scholarly journals Ty insertions at two loci account for most of the spontaneous antimycin A resistance mutations during growth at 15 degrees C of Saccharomyces cerevisiae strains lacking ADH1.

1986 ◽  
Vol 6 (1) ◽  
pp. 70-79 ◽  
Author(s):  
C E Paquin ◽  
V M Williamson
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Alcohol Dehydrogenase ◽  
Transposable Element ◽  
Structural Gene ◽  
Antimycin A ◽  
Alpha Cells ◽  
Spontaneous Mutations ◽  
Resistance Mutations ◽  
Transposition Rate ◽  
Functional Copy

The mutation rate to antimycin A resistance was determined for strains of Sacchromyces cerevisiae lacking a functional copy of the structural gene for alcohol dehydrogenase I (ADH1). One type of mutation that can cause antimycin A resistance in these strains is insertion of the transposable element Ty 5' to ADH2, the structural gene for the glucose-repressed isozyme of alcohol dehydrogenase, resulting in expression of this gene during growth on glucose. Here we show that after growth at 15 or 20 degrees C on glucose, 30% of the antimycin A resistance mutations are Ty insertions at ADH2 and another 65% of the mutations are Ty insertions at ADH4, a new locus identified and cloned as described in this paper. At 30 degrees C only 6% of the mutations are Ty insertions at either of these two loci. In addition, we show that the transposition rate is lower in mating-incompetent (a/alpha) cells than in either haploid or diploid mating-competent cells. Our results suggest that under certain conditions Ty transposition may be a major cause of spontaneous mutations in S. cerevisiae.

Ty insertions at two loci account for most of the spontaneous antimycin A resistance mutations during growth at 15 degrees C of Saccharomyces cerevisiae strains lacking ADH1

1986 ◽  
Vol 6 (1) ◽  
pp. 70-79
Author(s):  
C E Paquin ◽  
V M Williamson
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Alcohol Dehydrogenase ◽  
Transposable Element ◽  
Structural Gene ◽  
Antimycin A ◽  
Alpha Cells ◽  
Spontaneous Mutations ◽  
Resistance Mutations ◽  
Transposition Rate ◽  
Functional Copy

The mutation rate to antimycin A resistance was determined for strains of Sacchromyces cerevisiae lacking a functional copy of the structural gene for alcohol dehydrogenase I (ADH1). One type of mutation that can cause antimycin A resistance in these strains is insertion of the transposable element Ty 5' to ADH2, the structural gene for the glucose-repressed isozyme of alcohol dehydrogenase, resulting in expression of this gene during growth on glucose. Here we show that after growth at 15 or 20 degrees C on glucose, 30% of the antimycin A resistance mutations are Ty insertions at ADH2 and another 65% of the mutations are Ty insertions at ADH4, a new locus identified and cloned as described in this paper. At 30 degrees C only 6% of the mutations are Ty insertions at either of these two loci. In addition, we show that the transposition rate is lower in mating-incompetent (a/alpha) cells than in either haploid or diploid mating-competent cells. Our results suggest that under certain conditions Ty transposition may be a major cause of spontaneous mutations in S. cerevisiae.

scholarly journals Physical evidence for a Saccharomyces cerevisiae transposable element which carries the his4C gene.

1981 ◽  
Vol 1 (4) ◽  
pp. 381-386 ◽  
Author(s):  
F de Bruijn ◽  
H Greer
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Transposable Element ◽  
Structural Gene ◽  
Deoxyribonucleic Acid ◽  
Physical Evidence ◽  
Precise Excision ◽  
Mutator Activity ◽  
New Location

A Saccharomyces cerevisiae transposable element which carries the his4C structural gene and which is capable of transposition, excision, and mutator activity is described. Physical evidence is presented for transposition of the his4C deoxyribonucleic acid sequences to a new location in the genome and for precise excision of these transposed deoxyribonucleic acid sequences in spontaneous his4C- segregants.

scholarly journals Alpha-factor structural gene mutations in Saccharomyces cerevisiae: effects on alpha-factor production and mating.

1985 ◽  
Vol 5 (4) ◽  
pp. 787-796 ◽  
Author(s):  
J Kurjan
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Structural Gene ◽  
Gene Mutations ◽  
Alpha Cells ◽  
Structural Genes ◽  
Coding Region ◽  
Considerable Decrease ◽  
Factor Production ◽  
Alpha Factor

The role of alpha-factor structural genes MF alpha 1 and MF alpha 2 in alpha-factor production and mating has been investigated by the construction of mf alpha 1 and mf alpha 2 mutations that totally eliminate gene function. An mf alpha 1 mutant in which the entire coding region is deleted shows a considerable decrease in alpha-factor production and a 75% decrease in mating. Mutations in mf alpha 2 have little or no effect on alpha-factor production or mating. The mf alpha 1 mf alpha 2 double mutants are completely defective in mating and alpha-factor production. These results indicate that at least one alpha-factor structural gene product is required for mating in MAT alpha cells, that MF alpha 1 is responsible for the majority of alpha-factor production, and that MF alpha 1 and MF alpha 2 are the only active alpha-factor genes.

Alpha-factor structural gene mutations in Saccharomyces cerevisiae: effects on alpha-factor production and mating

1985 ◽  
Vol 5 (4) ◽  
pp. 787-796
Author(s):  
J Kurjan
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Structural Gene ◽  
Gene Mutations ◽  
Alpha Cells ◽  
Structural Genes ◽  
Coding Region ◽  
Considerable Decrease ◽  
Factor Production ◽  
Alpha Factor

The role of alpha-factor structural genes MF alpha 1 and MF alpha 2 in alpha-factor production and mating has been investigated by the construction of mf alpha 1 and mf alpha 2 mutations that totally eliminate gene function. An mf alpha 1 mutant in which the entire coding region is deleted shows a considerable decrease in alpha-factor production and a 75% decrease in mating. Mutations in mf alpha 2 have little or no effect on alpha-factor production or mating. The mf alpha 1 mf alpha 2 double mutants are completely defective in mating and alpha-factor production. These results indicate that at least one alpha-factor structural gene product is required for mating in MAT alpha cells, that MF alpha 1 is responsible for the majority of alpha-factor production, and that MF alpha 1 and MF alpha 2 are the only active alpha-factor genes.

Physical evidence for a Saccharomyces cerevisiae transposable element which carries the his4C gene

1981 ◽  
Vol 1 (4) ◽  
pp. 381-386
Author(s):  
F de Bruijn ◽  
H Greer
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Transposable Element ◽  
Structural Gene ◽  
Deoxyribonucleic Acid ◽  
Physical Evidence ◽  
Precise Excision ◽  
Mutator Activity ◽  
New Location

A Saccharomyces cerevisiae transposable element which carries the his4C structural gene and which is capable of transposition, excision, and mutator activity is described. Physical evidence is presented for transposition of the his4C deoxyribonucleic acid sequences to a new location in the genome and for precise excision of these transposed deoxyribonucleic acid sequences in spontaneous his4C- segregants.

Carbon source dependence of transposable element-associated gene activation in Saccharomyces cerevisiae

1984 ◽  
Vol 4 (1) ◽  
pp. 61-68
Author(s):  
A K Taguchi ◽  
M Ciriacy ◽  
E T Young
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Carbon Source ◽  
Transposable Element ◽  
Mating Type ◽  
Gene Activation ◽  
Regulatory Region ◽  
Alpha Cells ◽  
Dehydrogenase Isozyme ◽  
Nonfermentable Carbon Source ◽  
Flanking Region

Seven cis-dominant mutations leading to the overproduction of the glucose-repressible alcohol dehydrogenase isozyme ADHII (structural gene, ADH2) in Saccharomyces cerevisiae have previously been shown to be due to insertion of a transposable element, Ty, in the 5' regulatory region of the ADH2 gene. We showed that although mating-competent cells (a, alpha, a/a, or alpha/alpha cells) overproduced both ADHII enzyme and ADH2 mRNA, mating-incompetent cells (a/alpha or ste-cells) produced much less ADHII enzyme and ADH2 mRNA. This mating type effect on ADH2 expression was greatest in the presence of a normally derepressing carbon source, glycerol, and much less apparent in the presence of a repressing carbon source, glucose. In addition, Ty insertion led to an aberrant carbon source response in mating-incompetent cells--the normally glucose-repressible ADHII becomes glycerol repressible. The mating type effect and aberrant carbon source response in mating-incompetent cells was specific for Ty-associated mutations in the 5' flanking region of the ADH2 gene in that a non-Ty mutation in the same region did not show these effects. Finally, Ty1 RNA levels also showed a/alpha, suppression, which was apparent only during growth on a nonfermentable carbon source such as glycerol. This suggests that Ty-mediated gene expression is subject to regulation by both mating competence and carbon catabolites.

scholarly journals Carbon source dependence of transposable element-associated gene activation in Saccharomyces cerevisiae.

1984 ◽  
Vol 4 (1) ◽  
pp. 61-68 ◽  
Author(s):  
A K Taguchi ◽  
M Ciriacy ◽  
E T Young
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Carbon Source ◽  
Transposable Element ◽  
Mating Type ◽  
Gene Activation ◽  
Regulatory Region ◽  
Alpha Cells ◽  
Dehydrogenase Isozyme ◽  
Nonfermentable Carbon Source ◽  
Flanking Region

Seven cis-dominant mutations leading to the overproduction of the glucose-repressible alcohol dehydrogenase isozyme ADHII (structural gene, ADH2) in Saccharomyces cerevisiae have previously been shown to be due to insertion of a transposable element, Ty, in the 5' regulatory region of the ADH2 gene. We showed that although mating-competent cells (a, alpha, a/a, or alpha/alpha cells) overproduced both ADHII enzyme and ADH2 mRNA, mating-incompetent cells (a/alpha or ste-cells) produced much less ADHII enzyme and ADH2 mRNA. This mating type effect on ADH2 expression was greatest in the presence of a normally derepressing carbon source, glycerol, and much less apparent in the presence of a repressing carbon source, glucose. In addition, Ty insertion led to an aberrant carbon source response in mating-incompetent cells--the normally glucose-repressible ADHII becomes glycerol repressible. The mating type effect and aberrant carbon source response in mating-incompetent cells was specific for Ty-associated mutations in the 5' flanking region of the ADH2 gene in that a non-Ty mutation in the same region did not show these effects. Finally, Ty1 RNA levels also showed a/alpha, suppression, which was apparent only during growth on a nonfermentable carbon source such as glycerol. This suggests that Ty-mediated gene expression is subject to regulation by both mating competence and carbon catabolites.

Protease B of Saccharomyces cerevisiae: Isolation and Regulation of the PRB1 Structural Gene

Genetics ◽  
1987 ◽  
Vol 115 (2) ◽  
pp. 255-263 ◽  
Author(s):  
Charles M Moehle ◽  
Martha W Aynardi ◽  
Michael R Kolodny ◽  
Frances J Park ◽  
Elizabeth W Jones
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Molecular Weight ◽  
Structural Gene ◽  
Genomic Library ◽  
Time Lag ◽  
Proteolytic Processing ◽  
Coding Region ◽  
Protease B ◽  
Endonuclease Mapping ◽  
Protein Molecular Weight

ABSTRACT We have isolated the structural gene, PRB1, for the vacuolar protease B of Saccharomyces cerevisiae from a genomic library by complementation of the prb1-1122 mutation. Deletion analysis localized the complementing activity to a 3.2-kilobase pair XhoI-HindIII restriction enzyme fragment. The fragment was used to identify a 2.3-kilobase mRNA. S1 endonuclease mapping indicated that the mRNA and the gene were colinear. No introns were detected. The mRNA is of sufficient size to encode a protein of about 69,000 molecular weight, a number much larger than either the mature enzyme (≃30,000 protein molecular weight) or the sole reported precursor (≃39,000 protein molecular weight). These results suggest that proteolytic processing steps beyond that thought to be catalyzed by protease A may be required to convert the initial glycosylated translation product into mature protease B. The PRB1 mRNA is made in substantial amounts only when the cells have exhausted the glucose supply and enter the diauxic plateau. There is an extended time lag between PRB1 transcription and expression of protease B activity. A deletion that removes about 83% of the coding region was constructed as a diploid heterozygote. Spores bearing the deletion germinate, grow at normal rates into colonies, and have no obvious phenotype beyond protease B deficiency.

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