scholarly journals Cloning and genetic mapping of SNF1, a gene required for expression of glucose-repressible genes in Saccharomyces cerevisiae.

1984 ◽  
Vol 4 (1) ◽  
pp. 49-53 ◽  
Author(s):  
J L Celenza ◽  
M Carlson
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Genetic Mapping ◽  
Gene Product ◽  
Structural Gene ◽  
Glucose Repression ◽  
Carbon Sources ◽  
Recombinant Plasmids ◽  
The Right ◽  
Integrating Vector

A functional SNF1 gene product is required to derepress expression of many glucose-repressible genes in Saccharomyces cerevisiae. Strains carrying a snf1 mutation are unable to grow on sucrose, galactose, maltose, melibiose, or nonfermentable carbon sources; utilization of these carbon sources is regulated by glucose repression. The inability of snf1 mutants to utilize sucrose results from failure to derepress expression of the structural gene for invertase at the RNA level. We isolated recombinant plasmids carrying the SNF1 gene by complementation of the snf1 defect in S. cerevisiae. A 3.5-kilobase region is common to the DNA segments cloned in five different plasmids. Transformation of S. cerevisiae with an integrating vector carrying a segment of the cloned DNA resulted in integration of the plasmid at the SNF1 locus. This result indicates that the cloned DNA is homologous to sequences at the SNF1 locus. By mapping a plasmid marker linked to SNF1 in this transformant, we showed that the SNF1 gene is located on chromosome IV. We then mapped snf1 to a position 5.6 centimorgans distal to rna3 on the right arm; snf1 is not extremely closely linked to any previously mapped mutation.

Cloning and genetic mapping of SNF1, a gene required for expression of glucose-repressible genes in Saccharomyces cerevisiae

1984 ◽  
Vol 4 (1) ◽  
pp. 49-53
Author(s):  
J L Celenza ◽  
M Carlson
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Genetic Mapping ◽  
Gene Product ◽  
Structural Gene ◽  
Glucose Repression ◽  
Carbon Sources ◽  
Recombinant Plasmids ◽  
The Right ◽  
Integrating Vector

A functional SNF1 gene product is required to derepress expression of many glucose-repressible genes in Saccharomyces cerevisiae. Strains carrying a snf1 mutation are unable to grow on sucrose, galactose, maltose, melibiose, or nonfermentable carbon sources; utilization of these carbon sources is regulated by glucose repression. The inability of snf1 mutants to utilize sucrose results from failure to derepress expression of the structural gene for invertase at the RNA level. We isolated recombinant plasmids carrying the SNF1 gene by complementation of the snf1 defect in S. cerevisiae. A 3.5-kilobase region is common to the DNA segments cloned in five different plasmids. Transformation of S. cerevisiae with an integrating vector carrying a segment of the cloned DNA resulted in integration of the plasmid at the SNF1 locus. This result indicates that the cloned DNA is homologous to sequences at the SNF1 locus. By mapping a plasmid marker linked to SNF1 in this transformant, we showed that the SNF1 gene is located on chromosome IV. We then mapped snf1 to a position 5.6 centimorgans distal to rna3 on the right arm; snf1 is not extremely closely linked to any previously mapped mutation.

scholarly journals Multicopy FZF1 (SUL1) Suppresses the Sulfite Sensitivity but Not the Glucose Derepression or Aberrant Cell Morphology of a grr1 Mutant of Saccharomyces cerevisiae

Genetics ◽  
1996 ◽  
Vol 144 (2) ◽  
pp. 511-521 ◽  
Author(s):  
Dorina Avram ◽  
Alan T Bakalinsky
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Transcription Factor ◽  
Glucose Metabolism ◽  
Cell Morphology ◽  
Glucose Repression ◽  
Carbon Sources ◽  
Single Copy ◽  
Aberrant Cell ◽  
Growth Defects ◽  
Putative Transcription Factor

Abstract An ssu2 mutation in Sacccharomyces cermisiae, previously shown to cause sulfite sensitivity, was found to be allelic to GRR1, a gene previously implicated in glucose repression. The suppressor rgt1, which suppresses the growth defects of grr1 strains on glucose, did not fully suppress the sensitivity on glucose or nonglucose carbon sources, indicating that it is not strictly linked to a defect in glucose metabolism. Because the Cln1 protein was previously shown to be elevated in grr1 mutants, the effect of CLN1 overexpression on sulfite sensitivity was investigated. Overexpression in GRR1 cells resulted in sulfite sensitivity, suggesting a connection between CLN1 and sulfite metabolism. Multicopy FZF1, a putative transcription factor, was found to suppress the sulfite sensitive phenotype of grr1 strains, but not the glucose derepression or aberrant cell morphology. Multicopy FZF1 was also found to suppress the sensitivity of a number of other unrelated sulfite-sensitive mutants, but not that of ssu1 or met20, implying that FZF1 may act through Ssulp and Met20p. Disruption of FZF1 resulted in sulfite sensitivity when the construct was introduced in single copy at the FZF1 locus in a GRR1 strain, providing evidence that FZF1 is involved in sulfite metabolism.

scholarly journals Duplicate upstream activating sequences in the promoter region of the Saccharomyces cerevisiae GAL7 gene.

1986 ◽  
Vol 6 (1) ◽  
pp. 246-256 ◽  
Author(s):  
M Tajima ◽  
Y Nogi ◽  
T Fukasawa
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Glucose Repression ◽  
Rotational Symmetry ◽  
Consensus Sequence ◽  
Carbon Sources ◽  
Constitutive Expression ◽  
Tata Box ◽  
Base Pairs ◽  
Multicopy Vector ◽  
In Cis

We constructed a series of deletions in the 5' noncoding region of the Saccharomyces cerevisiae GAL7 gene, fused them to the Escherichia coli gene lacZ, and introduced them into yeasts by using a multicopy vector. We then studied the effect of the deletions on beta-galactosidase synthesis directed by the gene fusions in media with various carbon sources. This analysis identified a TATA box and two upstream activating sequences as necessary elements for galactose-controlled GAL7 transcription. Two upstream activating sequences exhibiting 71% homology with each other were located 255 and 168 base pairs, respectively, upstream of the GAL7 transcription start point. Each sequence consists of 21 base pairs, displaying an approximate rotational symmetry with a core consensus sequence of GAA--AGCTGCTTC--CGCG. At least one of the two sequences is required for galactose induction and also for glucose repression of the GAL7'-lac'Z gene. Analysis with host regulatory mutants delta gal14 and delta gal180 suggests that these sequences are the site at which the GAL4 product exerts its action to activate the GAL7 gene. We also observed that a deletion lacking both upstream activation sequences allowed the gene fusion to be expressed in the absence of galactose at about 10% of the fully induced level of the intact fusion. This constitutive expression depended on the presence of the TATA box of GAL7 in cis but not on a functional GAL4 gene. The level of the uncontrolled expression was decreased by increasing the distance between the TATA box and the pBR322 sequence in the vector plasmid.

scholarly journals A common bacterial metabolite elicits prion-based bypass of glucose repression

eLife ◽  
2016 ◽  
Vol 5 ◽  
Author(s):  
David M Garcia ◽  
David Dietrich ◽  
Jon Clardy ◽  
Daniel F Jarosz
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Lactic Acid ◽  
Glucose Metabolism ◽  
Carbon Source ◽  
Glucose Repression ◽  
Carbon Sources ◽  
Yeast Cells ◽  
Genetic Element ◽  
Yeast Saccharomyces Cerevisiae ◽  
The Common

Robust preference for fermentative glucose metabolism has motivated domestication of the budding yeast Saccharomyces cerevisiae. This program can be circumvented by a protein-based genetic element, the [GAR+] prion, permitting simultaneous metabolism of glucose and other carbon sources. Diverse bacteria can elicit yeast cells to acquire [GAR+], although the molecular details of this interaction remain unknown. Here we identify the common bacterial metabolite lactic acid as a strong [GAR+] inducer. Transient exposure to lactic acid caused yeast cells to heritably circumvent glucose repression. This trait had the defining genetic properties of [GAR+], and did not require utilization of lactic acid as a carbon source. Lactic acid also induced [GAR+]-like epigenetic states in fungi that diverged from S. cerevisiae ~200 million years ago, and in which glucose repression evolved independently. To our knowledge, this is the first study to uncover a bacterial metabolite with the capacity to potently induce a prion.

ABF1 is a phosphoprotein and plays a role in carbon source control of COX6 transcription in Saccharomyces cerevisiae

1992 ◽  
Vol 12 (9) ◽  
pp. 4197-4208
Author(s):  
S Silve ◽  
P R Rhode ◽  
B Coll ◽  
J Campbell ◽  
R O Poyton
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Carbon Source ◽  
Glucose Repression ◽  
Carbon Sources ◽  
Growth Conditions ◽  
Source Control ◽  
Specific Target Gene ◽  
Phosphorylation Pattern ◽  
Nonfermentable Carbon Source ◽  
In Cells

Previously, we have shown that the Saccharomyces cerevisiae DNA-binding protein ABF1 exists in at least two different electrophoretic forms (K. S. Sweder, P. R. Rhode, and J. L. Campbell, J. Biol. Chem. 263: 17270-17277, 1988). In this report, we show that these forms represent different states of phosphorylation of ABF1 and that at least four different phosphorylation states can be resolved electrophoretically. The ratios of these states to one another differ according to growth conditions and carbon source. Phosphorylation of ABF1 is therefore a regulated process. In nitrogen-starved cells or in cells grown on nonfermentable carbon sources (e.g., lactate), phosphorylated forms predominate, while in cells grown on fermentable carbon sources (e.g., glucose), dephosphorylated forms are enriched. The phosphorylation pattern is affected by mutations in the SNF1-SSN6 pathway, which is involved in glucose repression-depression. Whereas a functional SNF1 gene, which encodes a protein kinase, is not required for the phosphorylation of ABF1, a functional SSN6 gene is required for itsd ephosphorylation. The phosphorylation patterns that we have observed correlate with the regulation of a specific target gene, COX6, which encodes subunit VI of cytochrome c oxidase. Transcription of COX6 is repressed by growth in medium containing a fermentable carbon source and is derepressed by growth in medium containing a nonfermentable carbon source. COX6 repression-derepression is under the control of the SNF1-SSN6 pathway. This carbon source regulation is exerted through domain 1, a region of the upstream activation sequence UAS6 that binds ABF1 (J. D. Trawick, N. Kraut, F. Simon, and R. O. Poyton, Mol. Cell Biol. 12:2302-2314, 1992). We show that the greater the phosphorylation of ABF1, the greater the transcription of COX6. Furthermore, the ABF1-containing protein-DNA complexes formed at domain 1 differ according to the phosphorylation state of ABF1 and the carbon source on which the cells were grown. From these findings, we propose that the phosphorylation of ABF1 is involved in glucose repression-derepression of COX6 transcription.

Carbon source-dependent transcriptional regulation of the mitochondrial glycerol-3-phosphate dehydrogenase gene, GUT2, from Saccharomyces cerevisiae

2000 ◽  
Vol 46 (12) ◽  
pp. 1096-1100 ◽  
Author(s):  
Morten Grauslund ◽  
Birgitte Rønnow
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Transcriptional Regulation ◽  
Glucose Repression ◽  
Carbon Sources ◽  
Negative Regulator ◽  
Transcriptional Analysis ◽  
Dna Binding Site ◽  
Glycerol Utilization ◽  
Glycerol 3 Phosphate Dehydrogenase ◽  
Utilization Pathway

Cytosolic glycerol kinase (Gut1p) and mitochondrial glycerol-3-phosphate dehydrogenase (Gut2p) constitute the glycerol utilization pathway in Saccharomyces cerevisiae. Transcriptional analysis of the GUT2 gene showed that it was repressed by glucose and derepressed on the non-fermentable carbon sources, glycerol, lactate and ethanol. Derepression of GUT2 requires the protein kinase Snf1p as well as the heteromeric protein complex, Hap2/3/4/5, and its putative DNA-binding site (UASHAP) located in the promoter region. Furthermore, glucose repression of GUT2 requires the negative regulator, Opi1p.Key words: GUT2, mitochondrial glycerol-3-phosphate dehydrogenase, transcriptional regulation, Saccharomyces cerevisiae.

scholarly journals Structure and expression of the SNF1 gene of Saccharomyces cerevisiae.

1984 ◽  
Vol 4 (1) ◽  
pp. 54-60 ◽  
Author(s):  
J L Celenza ◽  
M Carlson
Keyword(s):  
Gene Expression ◽  
Saccharomyces Cerevisiae ◽  
Gene Product ◽  
Gene Disruption ◽  
Glucose Repression ◽  
Regulation Of Gene Expression ◽  
Null Mutation ◽  
Chromosomal Locus ◽  
Cloned Gene ◽  
External Glucose

The SNF1 gene of Saccharomyces cerevisiae is essential for normal regulation of gene expression by glucose repression. A functional SNF1 gene product is required to derepress many glucose-repressible genes in response to conditions of low external glucose. In the case of the SUC2 structural gene for invertase, SNF1 acts at the RNA level. We have reported the isolation of a cloned gene that complements the snf1 defect in S. cerevisiae and that is homologous to DNA at the SNF1 locus (J. L. Celenza and M. Carlson, Mol. Cell. Biol. 4:49-53, 1984). In this work we identified a 2.4-kilobase polyadenylate-containing RNA encoded by the SNF1 gene and showed that its level is neither regulated by glucose repression nor dependent on a functional SNF1 product. The position of the SNF1 RNA relative to the cloned DNA was mapped, and the direction of transcription was determined. The cloned DNA was used to disrupt the SNF1 gene at its chromosomal locus. Gene disruption resulted in A Snf1- phenotype, thereby proving that the cloned gene is the SNF1 gene and showing that the phenotype of a true null mutation is indistinguishable from that of previously isolated snf1 mutations.

Structure and expression of the SNF1 gene of Saccharomyces cerevisiae

1984 ◽  
Vol 4 (1) ◽  
pp. 54-60
Author(s):  
J L Celenza ◽  
M Carlson
Keyword(s):  
Gene Expression ◽  
Saccharomyces Cerevisiae ◽  
Gene Product ◽  
Gene Disruption ◽  
Glucose Repression ◽  
Regulation Of Gene Expression ◽  
Null Mutation ◽  
Chromosomal Locus ◽  
Cloned Gene ◽  
External Glucose

The SNF1 gene of Saccharomyces cerevisiae is essential for normal regulation of gene expression by glucose repression. A functional SNF1 gene product is required to derepress many glucose-repressible genes in response to conditions of low external glucose. In the case of the SUC2 structural gene for invertase, SNF1 acts at the RNA level. We have reported the isolation of a cloned gene that complements the snf1 defect in S. cerevisiae and that is homologous to DNA at the SNF1 locus (J. L. Celenza and M. Carlson, Mol. Cell. Biol. 4:49-53, 1984). In this work we identified a 2.4-kilobase polyadenylate-containing RNA encoded by the SNF1 gene and showed that its level is neither regulated by glucose repression nor dependent on a functional SNF1 product. The position of the SNF1 RNA relative to the cloned DNA was mapped, and the direction of transcription was determined. The cloned DNA was used to disrupt the SNF1 gene at its chromosomal locus. Gene disruption resulted in A Snf1- phenotype, thereby proving that the cloned gene is the SNF1 gene and showing that the phenotype of a true null mutation is indistinguishable from that of previously isolated snf1 mutations.

scholarly journals Cloning and mapping of Saccharomyces cerevisiae photoreactivation gene PHR1.

1984 ◽  
Vol 4 (9) ◽  
pp. 1864-1870 ◽  
Author(s):  
D Schild ◽  
J Johnston ◽  
C Chang ◽  
R K Mortimer
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Visible Light ◽  
Gene Product ◽  
Chromosomal Dna ◽  
Yeast Saccharomyces Cerevisiae ◽  
Pyrimidine Dimers ◽  
The Right ◽  
A Site ◽  
Primary Gene ◽  
Light Cells

The yeast Saccharomyces cerevisiae, like most organisms, is able to directly repair pyrimidine dimers by using a photoreactivating enzyme and visible light. Cells carrying the phr1 mutation were shown previously to be unable to photoreactivate dimers, but neither the map position nor the primary gene product of the PHR1 gene has been determined. We have cloned this gene and determined its map position. A plasmid containing a 6.4-kilobase yeast DNA insert has been isolated and shown to restore photoreactivation in a phr1 strain. A 3.1-kilobase subclone has also been shown to complement phr1. The original plasmid was targeted to integrate into chromosomal DNA at a site homologous to the insert by cutting within the insert. Two of these integrants have been mapped on the right arm of chromosome XV; the integrants have been further mapped at ca. 13 centimorgans from prt1. It has also been independently determined that phr1 maps at this location. Thus, we have determined the map position of PHR1 and also have shown that the plasmid contains PHR1 rather than a suppressor of the phr1 mutation.

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