GSBS Student Publications

Title

Role of the Saccharomyces cerevisiae general regulatory factor CP1 in methionine biosynthetic gene transcription

UMMS Affiliation

Graduate School of Biomedical Sciences; Department of Molecular Genetics and Microbiology

Date

4-1-1995

Document Type

Article

Medical Subject Headings

Base Sequence; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; Chromatin; Chromosomes, Fungal; Cysteine Synthase; DNA-Binding Proteins; Fungal Proteins; Gene Expression Regulation, Fungal; Genes, Reporter; Methionine; Molecular Sequence Data; Mutation; Oxidoreductases; Promoter Regions (Genetics); Protein Kinases; Saccharomyces cerevisiae; *Saccharomyces cerevisiae Proteins; *Transcription, Genetic

Disciplines

Life Sciences | Medicine and Health Sciences

Abstract

Saccharomyces cerevisiae general regulatory factor CP1 (encoded by the gene CEP1) is required for optimal chromosome segregation and methionine prototrophy. MET16-CYC1-lacZ reporter constructs were used to show that MET16 5'-flanking DNA contains a CP1-dependent upstream activation sequence (UAS). Activity of the UAS required an intact CP1-binding site, and the effects of cis-acting mutations on CP1 binding and UAS activity correlated. In most respects, MET16-CYC1-lacZ reporter gene expression mirrored that of chromosomal MET16; however, the endogenous gene was found to be activated in response to amino acid starvation (general control). The latter mechanism was both GCN4 and CP1 dependent. MET25 was also found to be activated by GCN4, albeit weakly. More importantly, MET25 transcription was strongly CP1 dependent in gcn4 backgrounds. The modulation of MET gene expression by GCN4 can explain discrepancies in the literature regarding CP1 dependence of MET gene transcription. Lastly, micrococcal nuclease digestion and indirect end labeling were used to analyze the chromatin structure of the MET16 locus in wild-type and cep1 cells. The results indicated that CP1 plays no major role in configuring chromatin structure in this region, although localized CP1-specific differences in nuclease sensitivity were detected.

Rights and Permissions

Citation: Mol Cell Biol. 1995 Apr;15(4):1879-88.

Related Resources

Link to article in PubMed

Journal Title

Molecular and cellular biology

PubMed ID

7891681