scholarly journals Molecular cloning and characterization of the STE7 and STE11 genes of Saccharomyces cerevisiae.

1985 ◽  
Vol 5 (8) ◽  
pp. 1878-1886 ◽  
Author(s):  
D T Chaleff ◽  
K Tatchell
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Molecular Cloning ◽  
Mating Type ◽  
Cell Types ◽  
Transcriptional Level ◽  
Cell Type ◽  
Yeast Saccharomyces Cerevisiae ◽  
Genetic Techniques ◽  
Conjugation Process ◽  
Mat Locus

In the yeast Saccharomyces cerevisiae, haploid cells occur in one of the two cell types, a or alpha. The allele present at the mating type (MAT) locus plays a prominent role in the control of cell type expression. An important consequence of the elaboration of cell type is the ability of cells of one mating type to conjugate with cells of the opposite mating type, resulting in yet a third cell type, an a/alpha diploid. Numerous genes that are involved in the expression of cell type and the conjugation process have been identified by standard genetic techniques. Molecular analysis has shown that expression of several of these genes is subject to control on the transcriptional level by the MAT locus. Two genes, STE7 and STE11, are required for mating in both haploid cell types; ste7 and ste11 mutants are sterile. We report here the molecular cloning of STE7 and STE11 genes and show that expression of these genes is not regulated transcriptionally by the MAT locus. We also have genetically mapped the STE11 gene to chromosome XII, 40 centimorgans from ura4.

Molecular cloning and characterization of the STE7 and STE11 genes of Saccharomyces cerevisiae

1985 ◽  
Vol 5 (8) ◽  
pp. 1878-1886
Author(s):  
D T Chaleff ◽  
K Tatchell
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Molecular Cloning ◽  
Mating Type ◽  
Cell Types ◽  
Transcriptional Level ◽  
Cell Type ◽  
Yeast Saccharomyces Cerevisiae ◽  
Genetic Techniques ◽  
Conjugation Process ◽  
Mat Locus

In the yeast Saccharomyces cerevisiae, haploid cells occur in one of the two cell types, a or alpha. The allele present at the mating type (MAT) locus plays a prominent role in the control of cell type expression. An important consequence of the elaboration of cell type is the ability of cells of one mating type to conjugate with cells of the opposite mating type, resulting in yet a third cell type, an a/alpha diploid. Numerous genes that are involved in the expression of cell type and the conjugation process have been identified by standard genetic techniques. Molecular analysis has shown that expression of several of these genes is subject to control on the transcriptional level by the MAT locus. Two genes, STE7 and STE11, are required for mating in both haploid cell types; ste7 and ste11 mutants are sterile. We report here the molecular cloning of STE7 and STE11 genes and show that expression of these genes is not regulated transcriptionally by the MAT locus. We also have genetically mapped the STE11 gene to chromosome XII, 40 centimorgans from ura4.

scholarly journals Pheromones and pheromone receptors are the primary determinants of mating specificity in the yeast Saccharomyces cerevisiae.

Genetics ◽  
1989 ◽  
Vol 121 (3) ◽  
pp. 463-476 ◽  
Author(s):  
A Bender ◽  
G F Sprague
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Types ◽  
Haploid Cell ◽  
Wild Type ◽  
Cell Type ◽  
Yeast Saccharomyces Cerevisiae ◽  
A Cells ◽  
Alpha Factor ◽  
A Cell ◽  
Specific Proteins

Abstract Saccharomyces cerevisiae has two haploid cell types, a and alpha, each of which produces a unique set of proteins that participate in the mating process. We sought to determine the minimum set of proteins that must be expressed to allow mating and to confer specificity. We show that the capacity to synthesize alpha-factor pheromone and a-factor receptor is sufficient to allow mating by mat alpha 1 mutants, mutants that normally do not express any alpha- or a-specific products. Likewise, the capacity to synthesize a-factor receptor and alpha-factor pheromone is sufficient to allow a ste2 ste6 mutants, which do not produce the normal a cell pheromone and receptor, to mate with wild-type a cells. Thus, the a-factor receptor and alpha-factor pheromone constitute the minimum set of alpha-specific proteins that must be produced to allow mating as an alpha cell. Further evidence that the pheromones and pheromone receptors are important determinants of mating specificity comes from studies with mat alpha 2 mutants, cells that simultaneously express both pheromones and both receptors. We created a series of strains that express different combinations of pheromones and receptors in a mat alpha 2 background. These constructions reveal that mat alpha 2 mutants can be made to mate as either a cells or as alpha cells by causing them to express only the pheromone and receptor set appropriate for a particular cell type. Moreover, these studies show that the inability of mat alpha 2 mutants to respond to either pheromone is a consequence of two phenomena: adaptation to an autocrine response to the pheromones they secrete and interference with response to alpha factor by the a-factor receptor.

scholarly journals INTERCONVERSION OF YEAST CELL TYPES BY TRANSPOSABLE GENES

Genetics ◽  
1980 ◽  
Vol 95 (3) ◽  
pp. 631-648
Author(s):  
Amar J S Klar
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Yeast Cell ◽  
Mating Type ◽  
Genetic Information ◽  
Cell Types ◽  
Yeast Saccharomyces Cerevisiae ◽  
Type Locus ◽  
Mating Type Switching ◽  
Controlling Element ◽  
Unidirectional Transfer

ABSTRACT The a and α cell types of budding yeast Saccharomyces cerevisiae are controlled by alternate alleles of the mating-type locus (MAT), MATa and MATα. The cell types can be interconverted by switching alleles of MAT. The loci HMRa and HMLα, which are loosely linked to MAT, are involved in mating-type switching. Experimental evidence for their role in MAT interconversion is presented. As a result of switching, the homothallic and heterothallic strains containing the amber and ochre mutations within the HMRa locus yield corresponding amber and ochre mutant mata loci. Similarly, the hmlα mutant strain generates mata mutant alleles. That is, specific mutations from HMRa and HMLα are transmitted to MAT. A replica of the mating-type coding information originating from these loci is transposed to MAT, where it replaces the existing information. Furthermore, "Hawthorne deletions" in strains containing hmra-ambe/ochre result in production of mata-ambedochre alleles, Therefore, genetic information for MATa resides at HMRa. The switches occur in a defined set of clonally related cells. Thus, the efficient interconversion of yeast cell types is mediated by an unidirectional transfer of genetic information between nonallelic sites in a nonrandom and programmed fashion. The results are inconsistent with the "flipflop" models, but satisfy a key prediction of the general controlling element and the specific cassette models proposed for mating-type interchange.

scholarly journals IME4, a gene that mediates MAT and nutritional control of meiosis in Saccharomyces cerevisiae.

1992 ◽  
Vol 12 (3) ◽  
pp. 1078-1086 ◽  
Author(s):  
J C Shah ◽  
M J Clancy
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Nutrient Availability ◽  
Cell Types ◽  
Cell Type ◽  
Transcript Accumulation ◽  
Yeast Saccharomyces Cerevisiae ◽  
New Gene ◽  
Low Levels ◽  
Diploid Cells

In the yeast Saccharomyces cerevisiae, sporulation occurs in response to nutritional and genetic signals. The process is initiated when nutrient availability limits mitotic growth, but only in MATa/MAT alpha diploid cells. Under these conditions, the cells express an activator of meiosis (IME1), which is required for the expression of early sporulation-specific genes. We describe a new gene, IME4, whose activity is essential for IME1 transcript accumulation and sporulation. The IME4 transcript was induced in starved MATa/MAT alpha diploids but not in other cell types. In addition, excess IME4 promoted sporulation in mat-insufficient cells. Thus, IME4 appears to activate IME1 in response to cell type and nutritional signals. We have also explored the interactions between IME4 and two genes that are known to regulate IME1 expression. Normally, cells that lack complete MAT information cannot sporulate; when such strains lack RME1 activity or contain the semidominant RES1-1 mutation, however, they can express IME1 and sporulate to low levels. Our results show that mat-insufficient strains containing rme1::LEU2 or RES1-1 bypass mutations still retain MAT control of IME4 expression. Even though IME4 levels remained low, the rme1::LEU2 and RES1-1 mutations allowed IME1 accumulation, implying that these mutations do not require IME4 to exert their effects. In accord with this interpretation, the RES1-1 mutation allowed IME1 accumulation in MATa/MAT alpha strains that contain ime4::LEU2 alleles. These strains still sporulated poorly, suggesting that IME4 plays a role in sporulation in addition to promoting IME1 transcript accumulation. IME4 is located between ADE5 and LYS5 on chromosome VII.

scholarly journals Donor locus selection during Saccharomyces cerevisiae mating type interconversion responds to distant regulatory signals.

Genetics ◽  
1992 ◽  
Vol 132 (4) ◽  
pp. 929-942
Author(s):  
K S Weiler ◽  
J R Broach
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mating Type ◽  
Positional Information ◽  
Yeast Saccharomyces Cerevisiae ◽  
Preferential Selection ◽  
Locus Selection ◽  
Chromosome Iii ◽  
Mat Locus ◽  
Donor Locus ◽  
Selection Of

Abstract Mating type interconversion in homothallic strains of the yeast Saccharomyces cerevisiae results from directed transposition of a mating type allele from one of the two silent donor loci, HML and HMR, to the expressing locus, MAT. Cell type regulates the selection of the particular donor locus to be utilized during mating type interconversion: MATa cells preferentially select HML alpha and MAT alpha cells preferentially select HMRa. Such preferential selection indicates that the cell is able to distinguish between HML and HMR during mating type interconversion. Accordingly, we designed experiments to identify those features perceived by the cell to discriminate HML and HMR. We demonstrate that discrimination does not derive from the different structures of the HML and HMR loci, from the unique sequences flanking each donor locus nor from any of the DNA distal to the HM loci on chromosome III. Moreover, we find that the sequences flanking the MAT locus do not function in the preferential selection of one donor locus over the other. We propose that the positions of the donor loci on the left and right arms of chromosome III is the characteristic utilized by the cell to distinguish HML and HMR. This positional information is not generated by either CEN3 or the MAT locus, but probably derives from differences in the chromatin structure, chromosome folding or intranuclear localization of the two ends of chromosome III.

IME4, a gene that mediates MAT and nutritional control of meiosis in Saccharomyces cerevisiae

1992 ◽  
Vol 12 (3) ◽  
pp. 1078-1086
Author(s):  
J C Shah ◽  
M J Clancy
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Nutrient Availability ◽  
Cell Types ◽  
Cell Type ◽  
Transcript Accumulation ◽  
Yeast Saccharomyces Cerevisiae ◽  
New Gene ◽  
Low Levels ◽  
Diploid Cells

In the yeast Saccharomyces cerevisiae, sporulation occurs in response to nutritional and genetic signals. The process is initiated when nutrient availability limits mitotic growth, but only in MATa/MAT alpha diploid cells. Under these conditions, the cells express an activator of meiosis (IME1), which is required for the expression of early sporulation-specific genes. We describe a new gene, IME4, whose activity is essential for IME1 transcript accumulation and sporulation. The IME4 transcript was induced in starved MATa/MAT alpha diploids but not in other cell types. In addition, excess IME4 promoted sporulation in mat-insufficient cells. Thus, IME4 appears to activate IME1 in response to cell type and nutritional signals. We have also explored the interactions between IME4 and two genes that are known to regulate IME1 expression. Normally, cells that lack complete MAT information cannot sporulate; when such strains lack RME1 activity or contain the semidominant RES1-1 mutation, however, they can express IME1 and sporulate to low levels. Our results show that mat-insufficient strains containing rme1::LEU2 or RES1-1 bypass mutations still retain MAT control of IME4 expression. Even though IME4 levels remained low, the rme1::LEU2 and RES1-1 mutations allowed IME1 accumulation, implying that these mutations do not require IME4 to exert their effects. In accord with this interpretation, the RES1-1 mutation allowed IME1 accumulation in MATa/MAT alpha strains that contain ime4::LEU2 alleles. These strains still sporulated poorly, suggesting that IME4 plays a role in sporulation in addition to promoting IME1 transcript accumulation. IME4 is located between ADE5 and LYS5 on chromosome VII.

scholarly journals Cloning and characterization of four SIR genes of Saccharomyces cerevisiae.

1986 ◽  
Vol 6 (2) ◽  
pp. 688-702 ◽  
Author(s):  
J M Ivy ◽  
A J Klar ◽  
J B Hicks
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mating Type ◽  
Blot Analysis ◽  
Transcriptional Control ◽  
Genomic Library ◽  
Yeast Saccharomyces Cerevisiae ◽  
Mating Type Genes ◽  
Rna Blots

Mating type in the yeast Saccharomyces cerevisiae is determined by the MAT (a or alpha) locus. HML and HMR, which usually contain copies of alpha and a mating type information, respectively, serve as donors in mating type interconversion and are under negative transcriptional control. Four trans-acting SIR (silent information regulator) loci are required for repression of transcription. A defect in any SIR gene results in expression of both HML and HMR. The four SIR genes were isolated from a genomic library by complementation of sir mutations in vivo. DNA blot analysis suggests that the four SIR genes share no sequence homology. RNA blots indicate that SIR2, SIR3, and SIR4 each encode one transcript and that SIR1 encodes two transcripts. Null mutations, made by replacement of the normal genomic allele with deletion-insertion mutations created in the cloned SIR genes, have a Sir- phenotype and are viable. Using the cloned genes, we showed that SIR3 at a high copy number is able to suppress mutations of SIR4. RNA blot analysis suggests that this suppression is not due to transcriptional regulation of SIR3 by SIR4; nor does any SIR4 gene transcriptionally regulate another SIR gene. Interestingly, a truncated SIR4 gene disrupts regulation of the silent mating type loci. We propose that interaction of at least the SIR3 and SIR4 gene products is involved in regulation of the silent mating type genes.

Homothallic mating type switching generates lethal chromosome breaks in rad52 strains of Saccharomyces cerevisiae

1981 ◽  
Vol 1 (6) ◽  
pp. 522-534
Author(s):  
B Weiffenbach ◽  
J E Haber
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mating Type ◽  
Deoxyribonucleic Acid ◽  
Type Locus ◽  
Mating Type Switching ◽  
Lethal Event ◽  
Bona Fide ◽  
Chromosome Iii ◽  
Mat Locus ◽  
Type Information

In homothallic cells of Saccharomyces cerevisiae, a or alpha mating type information at the mating type locus (MAT) is replaced by the transposition of the opposite mating type allele from HML alpha or HMRa. The rad52-1 mutation, which reduces mitotic and abolishes meiotic recombination, also affects homothallic switching (Malone and Esposito, Proc. Natl. Acad. Sci. U.S.A. 77:503-507, 1980). We have found that both HO rad52 MATa and HO rad52 MAT alpha cells die. This lethality is suppressed by mutations that substantially reduce but do not eliminate homothallic conversions. These mutations map at or near the MAT locus (MAT alpha inc, MATa-inc, MATa stk1) or are unlinked to MAT (HO-1 and swi1). These results suggest that the switching event itself is involved in the lethality. With the exception of swi1, HO rad52 strains carrying one of the above mutations cannot convert mating type at all. MAT alpha rad52 HO swi1 strains apparently can switch MAT alpha to MATa. However, when we analyzed these a maters, we found that few, if any, of them were bona fide MATa cells. These a-like cells were instead either deleted for part of chromosome III distal to and including MAT or had lost the entire third chromosome. Approximately 30% of the time, an a-like cell could be repaired to a normal MATa genotype if the cell was mated to a RAD52 MAT alpha-inc strain. The effects of rad52 were also studied in mata/MAT alpha-inc rad52/rad52 ho/HO diploids. When this diploid attempted to switch mata to MATa, an unstable broken chromosome was generated in nearly every cell. These studies suggest that homothallic switching involves the formation of a double-stranded deoxyribonucleic acid break or a structure which is labile in rad52 cells and results in a broken chromosome. We propose that the production of a double-stranded deoxyribonucleic acid break is the lethal event in rad52 HO cells.

INTERCONVERSION OF YEAST MATING TYPES II. RESTORATION OF MATING ABILITY TO STERILE MUTANTS IN HOMOTHALLIC AND HETEROTHALLIC STRAINS

Genetics ◽  
1977 ◽  
Vol 85 (3) ◽  
pp. 373A-393
Author(s):  
James B Hicks ◽  
Ira Herskowitz
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mating Type ◽  
Yeast Cells ◽  
Mating Types ◽  
Yeast Saccharomyces Cerevisiae ◽  
Type Locus ◽  
Mating Ability ◽  
Mating Type Locus ◽  
Regulatory Information ◽  
Ho Gene

ABSTRACT The two mating types of the yeast Saccharomyces cerevisiae can be interconverted in both homothallic and heterothallic strains. Previous work indicates that all yeast cells contain the information to be both a and α and that the HO gene (in homothallic strains) promotes a change in mating type by causing a change at the mating type locus itself. In both heterothallic and homothallic strains, a defective α mating type locus can be converted to a functional a locus and subsequently to a functional α locus. In contrast, action of the HO gene does not restore mating ability to a strain defective in another gene for mating which is not at the mating type locus. These observations indicate that a yeast cell contains an additional copy (or copies) of α information, and lead to the "cassette" model for mating type interconversion. In this model, HM  a and hmα loci are blocs of unexpressed α regulatory information, and HMα and hm  a loci are blocs of unexpressed a regulatory information. These blocs are silent because they lack an essential site for expression, and become active upon insertion of this information (or a copy of the information) into the mating type locus by action of the HO gene.

Functional domains of SIR4, a gene required for position effect regulation in Saccharomyces cerevisiae

1987 ◽  
Vol 7 (12) ◽  
pp. 4441-4452
Author(s):  
M Marshall ◽  
D Mahoney ◽  
A Rose ◽  
J B Hicks ◽  
J R Broach
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mating Type ◽  
Position Effect ◽  
Protein Activity ◽  
Multicomponent Complex ◽  
Mating Type Genes ◽  
Chromosome Iii ◽  
Covalently Linked ◽  
High Level ◽  
Mat Locus

The product of the Saccharomyces cerevisiae SIR4 gene, in conjunction with at least three other gene products, prevents expression of mating-type genes resident at loci at either end of chromosome III, but not of the same genes resident at the MAT locus in the middle of the chromosome. To address the mechanism of this novel position effect regulation, we have conducted a structural and genetic analysis of the SIR4 gene. We have determined the nucleotide sequence of the gene and found that it encodes a lysine-rich, serine-rich protein of 152 kilodaltons. Expression of the carboxy half of the protein complements a chromosomal nonsense mutation of sir4 but not a complete deletion of the gene. These results suggest that SIR4 protein activity resides in two portions of the molecule, but that these domains need not be covalently linked to execute their biological function. We also found that high-level expression of the carboxy domain of the protein yields dominant derepression of the silent loci. This anti-Sir activity can be reversed by increased expression of the SIR3 gene, whose product is normally also required for maintaining repression of the silent loci. These results are consistent with the hypothesis that SIR3 and SIR4 proteins physically associate to form a multicomponent complex required for repression of the silent mating-type loci.

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