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PMID: 9154826 Published · ppublish English Journal Article

Evidence that complex formation by Bas1p and Bas2p (Pho2p) unmasks the activation function of Bas1p in an adenine-repressible step of ADE gene transcription.

Molecular and cellular biology ·Vol. 17 ·No. 6 ·1997-06-00 ·Pages 3272-83

Zhang F, Kirouac M, Zhu N, Hinnebusch AG, Rolfes RJ

Abstract

Bas1p and Bas2p (Pho2p) are Myb-related and homeodomain DNA binding proteins, respectively, required for transcription of adenine biosynthetic genes in Saccharomyces cerevisiae. The repression of ADE genes in adenine-replete cells involves down-regulation of the functions of one or both of these activator proteins. A LexA-Bas2p fusion protein was found to activate transcription from a lexAop-lacZ reporter independently of both BAS1 function and the adenine levels in the medium. In contrast, a LexA-Bas1p fusion activated the lexAop reporter in a BAS2-dependent and adenine-regulated fashion. The DNA binding activity of Bas2p was not needed for its ability to support activation of the lexAop reporter by LexA-Bas1p, indicating that LexA-Bas1p recruits Bas2p to this promoter. The activation functions of both authentic Bas1p and LexA-Bas1p were stimulated under adenine-repressing conditions by overexpression of Bas2p, suggesting that complex formation by these proteins is inhibited in adenine-replete cells. Replacement of Asp-617 with Asn in Bas1p or LexA-Bas1p allowed either protein to activate transcription under repressing conditions in a manner fully dependent on Bas2p, suggesting that this mutation reduces the negative effect of adenine on complex formation by Bas1p and Bas2p. Deletions of N-terminal and C-terminal segments from the Bas1p moiety of LexA-Bas1p allowed high-level activation by the truncated proteins independently of Bas2p and adenine levels in the medium. From these results we propose that complex formation between Bas1p and Bas2p unmasks a latent activation function in Bas1p as a critical adenine-regulated step in transcription of the ADE genes.

MeSH Terms
Adenine/biosynthesis,pharmacology,physiology Aspartic Acid Bacterial Proteins/genetics,metabolism Binding Sites DNA, Fungal/metabolism Down-Regulation/drug effects Fungal Proteins/chemistry,genetics,metabolism Homeodomain Proteins Macromolecular Substances Models, Molecular Mutagenesis, Site-Directed Repressor Proteins/genetics,metabolism Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Serine Endopeptidases/genetics,metabolism Trans-Activators/chemistry,genetics,metabolism Transcription, Genetic Transcriptional Activation
Chemicals
BAS1 protein, S cerevisiae Bacterial Proteins DNA, Fungal Fungal Proteins Homeodomain Proteins LexA protein, Bacteria Macromolecular Substances PHO2 protein, S cerevisiae Repressor Proteins Saccharomyces cerevisiae Proteins Trans-Activators Aspartic Acid Serine Endopeptidases Adenine
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Zhang F
Laboratory of Eukaryotic Gene Regulation, National Institute of Child Health and Human Development, Bethesda, Maryland 20892, USA.
Kirouac M
Zhu N
Hinnebusch A G
Rolfes R J
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.17.6.3272
Published
1997-06-00
Pages
3272-83
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC232180
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.17.6.3272 PMC Full Text View journal details
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