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PMID: 12727884 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Regions of GAL4 critical for binding to a promoter in vivo revealed by a visual DNA-binding analysis.

The EMBO journal ·Vol. 22 ·No. 9 ·2003-05-01 ·Pages 2178-87

Mizutani A, Tanaka M

Abstract

Binding of transcriptional activators to specific sites on DNA, mediated by their DNA-binding domain, is a key regulatory point in transcriptional regulation. With a GFP-based microscopic assay, we investigated how the prototypical activator GAL4 effectively binds to a promoter in living yeast cells. We show that GAL4 relies on a previously unrevealed mechanism involving 'DNA-binding enhancers' (DBEs), the regions of GAL4 that assist DNA-binding domain association with DNA. GAL4 contains two DBEs, one, but not the other, physically overlapping the principal transcriptional activation domain. Either of the DBEs, however, can function independently of transcriptional activation, indicating a discrete mechanism responsible for DNA-binding enhancement. The effect of DBEs, while not limited to natural target promoters, is still not universal and can be profoundly affected by the binding-site context. The GAL4 DBEs can also enhance promoter binding of an unrelated DNA-binding domain, and possibly represent a new modular functional unit responsible for effective binding of diverse regulatory factors to DNA in vivo.

MeSH Terms
Amino Acid Sequence Base Sequence DNA Primers DNA, Fungal/metabolism DNA-Binding Proteins Green Fluorescent Proteins Luminescent Proteins/metabolism Molecular Sequence Data Promoter Regions, Genetic Protein Binding Saccharomyces cerevisiae/metabolism Saccharomyces cerevisiae Proteins/metabolism Transcription Factors/metabolism
Chemicals
DNA Primers DNA, Fungal DNA-Binding Proteins GAL4 protein, S cerevisiae Luminescent Proteins Saccharomyces cerevisiae Proteins Transcription Factors Green Fluorescent Proteins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Mizutani Akiko
Unit Process and Combined Circuit, PRESTO, Japan Science and Technology Corporation (JST), Kawaguchi, Saitama 332-0012, Japan.
Tanaka Masafumi
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/cdg220
Published
2003-05-01
Pages
2178-87
Language
English
Region
England
NLM ID
8208664
PMCID
PMC156092
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/cdg220 PMC Full Text View journal details
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