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J. Biol. Chem., Vol. 279, Issue 9, 7678-7684, February 27, 2004

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A Short-range Gradient of Histone H3 Acetylation and Tup1p Redistribution at the Promoter of the Saccharomyces cerevisiae SUC2 Gene^*

Abdelhalim Boukaba¶, Elena I. Georgieva¶||**, Fiona A. Myers, Alan W. Thorne, Gerardo López-Rodas, Colyn Crane-Robinson, and Luis Franco

From the Department of Biochemistry and Molecular Biology, University of Valencia, Burjassot, E-46100 Valencia, Spain, ^||Institute of Genetics, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria, and Biophysics Laboratories, Institute of Biomedical and Biomolecular Sciences, Faculty of Science, University of Portsmouth, Portsmouth PO1 2DT, United Kingdom

Chromatin immunoprecipitation assays are used to map H3 and H4 acetylation over the promoter nucleosomes and the coding region of the Saccharomyces cerevisiae SUC2 gene, under repressed and derepressed conditions, using wild type and mutant strains. In wild type cells, a high level of H3 acetylation at the distal end of the promoter drops sharply toward the proximal nucleosome that covers the TATA box, a gradient that become even steeper on derepression. In contrast, substantial H4 acetylation shows no such gradient and extends into the coding region. Overall levels of both H3 and H4 acetylation rise on derepression. Mutation of GCN5 or SNF2 lead to substantially reduced SUC2 expression; in gnc5 there is no reduction in basal H3 acetylation, but large reductions occur on derepression. SNF2 mutation has little effect on H3 acetylation, so SAGA and SWI/SNF recruitment seem to be independent events. H4 acetylation is little affected by either GCN5 or SNF2 mutation. In a double snf2/gcn5 mutant (very low SUC2 expression), H3 acetylation is at the minimal level, but H4 acetylation remains largely unaffected. Transcription is thus linked to H3 but not H4 acetylation. Chromatin immunoprecipitation assays show that Tup1p is evenly distributed over the four promoter nucleosomes in repressed wild type cells but redistributes upstream on derepression, a movement probably linked to its conversion from a repressor to an activator.

Received for publication, October 2, 2003 , and in revised form, December 10, 2003.

^* This work was supported in part by Grant BMC2001-2868 from Ministerio de Ciencia y Tecnología (Spain). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Present address: Wellcome Trust Centre for Cell Biology, ICMB, 6.33 Swann Bldg., University of Edinburgh, Mayfield Rd., Edinburgh EH9 3JR, Scotland, United Kingdom.

^¶ These two authors, listed in alphabetical order, contributed equally to this work.

^** Work at the University of Valencia was carried out during the tenure of Sabbatical Grant SAB1999-0216 from the Spanish Ministry of Educación, Cultura y Deporte.

To whom correspondence should be addressed: Dept. of Biochemistry and Molecular Biology, University of Valencia, Dr. Moliner 50, Burjassot, E-46100 Valencia, Spain. Tel.: 3496-3544869; Fax: 3496-3544635; E-mail: luis.franco{at}uv.es.

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