 which is involved in the control of chromosome condensation (1990)
    Springer
    Molecular genetics and genomics 224 (1990), S. 72-80 
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    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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    The MSS51 gene product is required for the translation of the COX1 mRNA in yeast mitochondria (1990)
    Springer
    Molecular genetics and genomics 224 (1990), S. 111-118 
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    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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    Induction of “General Control” and thermotolerance in cdc mutants of Saccharomyces cerevisiae (1990)
    Springer
    Molecular genetics and genomics 224 (1990), S. 257-263 
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    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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    URL: http://dx.doi.org/10.1007/BF00271559
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    The UGA3 gene regulating the GABA catabolic pathway in Saccharomyces cerevisiae codes for a putative zinc-finger protein acting on RNA amount (1990)
    Springer
    Molecular genetics and genomics 220 (1990), S. 269-276 
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    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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    Regulation of chorismate mutase in Saccharomyces cerevisiae (1990)
    Springer
    Molecular genetics and genomics 220 (1990), S. 283-288 
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    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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    Differential regulation of STA genes of Saccharomyces cerevisiae (1990)
    Springer
    Molecular genetics and genomics 222 (1990), S. 87-96 
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    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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    Molecular basis for determining the sensitivity of eucaryotes to the antimitotic drug rhizoxin (1990)
    Springer
    Molecular genetics and genomics 222 (1990), S. 169-175 
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    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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    URL: http://dx.doi.org/10.1007/BF00633814
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    Correct splicing of modified introns of a Rhizopus proteinase gene in Saccharomyces cerevisiae (1990)
    Springer
    Molecular genetics and genomics 223 (1990), S. 11-16 
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    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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    Expression in Saccharomyces cerevisiae of a gene associated with cytoplasmic male sterility from maize: Respiratory dysfunction and uncoupling of yeast mitochondria (1990)
    Springer
    Molecular genetics and genomics 223 (1990), S. 24-32 
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    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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    Deletion analysis of domain independence in the TRP1 gene product of Neurospora crassa (1990)
    Springer
    Molecular genetics and genomics 223 (1990), S. 49-57 
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    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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    HSP12, a new small heat shock gene of Saccharomyces cerevisiae: Analysis of structure, regulation and function (1990)
    Springer
    Molecular genetics and genomics 223 (1990), S. 97-106 
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    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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    Interchromosomal and intrachromosomal recombination in rad 18 mutants of Saccharomyces cerevisiae (1990)
    Springer
    Molecular genetics and genomics 222 (1990), S. 25-32 
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    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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    DNA sequence elements required for transcription initiation of the Schizosaccharomyces pombe ADH gene in Saccharomyces cerevisiae (1990)
    Springer
    Molecular genetics and genomics 223 (1990), S. 407-416 
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    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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    TUF factor binds to the upstream region of the pyruvate decarboxylase structural gene (PDC1) of Saccharomyces cerevisiae (1990)
    Springer
    Molecular genetics and genomics 223 (1990), S. 449-456 
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    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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    Inducible cellular responses to ultraviolet light irradiation and other mediators of DNA damage in mammalian cells (1990)
    Springer
    Cell biology and toxicology 6 (1990), S. 105-126 
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    ISSN: 1573-6822
    Keywords: DNA damage ; DNA amplification ; ultraviolet light irradiation ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Both naturally occuring and carcinogen-induced tumors display not only point mutations in cellular oncogenes but also more complex changes in cellular oncogenes and other cellular genes. For this and other reasons, it seems likely that DNA damage in mammalian cells can induce alterations in gene expression that may have both short and long term consequences in the target cell. The purpose of this review is to summarize current available information on inducible responses to UV-irradiation and other mediators of DNA damage in mammalian cells, and to provide some working hypotheses. We have divided these responses into three time frames, immediate (0–12 hours), early (12–48) and late (beyond 48 hours). Immediate responses include the action of DNA repair enzymes, some of which are induced as a consequence of DNA damage, and transient inhibition of DNA synthesis. Within the past few years considerable evidence has accumulated that during this immediate period there is increased expression of certain cellular oncogenes, proteases and proteins whose functions remain to be identified. It is of interest that the expression of some of these genes is also induced by certain growth factors, tumor promoters and heat shock. Alterations in gene expression during the subsequent “early” period (12–48 hrs.) have not been studied in detail, but it is during this period that one can detect increased replication of several types of viruses in cells that harbor these viruses. We have examined in detail the induction of asynchronous polyoma DNA replication (APR) in a rat fibroblast cell line carrying integrated copies of this DNA. We have obtained evidence that UV-irradiation of these cells leads to the synthesis of a 40 kd protein, within the first 1–24 hrs after irradiation, that binds to a specific sequence TGACAACA in the regulatory region of polyoma DNA. We suggest that this protein acts together with other proteins to induce APR and that this serves as a useful model for understanding the mechanisms responsible for amplification of cellular genes, a phenomenon often seen in malignant tumors. Finally, we discuss how the events occurring during the immediate and early periods following DNA damage might lead to late effects in the target cell that are stable and contribute to the genotype and phenotype of some of the progeny of these cells that are destined to become tumor cells.
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    Neuropeptide gene expression and neural activity: Assessing a working hypothesis in nucleus caudalis and dorsal horn neurons expressing preproenkephalin and preprodynorphin (1990)
    Springer
    Cellular and molecular neurobiology 10 (1990), S. 73-98 
    Details
    ISSN: 1573-6830
    Keywords: molecular biology of peptidergic neurons ; in situ hybridization ; gene expression ; preproenkephalin ; preprodynorphin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 1. The working hypothesis that neuropeptide gene expression in a neuron is an indicator of that neuron's physiological activity is discussed. 2. Representative examples from the literature are presented to support the hypothesis. 3. Further, we discuss the regulation of expression of two opioid peptides, preproenkephalin and preprodynorphin, in laminae I and II of the spinal cord and in nucleus caudalis of the trigeminal nuclear complex, where they may play a role in pain modulation. 4. The expression of the opioid peptide genes can be induced by both painful and nonnoxious stimuli in neurons in time-dependent and sensory-specific fashions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00733637
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    Prenatal haloperidol alters the expression of DNA polymerases in brain regions of neonate rats (1990)
    Springer
    Cellular and molecular neurobiology 10 (1990), S. 281-289 
    Details
    ISSN: 1573-6830
    Keywords: haloperidol ; gene expression ; DNA polymerases ; brain development ; mesencephalon ; forebrain ; dopamine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 1. Previous studies have reported a marked reduction in the [3H]thymidine incorporation in forebrain after administration of a dopamine antagonist such as haloperidol. 2. We have investigated the possibility that the expression levels of genes related to DNA metabolism could be altered by haloperidol treatment. 3. By Northern blot analysis, we have studied the steady-state mRNA levels for genes involved in DNA metabolism, in neonate rat mesencephalon and forebrain, after chronic prenatal blockade of dopamine receptors with haloperidol. 4. We found that the expression levels for DNA polymerases alpha and beta were clearly reduced in forebrain by haloperidol treatment. On the contrary, the expression of DNA polymerase beta was increased in mesencephalon. 5. Our results suggest that dopamine receptors occupancy may be a critical factor in controlling cell proliferation during brain development, through a mechanism(s) involving changes in the expression of DNA polymerases.
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    URL: http://dx.doi.org/10.1007/BF00734581
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    Immunochemical analysis of the carbohydrate moiety of yeast killer toxin K28 (1990)
    Springer
    Antonie van Leeuwenhoek 58 (1990), S. 277-282 
    Details
    ISSN: 1572-9699
    Keywords: Saccharomyces cerevisiae ; killer toxin K28 ; glycoprotein ; O-glycosylation ; β-elimination ; α-toxin antibodies
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Killer toxin K28, a 16 kd protein secreted by the wine yeast Saccharomyces cerevisiae strain 28, was reversibly bound by a column of Concanavalin A-Sepharose, confirming its glycoprotein nature. HPLC analysis of acid hydrolyzates of K28 toxin as well as Western-blots of β-eliminated and/or endo H-treated killer toxin preparations probed with polyclonal α-toxin antibodies revealed that the carbohydrate moiety of K28 consists of D-mannose only, which is O-glycosidically linked via Ser/Thr residues to the protein part. The change in gel mobility of K28 after β-elimination was caused by a decrease in molecular mass of about 1,800, corresponding to a carbohydrate moiety of 10 mannose residues per killer toxin molecule.
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    URL: http://dx.doi.org/10.1007/BF00399340
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    Different regulation of mid-size neurofilament and N-myc mRNA expression during neuroblastoma cell differentiation induced by retinoic acid (1990)
    Springer
    Cellular and molecular neurobiology 10 (1990), S. 459-470 
    Details
    ISSN: 1573-6830
    Keywords: neuroblastoma ; differentiation ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 1. Neuroblastoma (NB) is an unusual neuroectodermal tumor showing a high degree of spontaneous regression. NB cells can be induced to differentiatein vitro by various agents. Cell differentiation results in morphological changes characteristic of the mature neuronal phenotype, including outgrowth of neuritelike structures with several interconnections. 2. Recent experiments indicate that morphological differentiation of NB cells is associated with changes in expression of N-myc, c-myc, and c-myb oncogenes and synthesis of neurofilament proteins. However, little is known about the transcription of neurofilament genes during differentiation. 3. We have analyzed the expression of both the N-myc oncogene and mid-size neurofilament (NF) genes in the LAN-1 human NB cell line, cultured in the presence of retinoic acid (RA). Continuous treatment with RA induced morphological differentiation within 5–6 days. The transcription of N-myc was down-modulated within 24 hr of the initial exposure to RA. The mid-size NF mRNA was increased at this time. The expression of N-myc was not modified in serum-deprived LAN-1 cells, indicating that N-myc transcription is unaffected by the arrest of the cells in the G1 phase. 4. We conclude that new synthesis of mid-size NF mRNA and a decrease in N-myc transcription precedede novo formation of neurite-like processes and morphological cell differentiation of neuroblastoma cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00711187
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    Cytoskeletal dynamics in rabbit synovial fibroblasts: I. Effects of acrylamide on intermediate filaments and microfilaments (1990)
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 16 (1990), S. 110-120 
    Details
    ISSN: 0886-1544
    Keywords: vimentin ; collagenase ; cell shape ; gene expression ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Rabbit synovial fibroblasts respond to changes in cell shape and cytoskeletal architecture by altering specific gene expression. We have tested the ability of acrylamide, a neurotoxin that alters the distribution of intermediate filaments in cultured PtKl cells, to induce metalloprotease expression in synovial fibroblasts. Cells treated with 2-20 mM acrylamide for 5 to 24 h underwent shape changes similar to cells treated with the tumor promoter phorbol myristate acetate. Intermediate filaments visualized with anti-vimentin antibodies did not collapse into a perinuclear cap in these rounded cells, but were still present in the extended cell processes. Unexpectedly, when actin was visualized in acrylamide-treated cells, extensive dissociation and clumping of microfilaments was observed. Concentrations of acrylamide 〉 10 mM were cytotoxic, but cells recovered completely after 24 h incubation with 5 mM acrylamide. Like other agents that alter cell shape and actin distribution in synovial fibroblasts, acrylamide also induced expression of the secreted metalloprotease collagenase. Although some recent evidence suggests that acrylamide may be able to exert its collagenase-inducing effects extra-cellularly, perhaps through transmembrane matrix receptors, our observation that this neurotoxin dramatically alters protein synthesis in synovial fibroblasts suggests that direct effects on cell metabolism may also play a role in acute acryl-amide intoxication.
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    URL: http://dx.doi.org/10.1002/cm.970160205
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    Toxicity of allelopathic monoterpene suspensions on yeast dependence on droplet size (1990)
    Springer
    Journal of chemical ecology 16 (1990), S. 1399-1408 
    Details
    ISSN: 1573-1561
    Keywords: Allelopathy ; emulsions ; monoterpenes ; Saccharomyces cerevisiae ; yeast ; suspensions ; droplet size ; toxicity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The toxic effects of the allelopathic nonsubstituted monoterpenes β-pinene and limonene on yeast,Saccharomyces cerevisiae, were proportional to the size of the monoterpene droplets in suspension. Both the toxic effects and the size of the droplets in suspension were decreased by adding different solvents with the monoterpene as follows: dimethylsulfoxide – dimethylformamide ≫ ethanol 〉 dioxane. Oxygen consumption was inhibited about 80% by 1 mM β-pinene added in dimethylsulfoxide but less than 10% when β-pinene was added in dioxane. Parallel decreases in droplet size and toxic effects of either monoterpene were also induced by hydrating the monoterpene-dimethylformamide or monoterpene-dimethylsulfoxide before addition to yeast. Molecular aggregation may be a mechanism to potentiate the allelopathic properties of monoterpenes when these associate with diverse soil components.
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    URL: http://dx.doi.org/10.1007/BF01021035
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    Expression and regulation of pOb24 and lipoprotein lipase genes during adipose conversion (1990)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 43 (1990), S. 103-110 
    Details
    ISSN: 0730-2312
    Keywords: gene expression ; differentiation ; adipose cell ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Lipoprotein lipase (LPL) and pOb24 mRNAs are known to be early markers of adipose cell differentiation. Comparative studies of the expression of pOb24 and LPL genes during adipose conversion of Ob1771 preadipocyte cells and in mouse adipose tissue have shown the following: (1) the expression of both genes takes place at confluence; this event can also be triggered by growth arrest of exponentially growing cells at the G1/S stage of the cell cycle; (2) In contrast to glycerol-3°phosphate dehydrogenase mRNA, the emergence of pOb24 and lipoprotein lipase mRNAs requires neither growth hormone or tri-iodothyronine as obligatory hormones nor insulin as a modulating hormone; (3) in mouse adipose tissue, pOb24 mRNA is present at a high level in stromal-vascular cells and at a low level in mature adipocytes, and in contrast LPL mRNAs are preferentially expressed in mature adipocytes. Thus, these two genes do not appear to be regulated in a similar manner, as also shown by the differential inhibition of their expression by tumor necrosis factor (TNF) and transforming growth factor-β (TGF-β ).
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    URL: http://dx.doi.org/10.1002/jcb.240430202
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    Protein synthesis in embryonic tissues during mouse postimplantation development (1990)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 44 (1990), S. 19-37 
    Details
    ISSN: 0730-2312
    Keywords: cell lineage ; tissue separation ; gene expression ; 2-D gels ; mammalian embryology ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Mouse embryos of the NMRI strain between the 7th and 9th day of gestation were isolated from the uterus and dissected into the various tissue derivatives in order to investigate newly synthesized proteins during morphogenesis. The day 7 embryo was fragmented into trophoblast and ectoplacental cone, distal and proximal endoderm, extraembryonic and embryonic ectoderm. The day 8 and day 9 embryos were divided into trophoblast and placental anlage, yolk sac, amnion, and allantois, as well as cranial, central, and caudal embryonic tissue. The intact embryos were incubated in Dulbecco's minimum essential medium in the presence of 35S-methionine for 4 h, then dissected into the various fragments, and further processed for two-dimensional gel electrophoresis. Protein synthesis of the isolated tissue derivatives was analyzed and compared for the three developmental stages. Concerning the proteins with isoelectric points in the range of 4.5 to 8.0 and molecular weight ratio (Mr) values between 20,000 and 200,000, we found several significant quantitative and qualitative differences in the various tissue fragments. In addition, we observed further quantitative and qualitative differences in protein synthesis during the postimplantation period investigated. We propose that the differences reflect some of the cell lineage- and developmental stage-specific changes in gene expression during early mammalian differentiation.
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    Fusion of luxA and luxB and its expression in E. coli, S. cerevisiae and D. melanogaster (1990)
    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 5 (1990), S. 89-97 
    Details
    ISSN: 0884-3996
    Keywords: Luciferase ; gene fusion ; Saccharomyces cerevisiae ; Drosophila melanogaster ; Vibrio harveyi ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Luciferase from Vibrio harveyi is encoded by two adjacent genes, luxA and luxB. The two genes were fused by replacing a segment extending from near the end of luxA into the N-terminal end of luxB by a synthetic oligonucleotide. The construction removed the TAA stop codon at the end of luxA, the intervening region of 26 base pairs, and the initial methionine of luxB. A Smal site was included at the junction between the two genes and an Aatll site was created near the end of luxA without altering its amino acid sequence. In Escherichia coli the fused luxAB gene could be expressed to produce functional luciferase that gave about 20% of the activity in cells without the fusion.An out-of frame ATG exists close to and preceding the ATG of the luxA gene. This was removed and the entire fused gene bracketed by several restriction enzyme sites.The fused luxAB gene was successfully expressed in Saccharomyces cerevisiae and Drosophila melanogaster by transferring it to appropriate plasmid vectors.
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    URL: http://dx.doi.org/10.1002/bio.1170050204
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    CCD imaging of luciferase gene expression in single mammalian cells (1990)
    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 5 (1990), S. 123-130 
    Details
    ISSN: 0884-3996
    Keywords: CCD ; imaging ; gene expression ; single cells ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Quantitative and sensitive imaging of chemiluminescence, bioluminescence and fluorescence emissions is emerging as an increasingly important technique for a range of biomedical applications (Hooper et al., 1990). A brief review of low-light-level imaging is presented, with particular reference to charge-coupled devices (CCD). Detectors for sensitive imaging are described and compared, including various CCDs and photoncounting devices. Image analysis techniques based on digital image processing, may be applied to quantify luminescent processes with these detectors. Images of luciferase gene expression in single mammalian cells have been obtained using a particular highsensitivity intensified CCD camera. The method is illustrated using cell monolayers infected with recombinant vaccinia virus encoding the firefly luciferase, luc gene (Rodriguez et al., 1988). The CCD camera has been used to detect luciferase expression in single, recombinant infected cells amongst over one million non-infected cells. The rapid detection of luciferase-expressing viruses may be used for the selection of virus deletion mutants into which the luciferase gene has been cloned at specific sites. This is particularly useful in the case of viruses such as cytomegalovirus which have slow replication cycles.This direct imaging technique is simple and versatile. It offers a rapid, non-invasive method for the sensitive detection of luciferase activity in single, luciferase-expressing cells. One can envisage the use of luciferase as a sensitive and convenient co-selection marker gene in the analysis of both gene expression and protein function. These methods offer tremendous potential in the fields of molecular and cellular biology.
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    URL: http://dx.doi.org/10.1002/bio.1170050208
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    Structural and functional analysis of the rat testis-specific histone H1t gene (1990)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 44 (1990), S. 1-17 
    Details
    ISSN: 0730-2312
    Keywords: histone genes ; gene structure ; gene expression ; histone mRNA ; rat liver ; rat testis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: A 6.86 kb rat genomic DNA fragment containing the testis-specific histone H1t gene and the histone H4t gene has been sequenced. S1-nuclease protection analyses of total cellular RNA from rat liver and testis showed that histone H1t mRNA was present only in testis. Examination of various highly enriched populations of rat testis cell types revealed that H1t mRNA was found exclusively in a fraction enriched in pachytene spermatocytes. When protein, DNA interactions within the proximal promoter region of the histone H1t gene were examined by electrophoretic mobility shift assays, only minor differences were found in mobility shift patterns of the H1t promoter in assays comparing binding of nuclear proteins from pachytene spermatocytes and early spermatids. However, major differences in binding were observed upon comparing nuclear proteins from rat pachytene spermatocytes to liver. Comparison of binding patterns of rat testis, rat hepatoma H4 cells, HeLa cells, and COS-1 cells also revealed dramatic differences. Transcriptional activity of the histone H1t promoter was examined by measuring H1t promoted chloramphenicol acetyltransferase (CAT) mRNA levels in transient experession assays in transfected rat hepatoma H4 cells, HeLa cells, and COS-1 cells. These assays revealed that the histone H1t promoted CAT gene functioned poorly in HeLa cells and COS-1 cells compared to expression with the parent SV40 promoted vector pSV2CAT. The H1t promoted CAT gene apparently did not work at all in transfected rat hepatoma H4 cells, which is consistent with testis germinal cell specific expression of the histone H1t gene.
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    Bacterial and firefly luciferase genes in transgenic plants: Advantages and disadvantages of a reporter gene (1990)
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 224-232 
    Details
    ISSN: 0192-253X
    Keywords: lux ; luc reporter genes ; light emission ; gene expression ; single photon imaging in vivo ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Genes encoding light-emitting luciferase were recently isolated from luminous marine bacteria and fireflies. Expression of luciferase genes in diverse organisms is a unique way for studying gene expression by simple and sensitive measurement of light. Recent advances in application of luciferase reporter genes are reviewed and documented by examples of in vivo visualization of their expression in transgenic plants.
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    URL: http://dx.doi.org/10.1002/dvg.1020110308
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    The start gene CDC28 and the genetic stability of yeast (1990)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 6 (1990), S. 231-243 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; cell cycle regulation ; CDC28 gene ; rho- mutation ; chromosome loss ; plasmid maintenance ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The cdc28-srm mutation in Saccheromyces cerevisiae decreases spontaneous and induced mitochondrial rhomutability and the mitotic stability of native chromosomes and recombinant circular minichromosomes. The effects of cdc28-srm on the genetic stability of cells support the hypothesis that links cell cycle regulation in yeast to changes in chromatin organization dependent on the start gene CDC28 (Hayles and Nurse, 1986).
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    Chromosome III of Saccharomyces cerevisiae: An ordered clone bank, a detailed restriction map and analysis of transcripts suggest the presence of 160 genes (1990)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 6 (1990), S. 383-401 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome III ; ordered clone bank ; physical map ; transcripts ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Using λ phage vector EMBL4, we isolated 344 clones containing segments of chromosome III of Saccharomyces cerevisiae, analysed their physical structure with eight restriction enzymes and sorted the data in contiguous groups with computer programmes. Furthermore, we performed Southern hybridizations between the sorted contiguous clone groups and interrelated them into larger groups. In this way, we constructed an ordered clone bank that covers almost the whole of chromosome III with a single gap of several kilobases in length. The consensus physical map thus obtained totals 334·6 kb, which is in good agreement with the size of this chromosome estimated by pulsed-field gel electrophoresis. Southern hybridization analysis with the DNA probes containing telomere-specific sequences showed that the bank contained a telomere at a position corresponding to the right arm terminus of chromosome III. Also, five Ty elements were found to be present. To estimate the number of genes on this chromosome and to analyse their levels of expression, we performed a series of Northern hybridization experiments using total poly(A)+ RNA from vegetatively growing cells and appropriate restriction enzyme fragments from the bank. Thus, we identified a total of 156 transcripts on chromosome III, indicating, on an average, one gene in every 2 kb on this chromosome. The transcripts were visually categorized into five groups according to their apparent levels of expression. It was found that the genes located near both termini are expressed only at low levels and that highly expressed genes are rather scattered over the chromosome.
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    Intracellular and extracellular levels of cyclic AMP during the cell cycle of Saccharomyces cerevisiae (1990)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 6 (1990), S. 53-60 
    Details
    ISSN: 0749-503X
    Keywords: Cyclic AMP ; Cell Cycle ; Saccharomyces cerevisiae ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Using the techniques of centrifugal elutriation it was demonstrated that during the cell division cycle of the budding yeast Saccharomyces cerevisiae there are stage-specific fluctuations in the intracellular concentration of adenosine 3′,5′-cyclic monophosphate (cAMP). Results shown here indicate that the intracellular concentration of cAMP is at its highest during the division cycle, and its lowest immediately prior to and just after cell sepraration. Results also show the extrusion of extracellular cAMP into the medium by Saccharomyces cerevisiae, extracellular cAMP levels being ten to one hundred times higher than intracellular levels. During the cell of Saccharomyces cerevisiae the extracellular level of cAMP does not fluctuate.
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    A novel aspartyl protease allowing KEX2-independent MFα propheromone processing in yeast (1990)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 6 (1990), S. 127-137 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; precursor processing ; aspartyl endopeptidase ; nucleotide sequence ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Mutants of Saccharomyces cerevisiae which lack the KEX2-encoded endopeptidase are unable to process proteolytically the mating factor alpha (MFα) propheromone produced from the chromosomal MFα1 and MFα2 genes (Julius et al., 1983). Overproduction of pheromone precursor from multiple, plasmid-borne MFα genes did, however, lead to the production of active MFα peptides in the absence of the KEX2 gene product. S. cerevisiae therefore must possess an alternative processing enzyme. The cleavage site of this enzyme appeared identical to that of the KEX2-encoded endopeptidase. To identify the gene responsible for the alternative processing, we have isolated clones which allowed production of mature MFα in a kex2-disrupted strain even from the chromosomal MFα genes. The gene isolated in this way was shown also to be essential for the KEX2-independent processing of propheromone overproduced from plasmid-borne MFα1. The amino acid sequence deduced from the gene shows extensive homology to a number of aspartyl proteases including the PEP4 and BARI gene products from S. cerevisiae. In contrast to the BARI gene product, the novel aspartyl protease (YAP3 for Yeast Aspartyl Protease 3) contains a C-terminal serine/threonine-rich sequence and potential transmembrane domain similar to those found in the KEX2 gene product. The corresponding gene YAP3 was located to chromosome XII. The normal physiological role of the YAP3 gene product is not known. Strains disrupted in YAP3 are both viable and able to process the mating factor a precursor.
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    Cell fusion of Saccharomyces cerevisiae fragile mutants (1990)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 6 (1990), S. 205-212 
    Details
    ISSN: 0749-503X
    Keywords: Cell fusion ; Saccharomyces cerevisiae ; fragile mutants ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Fragile mutants of Saccharomyces cerevisiae are defective in the structure of the cell wall and plasma membrane. The mutant cells lyse in hypotonic solutions but grow exponentially when osmotic stabilizer is induced in the medium. These mutants display a general increase in the permeability of the plasma membrane. We show here that fragile yeast cells of the same mating type can fuse without protoplast formation. The frequency of cell × cell fusion is lower than that observed for protoplast × protoplast fusion and can be significantly increased if the cells of one partner are converted to protoplasts. Microscopic observations and genetic analysis demonstrate that the hybrids obtained are fusion products. The fusion between fragile cells is explained in terms of the existence of local defects on their surface where the cell wall is thinner (or even missing), thus allowing a direct contact of cells by means of their plasma membranes.
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    Oscillatory CO2 evolution in glycolysing yeast extracts (1990)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 6 (1990), S. 255-261 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; glycolysis ; oscillation ; carbon dioxide ; mass spectrometry ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The rate of formation of carbon dioxide in cytoplasmic yeast extracts in an open system with continuous infusion of glucose was measured by membrane inlet mass spectrometry during glycolytic oscillations. The rate of CO2 production rose in the first third of each cycle to a maximum of about 100 μmol per ml yeast extract per hour and subsequently diminished to a final level of about 50 μmol per h. Measurements of the NADH light absorption under the same conditions revealed oscillations of relaxations type. The phase of high CO2 production could be related to the phase of the high NADH level, giving evidence that the flux in glycolysis is increased during the phase of high NADH concentration. Only half of the amount of injected glucose was metabolized to CO2 during the sustained oscillation, although free glucose did not accumulate.
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    Yeast Saccharomyces cerevisiae selectable markers in pUC18 polylinkers (1990)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 6 (1990), S. 363-366 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; Selectable markers ; Plasmids ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A set of plasmids was constructed that contain the yeast selectable markers HIS3, LEU2, TRP1 or URA3 embedded in the multiple cloning site of pUC18.
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    URL: http://dx.doi.org/10.1002/yea.320060502
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    The omnipotent suppressor SUP45 affects nucleic acid metabolism and mitochondrial structure (1990)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 6 (1990), S. 441-450 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; novobiocin-resistance ; SUP45 ; nucleic acid synthesis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Yeast (Saccharomyces cerevisiae) strains sensitive to a variety drugs were used to select for novobiocin-resistant mutants that were simultaneously temperature-sensitive. The mutants remained as sensitive as the parent strains to a wide range of drugs other than novobiocin, and did not exhibit any suppression of suppressible auxotrophic markers. At the non-permissive temperature, the mutant cells arrested mainly as unbudded cells, and were instantly defective in DNA and RNA synthesis, but not protein synthesis. The cloned wild-type gene was identified as SUP45, which has been previously implicated in the translation process. Our results suggest that SUP45 may have a function in addition to, or different from, the one that has been assigned to it previously.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320060509
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    Effect of leader primary structure on the translational efficiency of phosphoglycerate kinase mRNA in yeast (1990)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 6 (1990), S. 473-482 
    Details
    ISSN: 0749-503X
    Keywords: Leader ; phosphoglycerate kinase ; recombinant DNA ; Saccharomyces cerevisiae ; trailer ; translation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In order to determine the effect of nucleotide composition of the 5′-untranslated (leader) region on the translational efficiency of mRNA in yeast, we replaced a large part of the leader region of the phosphoglycerate kinase (PGK) gene by various deoxyoligonucleotides of defined sequence. All mutations left the context of the transcription initiations site and AUG start codon intact. The mutant genes were introduced into yeast cells on a multicopy vector and the ratio of the steady-state levels of PGKmRNA and protein were determined.We found the translational efficiency to be unaffected by the presence of either an 18 nucleotides (nt) long poly A or poly C tract or by sequences consisting of mixtures of A and C residues in any proportion. In contrast, a polyU tract, as well as mixtures of U and C residues, reduced translational efficiency by a factor of two to three, presumably by long-rang base -pairing between the leader and sequences elsewhere in the coding or 3′-non-coding regions of the messenger. In agreement with this hypothesis, a five-fold reduction in translational efficiency was found for an mRNA carrying a polyC tract in the leader as well as a polyG tract in the trailer, neither of which had any effect on translational efficiency by itself. Therefore, we conclude that the leader and trailer regions (including the polyA tail) of PGK mRNA are sufficiently close to base-pair when containing complementary sequences. The resulting secondary structure evidently constitutes a barrier for incoming 40S subunits on their way to the AUG start codon.The presence of an 18 nt long polyG tract in the leader completely abolished translation of the PGK mRNA in accordance with earlier observations. However, we found the leaders containing up to 40% G residues interspersed with either A or U, still allow highly efficient translation. This value is about four times as high as the average G content of leader sequence in naturally occurring yeast mRNAS.Finally, neither deletion of about 40% of the trailer sequence of PGK mRNA, not replacement of this sequence by homopolymer tracts had any effect on translational efficiency.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320060604
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    The chromosomal constitution of wine strains of Saccharomyces cerevisiae (1990)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 6 (1990), S. 367-382 
    Details
    ISSN: 0749-503X
    Keywords: Yeast genetics ; wine yeast ; Saccharomyces cerevisiae ; chromosomes ; karyotyping ; aneuploidy ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A general procedure is described for determining the chromosomal constitution of industrial strains of Saccharomyces cerevisiae based on analysis of segregation frequencies for input markers among random spore progeny of industrial-laboratory strain hybrids. The multiple auxotrophic haploid testers used carried a dominant erythromycin-resistance marker, allowing hybrids to be selected in mass matings with produced by the wild-type industrial strains. Analysis a number of independent crosses between the haploid testers and an unselected population of spores of each wine strain distinguished between disomic, trisomic and tetrasomic chromosomal complements in the parents. Possible explanations for a significant class of aberrant segregation frequencies are discussed.Results of the analysis indicate that UCD Enology 522 (Montrachet) is diployed and possibly trisomic for chromosome VII; 522X is diploid; UCD Enology 505 (California Champagne) is disomic for chromosome XVI, trisomic for chromosomes I, II, III, VI, VIII, IX, X, XII, XV, tetrasomic for chromosomes IV, XI, XIII, XIV and either trisomic or tetrasomic for chromosomes V and VII; and that UCD Enology 595 (Pasteur Champagne) is disomic for chromosomes I, II, III, IX, XVI, trisomic for chromosomes IV, VI, X, XII, XIV, XV, tetrasomic for chromosomes V, VIII, XI, XIII, and either disomic or tetrasomic for chromosome VII.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320060503
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    The yeast regulatory gene PHO4 encodes a helix-loop-helix motif (1990)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 6 (1990), S. 451-454 
    Details
    ISSN: 0749-503X
    Keywords: DNA-binding ; dimerization ; helix-loop-helix ; PHO4 activator ; Saccharomyces cerevisiae ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Evidence is presented, based on sequence comparisons and secondary structure prediction, of the presence of a DNA-binding and dimerization helix-loop-helix motif in the yeast transcription activator PHO4. Interest in the existence of this first known motif in yeast is discussed.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320060510
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    Expression of soybean lectin gene deletions in tobacco (1990)
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 160-167 
    Details
    ISSN: 0192-253X
    Keywords: Flanking sequences ; insertion events ; gene expression ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A series of constructs containing the developmentally regulated soybean lectin gene (Le1) were used to transform tobacco plants in order to assess developmental and quantitative regulation conferred by flanking sequences. The largest of the lectin constructs contained approximately 3,00 base pairs (bp) of Le1 5′ flanking region and 1,500 bp of the 3′ flanking region. The smallest construct contained no 5′ flanking region and 194 bp of the 3′ flanking region. ELISA assays of lectin in individual tobacco seeds and Southern blot analyses confirmed that most constructs were inherited as unique insertion events. Maximal expression of Le1 required more than 338 bp of 5′ sequence, indicating that far upstream factors are involved in quantitative control of lectin expression. Lectin expression declined more than 80% between deletions with 1,700 versus 338 bp of 5′ flanking sequence. In contrast, developmental control of lectin expression was maintained by Le1 inserts with only 190 bp of 5′ sequence. The lectin promoter offers a potential means to target high levels of gene expression to the developing seeds of soybean or other dicotyledonous plants.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/dvg.1020110206
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    Synthesis and transport of storage proteins by testes in Heliothis virescens (1990)
    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 14 (1990), S. 151-170 
    Details
    ISSN: 0739-4462
    Keywords: testis sheath ; gene expression ; protein transport ; spermatids ; mitochondria ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The synthesis of two storage protein subunits, 76,000-Mr and 82,000-Mr polypeptides, by the testes sheath has been studied in Heliothis virescens. Like fat body, which is the primary site of synthesis for the large extratesticular pool, cells of the testes sheath secrete glycosylated storage proteins assembled into hexamers. The testis sheath differed from fat body in several important respects, including the failure to synthesize an abundant (in the hemolymph) 74,000-Mr storage protein, its relatively reduced expression of the 76,000-Mr polypeptide, and the absence of resorption of storage proteins from the lumen of the testis during pupal development. Cyst cells were also shown to import actively the 82,000-Mr storage protein by pinocytosis of testicular fluid and transfer it to the developing spermatids. Unlike other cell types that sequester storage proteins in the form of cytoplasmic granules, their localization within spermatids was exclusively mitochondrial. These observations suggest that expression of the storage protein genes is regulated tissue specifically and reveal novel pathways for their transport and, perhaps, utilization and function during development.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/arch.940140304
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