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PMID: 2649888 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Yeast GCN4 transcriptional activator protein interacts with RNA polymerase II in vitro.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 86 ·No. 8 ·1989-04-00 ·Pages 2652-6

Brandl CJ, Struhl K

Abstract

Regulated transcription by eukaryotic RNA polymerase II (Pol II) requires the functional interaction of multiple protein factors, some of which presumably interact directly with the polymerase. One such factor, the yeast GCN4 activator protein, binds to the upstream promoter elements of many amino acid biosynthetic genes and induces their transcription. Through the use of affinity chromatography involving GCN4- or Pol II-Sepharose columns, we show that GCN4 interacts specifically with Pol II in vitro. Purified Pol II is retained on the GCN4-Sepharose column under conditions in which the vast majority of proteins flow through. Moreover, Pol II can be selectively isolated from more complex mixtures of proteins. Conversely, GCN4 protein, synthesized in vitro or in Escherichia coli, specifically binds to the Pol II-Sepharose column under equivalent conditions. Using deletion mutants, we also show that the DNA-binding domain of GCN4 is both necessary and sufficient for this interaction. We suggest the possibility that this GCN4-Pol II interaction may be important for transcription in vivo.

MeSH Terms
Binding Sites Chromatography, Affinity DNA-Binding Proteins/metabolism Gene Expression Regulation In Vitro Techniques Protein Binding RNA Polymerase II/metabolism Saccharomyces cerevisiae/genetics Transcription Factors/metabolism Transcription, Genetic
Chemicals
DNA-Binding Proteins Transcription Factors RNA Polymerase II
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Brandl C J
Department of Biological Chemistry, Harvard Medical School, Boston, MA 02115.
Struhl K
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38 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.86.8.2652
Published
1989-04-00
Pages
2652-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC286975
Subset
IM
Grants
NIGMS NIH HHS · GM30186 · United States
External Links
PubMed source DOI: 10.1073/pnas.86.8.2652 PMC Full Text View journal details
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