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PMID: 9199289 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Analysis of a meiosis-specific URS1 site: sequence requirements and involvement of replication protein A.

Molecular and cellular biology ·Vol. 17 ·No. 7 ·1997-07-00 ·Pages 3536-46

Gailus-Durner V, Chintamaneni C, Wilson R, Brill SJ, Vershon AK

Abstract

URS1 is a transcriptional repressor site found in the promoters of a wide variety of yeast genes that are induced under stress conditions. In the context of meiotic promoters, URS1 sites act as repressor sequences during mitosis and function as activator sites during meiosis. We have investigated the sequence requirements of the URS1 site of the meiosis-specific HOP1 gene (URS1H) and have found differences compared with a URS1 site from a nonmeiotic gene. We have also observed that the sequence specificity for meiotic activation at this site differs from that for mitotic repression. Base pairs flanking the conserved core sequence enhance meiotic induction but are not required for mitotic repression of HOP1. Electrophoretic mobility shift assays of mitotic and meiotic cell extracts show a complex pattern of DNA-protein complexes, suggesting that several different protein factors bind specifically to the site. We have determined that one of the complexes of URS1H is formed by replication protein A (RPA). Although RPA binds to the double-stranded URS1H site in vitro, it has much higher affinity for single-stranded than for double-stranded URS1H, and one-hybrid assays suggest that RPA does not bind to this site at detectable levels in vivo. In addition, conditional-lethal mutations in RPA were found to have no effect on URS1H-mediated repression. These results suggest that although RPA binds to URS1H in vitro, it does not appear to have a functional role in transcriptional repression through this site in vivo.

MeSH Terms
Base Sequence Binding Sites DNA, Fungal/genetics DNA-Binding Proteins/genetics,metabolism Fungal Proteins/genetics,metabolism Gene Expression Regulation, Fungal Meiosis Mitosis Mutagenesis, Site-Directed Promoter Regions, Genetic Regulatory Sequences, Nucleic Acid Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Structure-Activity Relationship
Chemicals
DNA, Fungal DNA-Binding Proteins Fungal Proteins HOP1 protein, S cerevisiae Saccharomyces cerevisiae Proteins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Gailus-Durner V
Waksman Institute of Microbiology, Rutgers University, Piscataway, New Jersey.
Chintamaneni C
Wilson R
Brill S J
Vershon A K
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.17.7.3536
Published
1997-07-00
Pages
3536-46
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC232207
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.17.7.3536 PMC Full Text View journal details
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