Isolation, DNA sequence, and regulation of a Saccharomyces cerevisiae gene that encodes DNA strand transfer protein alpha

DNA strand transfer protein alpha (STP alpha) from meiotic Saccharomyces cerevisiae cells promotes homologous pairing of DNA without any nucleotide cofactor in the presence of yeast single-stranded DNA binding protein. This gene (DNA strand transferase 1, DST1) encodes a 309-amino-acid protein with a predicted molecular mass of 34,800 Da. The STP alpha protein level is constant in both mitotic and meiotic cells, but during meiosis the polypeptide is activated by an unknown mechanism, resulting in a large increase in its specific activity. A dst1::URA3/dst1::URA3 mutant grows normally in mitotic media; however, meiotic cells exhibit a greatly reduced induction of both DNA strand transfer activity and intragenic recombination between his1 heteroalleles. Spore viability is normal. These results suggest that DST1 is required for much of the observed induction of homologous recombination in S. cerevisiae during meiosis but not for normal sporulation.

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Cloning and characterization of DST2, the gene for DNA strand transfer protein beta from Saccharomyces cerevisiae.

Molecular and Cellular Biology ◽

10.1128/mcb.11.5.2583 ◽

1991 ◽

Vol 11 (5) ◽

pp. 2583-2592 ◽

Cited By ~ 57

Author(s):

C C Dykstra ◽

K Kitada ◽

A B Clark ◽

R K Hamatake ◽

A Sugino

Keyword(s):

Saccharomyces Cerevisiae ◽

Intragenic Recombination ◽

Temperature Sensitive ◽

Strand Transfer ◽

Gene Encoding ◽

Yeast Saccharomyces Cerevisiae ◽

Prolonged Incubation ◽

Transfer Protein ◽

Dna Strand

The gene encoding the 180-kDa DNA strand transfer protein beta from the yeast Saccharomyces cerevisiae was identified and sequenced. This gene, DST2 (DNA strand transferase 2), was located on chromosome VII. dst2 gene disruption mutants exhibited temperature-sensitive sporulation and a 50% longer generation time during vegetative growth than did the wild type. Spontaneous mitotic recombination in the mutants was reduced severalfold for both intrachromosomal recombination and intragenic gene conversion. The mutants also had reduced levels of the intragenic recombination that is induced during meiosis. Meiotic recombinants were, however, somewhat unstable in the mutants, with a decrease in recombinants and survival upon prolonged incubation in sporulation media. spo13 or spo13 rad50 mutations did not relieve the sporulation defect of dst2 mutations. A dst1 dst2 double mutant has the same phenotype as a dst2 single mutant. All phenotypes associated with the dst2 mutations could be complemented by a plasmid containing DST2.

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Cloning and characterization of DST2, the gene for DNA strand transfer protein beta from Saccharomyces cerevisiae

Molecular and Cellular Biology ◽

10.1128/mcb.11.5.2583-2592.1991 ◽

1991 ◽

Vol 11 (5) ◽

pp. 2583-2592 ◽

Cited By ~ 1

Author(s):

C C Dykstra ◽

K Kitada ◽

A B Clark ◽

R K Hamatake ◽

A Sugino

Keyword(s):

Saccharomyces Cerevisiae ◽

Intragenic Recombination ◽

Temperature Sensitive ◽

Strand Transfer ◽

Gene Encoding ◽

Yeast Saccharomyces Cerevisiae ◽

Prolonged Incubation ◽

Transfer Protein ◽

Dna Strand

The gene encoding the 180-kDa DNA strand transfer protein beta from the yeast Saccharomyces cerevisiae was identified and sequenced. This gene, DST2 (DNA strand transferase 2), was located on chromosome VII. dst2 gene disruption mutants exhibited temperature-sensitive sporulation and a 50% longer generation time during vegetative growth than did the wild type. Spontaneous mitotic recombination in the mutants was reduced severalfold for both intrachromosomal recombination and intragenic gene conversion. The mutants also had reduced levels of the intragenic recombination that is induced during meiosis. Meiotic recombinants were, however, somewhat unstable in the mutants, with a decrease in recombinants and survival upon prolonged incubation in sporulation media. spo13 or spo13 rad50 mutations did not relieve the sporulation defect of dst2 mutations. A dst1 dst2 double mutant has the same phenotype as a dst2 single mutant. All phenotypes associated with the dst2 mutations could be complemented by a plasmid containing DST2.

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Genetic evidence for preferential strand transfer during meiotic recombination in yeast.

Genetics ◽

10.1093/genetics/125.4.753 ◽

1990 ◽

Vol 125 (4) ◽

pp. 753-761 ◽

Cited By ~ 1

Author(s):

D K Nag ◽

T D Petes

Keyword(s):

Saccharomyces Cerevisiae ◽

Meiotic Recombination ◽

Exchange Process ◽

Genetic Evidence ◽

The Other ◽

Strand Transfer ◽

Yeast Saccharomyces Cerevisiae ◽

Dna Strand

Abstract During meiotic recombination in the yeast Saccharomyces cerevisiae, heteroduplexes are formed as an intermediate in the exchange process. In the formation of an asymmetric heteroduplex, one chromosome acts as a donor of a single DNA strand and the other acts as a recipient. We present genetic evidence that the nontranscribed strand is donated more frequently than the transcribed strand in spores that have an unrepaired mismatch at the HIS4 locus.

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Molecular mechanism of autophagy in yeast, Saccharomyces cerevisiae

Philosophical Transactions of the Royal Society B Biological Sciences ◽

10.1098/rstb.1999.0501 ◽

1999 ◽

Vol 354 (1389) ◽

pp. 1577-1581 ◽

Cited By ~ 35

Author(s):

Y. Ohsumi

Keyword(s):

Saccharomyces Cerevisiae ◽

Amino Acid ◽

Conjugation System ◽

Yeast Saccharomyces Cerevisiae ◽

Whole Process ◽

Unique Protein ◽

Acid Protein ◽

Major Route ◽

Lytic Compartment ◽

Amino Acid Protein

Bulk degradation of cytosol and organelles is important for cellular homeostasis under nutrient limitation, cell differentiation and development. This process occurs in a lytic compartment, and autophagy is the major route to the lysosome and/or vacuole. We found that yeast, Saccharomyces cerevisiae , induces autophagy under various starvation conditions. The whole process is essentially the same as macroautophagy in higher eukaryotic cells. However, little is known about the mechanism of autophagy at a molecular level. To elucidate the molecules involved, a genetic approach was carried out and a total of 16 autophagy–defective mutants ( apg ) were isolated. So far, 14 APG genes have been cloned. Among them we recently found a unique protein conjugation system essential for autophagy. The C–terminal glycine residue of a novel modifier protein Apg12p, a 186–amino–acid protein, is conjugated to a lysine residue of Apg5p, a 294–amino–acid protein, via an isopeptide bond. We also found that apg7 and apg10 mutants were unable to form an Apg12p–Apg5p conjugate. The conjugation reaction is mediated via Apg7p, E1–like activating enzyme and Apg10p, indicating that it is a ubiquitination–like system. These APG genes have mammalian homologues, suggesting that the Apg12 system is conserved from yeast to human. Further molecular and cell biological analyses of APG gene products will give us crucial clues to uncover the mechanism and regulation of autophagy.

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Suppression of mutations in two Saccharomyces cerevisiae genes by the adenovirus E1A protein.

Molecular and Cellular Biology ◽

10.1128/mcb.15.6.3227 ◽

1995 ◽

Vol 15 (6) ◽

pp. 3227-3237 ◽

Cited By ~ 22

Author(s):

H A Zieler ◽

M Walberg ◽

P Berg

Keyword(s):

Saccharomyces Cerevisiae ◽

Amino Acid ◽

Single Gene ◽

Adenovirus E1a ◽

C Terminus ◽

Acid Protein ◽

Conserved Region ◽

Mutant Phenotypes ◽

Yeast Genes ◽

Amino Acid Protein

The protein products of the adenoviral E1A gene are implicated in a variety of transcriptional and cell cycle events, involving interactions with several proteins present in human cells, including parts of the transcriptional machinery and negative regulators of cell division such as the Rb gene product and p107. To determine if there are functional homologs of E1A in Saccharomyces cerevisiae, we have developed a genetic screen for mutants that depend on E1A for growth. The screen is based on a colony color sectoring assay which allows the identification of mutants dependent on the maintenance and expression of an E1A-containing plasmid. Using this screen, we have isolated five mutants that depend on expression of the 12S or 13S cDNA of E1A for growth. A plasmid shuffle assay confirms that the plasmid-dependent phenotype is due to the presence of either the 12S or the 13S E1A cDNA and that both forms of E1A rescue growth of all mutants equally well. The five mutants fall into two classes that were named web1 and web2 (for "wants E1A badly"). Plasmid shuffle assays with mutant forms of E1A show that conserved region 1 (CR1) is required for rescue of the growth of the web1 and web2 E1A-dependent yeast mutants, while the N-terminal 22 amino acids are only partially required; conserved region 2 (CR2) and the C terminus are dispensable. The phenotypes of mutants in both the web1 and the web2 groups are due to a single gene defect, and the yeast genes that fully complement the mutant phenotypes of both groups were cloned. The WEB1 gene sequence encodes a 1,273-amino-acid protein that is identical to SEC31, a protein involved in the budding of transport vesicles from the endoplasmic reticulum. The WEB2 gene encodes a 1,522-amino-acid protein with homology to nucleic acid-dependent ATPases. Deletion of either WEB1 or WEB2 is lethal. Expression of E1A is not able to rescue the lethality of either the web1 or the web2 null allele, implying allele-specific mutations that lead to E1A dependence.

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Repression of the genes for lysine biosynthesis in Saccharomyces cerevisiae is caused by limitation of Lys14-dependent transcriptional activation

Molecular and Cellular Biology ◽

10.1128/mcb.14.10.6411-6418.1994 ◽

1994 ◽

Vol 14 (10) ◽

pp. 6411-6418

Author(s):

A Feller ◽

E Dubois ◽

F Ramos ◽

A Piérard

Keyword(s):

Saccharomyces Cerevisiae ◽

Dna Binding ◽

Transcriptional Activation ◽

Activation Function ◽

Lysine Biosynthesis ◽

Binding Motif ◽

Terminal Portion ◽

Rich Domain ◽

Acid Protein ◽

Amino Acid Protein

The product of the LYS14 gene of Saccharomyces cerevisiae activates the transcription of at least four genes involved in lysine biosynthesis. Physiological and genetic studies indicate that this activation is dependent on the inducer alpha-aminoadipate semialdehyde, an intermediate of the pathway. The gene LYS14 was sequenced and, from its nucleotide sequence, predicted to encode a 790-amino-acid protein carrying a cysteine-rich DNA-binding motif of the Zn(II)2Cys6 type in its N-terminal portion. Deletion of this N-terminal portion including the cysteine-rich domain resulted in the loss of LYS14 function. To test the function of Lys14 as a transcriptional activator, this protein without its DNA-binding motif was fused to the DNA-binding domain of the Escherichia coli LexA protein. The resulting LexA-Lys14 hybrid protein was capable of activating transcription from a promoter containing a lexA operator, thus confirming the transcriptional activation function of Lys14. Furthermore, evidence that this function, which is dependent on the presence of alpha-aminoadipate semialdehyde, is antagonized by lysine was obtained. Such findings suggest that activation by alpha-aminoadipate semialdehyde and the apparent repression by lysine are related mechanisms. Lysine possibly acts by limiting the supply of the coinducer, alpha-aminoadipate semialdehyde.

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Sequence and nuclear localization of the Saccharomyces cerevisiae HAP2 protein, a transcriptional activator

Molecular and Cellular Biology ◽

10.1128/mcb.7.2.578-585.1987 ◽

1987 ◽

Vol 7 (2) ◽

pp. 578-585

Author(s):

J L Pinkham ◽

J T Olesen ◽

L P Guarente

Keyword(s):

Saccharomyces Cerevisiae ◽

Dna Sequence ◽

Fusion Gene ◽

Protein A ◽

Acid Protein ◽

Genes Encoding ◽

Beta Galactosidase ◽

Amino Acid Protein ◽

Operator Site ◽

Direct Activator

Activation of the CYC1 upstream activation site (UAS2) and other Saccharomyces cerevisiae genes encoding respiratory functions requires the products of the regulatory loci HAP2 and HAP3. We present here the DNA sequence of the yeast HAP2 gene and an initial investigation into the function of its product. The DNA sequence indicated that HAP2 encoded a 265-amino-acid protein whose carboxyl third was highly basic. Also found in the sequence was a polyglutamine tract spanning residues 120 to 133. Several experiments described herein suggest that HAP2 encodes a direct activator of transcription. First, a bifunctional HAP2-beta-galactosidase fusion gene was localized to the yeast nucleus. Second, a lexA-HAP2 fusion gene was capable of activating transcription when bound to a lexA operator site. The additional requirement for the HAP3 product in activation is discussed.

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The DAL81 gene product is required for induced expression of two differently regulated nitrogen catabolic genes in Saccharomyces cerevisiae

Molecular and Cellular Biology ◽

10.1128/mcb.11.2.1161-1166.1991 ◽

1991 ◽

Vol 11 (2) ◽

pp. 1161-1166 ◽

Cited By ~ 1

Author(s):

P A Bricmont ◽

J R Daugherty ◽

T G Cooper

Keyword(s):

Saccharomyces Cerevisiae ◽

Steady State ◽

Amino Acid ◽

The Other ◽

Loss Of Function ◽

Induced Expression ◽

Catabolic Genes ◽

Acid Protein ◽

Steady State Levels ◽

Amino Acid Protein

We demonstrate that the DAL81 gene, previously thought to be specifically required for induced expression of the allantoin pathway genes in Saccharomyces cerevisiae, functions in a more global manner. The data presented show it to be required for utilization of 4-aminobutyrate as a nitrogen source and for 4-aminobutyrate-induced increases in the steady-state levels of UGA1 mRNA. The DAL81 gene encodes a 970-amino-acid protein containing sequences homologous to the Zn(II)2Cys6 motif and two stretches of polyglutamine residues. Deletion of sequences homologous to the Zn(II)2Cys6 motif did not result in a detectable loss of function. On the other hand, loss of one of the polyglutamine stretches, but not the other, resulted in a 50% loss of DAL81 function.

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TEC1, a gene involved in the activation of Ty1 and Ty1-mediated gene expression in Saccharomyces cerevisiae: cloning and molecular analysis.

Molecular and Cellular Biology ◽

10.1128/mcb.10.7.3541 ◽

1990 ◽

Vol 10 (7) ◽

pp. 3541-3550 ◽

Cited By ~ 68

Author(s):

I Laloux ◽

E Dubois ◽

M Dewerchin ◽

E Jacobs

Keyword(s):

Gene Expression ◽

Saccharomyces Cerevisiae ◽

Amino Acid ◽

Molecular Analysis ◽

Mating Type ◽

Gene Activation ◽

Adjacent Gene ◽

Transcript Levels ◽

Acid Protein ◽

Amino Acid Protein

Ty and Ty-mediated gene expression observed in haploid cells of Saccharomyces cerevisiae depends on several determinants, some of which are required for the expression of haploid-specific genes. We report here the cloning and molecular analysis of TEC1. TEC1 encodes a 486-amino-acid protein that is a trans-acting factor required for full Ty1 expression and Ty1-mediated gene activation. However, mutation or deletion of the TEC1 gene had little effect on total Ty2 transcript levels. Our analysis provides clear evidence that TEC1 is not involved in mating or sporulation processes. Unlike most of the proteins involved in Ty and adjacent gene expression, the product of TEC1 has no known cellular function. Although there was no mating-type effect on TEC1 expression, our results indicate that the TEC1 and the a/alpha diploid controls on Ty1 expression are probably not cumulative.

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