The overexpression of the CDC25 gene of Saccharomyces cerevisiae causes a derepression of GAL system and an increase of GAL4 transcription.
The CDC25 gene product is an exchange factor for Ras proteins and it activates the Ras/cAMP pathway in the yeast Saccharomyces cerevisiae. The overexpression of the CDC25 gene in S. cerevisiae cells causes a partial glucose-derepressed phenotype which is particularly evident for expression of invertase. To define domains of Cdc25 protein relevant for this derepression and to test another glucose repressed system, different to invertase, we have overexpressed different regions of the CDC25 gene under the control of a GAL-promoter. We found that a derepression of both GAL regulated promoters and invertase was related to the overexpression of CDC25 regions that contain a functional guanine nucleotide exchange (GEF) domain. The effect on GAL-promoters was particular evident when the CDC25 gene was under the control of a UASgal element and operates at transcriptional level, although a moderate derepression was found also for UASgal/lacZ reporter gene. Finally, the overexpression of the GEF domain of CDC25 also caused an increase in the expression of the GAL4 regulatory gene, while a constitutive activation of the Ras/cAMP pathway did not produce any increase in GAL4 expression. These findings indicate that the overexpression of the catalytic domain of CDC25 gene is necessary and sufficient to give a glucose-derepression of GAL promoters and of invertase. They also suggest that the derepression of GAL promoters occurs through an increase of GAL4 expression in a Ras cAMP independent way.
Rudoni
S
S
Dipartimento di Fisiologia e Biochimica Generali, Università de Milano, Milan, Italy.
Mauri
I
I
Ceriani
M
M
Coccetti
P
P
Martegani
E
E
D018797:Cell Cycle Proteins / Q000235:genetics*; D004268:DNA-Binding Proteins; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism*; D005690:Galactose / Q000378:metabolism / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D005800:Genes, Fungal; D017930:Genes, Reporter; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology; D006026:Glycoside Hydrolases / Q000378:metabolism; D007763:Lac Operon; D011401:Promoter Regions, Genetic / Q000187:drug effects; D012331:RNA, Fungal / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism*; D043324:beta-Fructofuranosidase; D020666:ras-GRF1 / Q000235:genetics*
D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C052130:GAL4 protein, S cerevisiae; D012331:RNA, Fungal; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D020666:ras-GRF1; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase; D005947:Glucose; D005690:Galactose