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Papers In Press, published online ahead of print July 14, 2004
Department of Biochemistry and Molecular Biology, Penn State University, University Park, PA 16802
Corresponding Author: jcr8{at}psu.edu
The Ssn6-Tup1 corepressor complex regulates many genes in Saccharomyces cerevisiae. Three mechanisms have been proposed to explain its repression functions: (1) nucleosome positioning by binding histone tails; (2) recruitment of histone deacetylases; (3) direct interference with the general transcription machinery or activators. It is unclear if Ssn6-Tup1 utilizes each of these mechanisms at a single gene in a redundant manner or each individually at different loci. A systematic analysis of the contribution of each mechanism at a native promoter has not been reported. Here we employed a genetic strategy to analyze the contributions of nucleosome positioning, histone deacetylation and Mediator interference in the repression of chromosomal Tup1 target genes in vivo. We exploited the fact that Ssn6-Tup1 requires the ISW2 chromatin remodeling complex to establish nucleosome positioning in vivo to disrupt chromatin structure without affecting other Tup1 repression functions. Deleting ISW2, the histone deacetylase gene HDA1 or genes encoding Mediator subunits individually caused slight or no derepression of RNR3 and HUG1. However, when Mediator mutations were combined with isw2 or hda1 mutations, enhanced transcription was observed, and the strongest level of derepression was observed in triple isw2/hda1/Mediator mutants. The increased transcription in the mutants was not due to the loss of Tup1 at the promoter, and correlated with increased TBP crosslinking to promoters. Thus, Tup1 utilizes multiple redundant mechanisms to repress transcription of native genes, which may be important for it to act as a global corepressor at a wide variety of promoters.
J. Biol. Chem, 10.1074/jbc.M407159200
Submitted on June 25, 2004
Revised on July 14, 2004
Accepted on July 14, 2004
Redundant mechanisms are used by Ssn6-Tup1 in repressing chromosomal gene transcription in saccharomyces cerevisiae
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