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PMID: 10409719 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S.

Chromatin opening and transactivator potentiation by RAP1 in Saccharomyces cerevisiae.

Molecular and cellular biology ·Vol. 19 ·No. 8 ·1999-08-00 ·Pages 5279-88

Yu L, Morse RH

Abstract

Transcriptional activators function in vivo via binding sites that may be packaged into chromatin. Here we show that whereas the transcriptional activator GAL4 is strongly able to perturb chromatin structure via a nucleosomal binding site in yeast, GCN4 does so poorly. Correspondingly, GCN4 requires assistance from an accessory protein, RAP1, for activation of the HIS4 promoter, whereas GAL4 does not. The requirement for RAP1 for GCN4-mediated HIS4 activation is dictated by the DNA-binding domain of GCN4 and not the activation domain, suggesting that RAP1 assists GCN4 in gaining access to its binding site. Consistent with this, overexpression of GCN4 partially alleviates the requirement for RAP1, whereas HIS4 activation via a weak GAL4 binding site requires RAP1. RAP1 is extremely effective at interfering with positioning of a nucleosome containing its binding site, consistent with a role in opening chromatin at the HIS4 promoter. Furthermore, increasing the spacing between binding sites for RAP1 and GCN4 by 5 or 10 bp does not impair HIS4 activation, indicating that cooperative protein-protein interactions are not involved in transcriptional facilitation by RAP1. We conclude that an important role of RAP1 is to assist activator binding by opening chromatin.

MeSH Terms
Alcohol Oxidoreductases Aminohydrolases Chromatin/metabolism,ultrastructure DNA, Fungal/genetics,metabolism DNA-Binding Proteins/physiology Drosophila Proteins Fungal Proteins/genetics,physiology Gene Expression Regulation, Fungal Homeodomain Proteins/metabolism Promoter Regions, Genetic Protein Binding Protein Kinases/physiology Pyrophosphatases Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins Trans-Activators/metabolism Transcription Factors/genetics,physiology Transcriptional Activation
Chemicals
Chromatin DNA, Fungal DNA-Binding Proteins Drosophila Proteins Fungal Proteins GAL4 protein, S cerevisiae Homeodomain Proteins Saccharomyces cerevisiae Proteins Trans-Activators Transcription Factors bcd protein, Drosophila Alcohol Oxidoreductases HIS4 protein, S cerevisiae Protein Kinases Aminohydrolases Pyrophosphatases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Yu L
Molecular Genetics Program, Wadsworth Center, New York State Department of Health, and State University of New York School of Public Health, Albany, New York 12201-2002, USA.
Morse R H
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.19.8.5279
Published
1999-08-00
Pages
5279-88
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC84371
Subset
IM
Grants
NIGMS NIH HHS · R01 GM051993 · United States
NIGMS NIH HHS · GM51993 · United States
External Links
PubMed source DOI: 10.1128/MCB.19.8.5279 PMC Full Text View journal details
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