Overlapping but Distinct Patterns of Histone Acetylation by the Human Coactivators p300 and PCAF within Nucleosomal Substrates

R. Louis Schiltz; Craig A. Mizzen; Alex Vassilev; Richard G. Cook; C. David Allis; Yoshihiro Nakatani

doi:10.1074/jbc.274.3.1189

Overlapping but Distinct Patterns of Histone Acetylation by the Human Coactivators p300 and PCAF within Nucleosomal Substrates*

From the ^‡Laboratory of Molecular Growth Regulation, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892, the ^¶Department of Biology, University of Rochester, Rochester, New York 14627, and the ^**Department of Microbiology and Immunology, Baylor College of Medicine, Houston, Texas 77030

Abstract

A number of transcriptional coactivators possess intrinsic histone acetylase activity, providing a direct link between hyperacetylated chromatin and transcriptional activation. We have determined the core histone residues acetylated in vitro by recombinant p300 and PCAF within mononucleosomes. p300 specifically acetylates all sites of histones H2A and H2B known to be acetylated in bulk chromatin in vivo but preferentially acetylates lysines 14 and 18 of histone H3 and lysines 5 and 8 of histone H4. PCAF primarily acetylates lysine 14 of H3 but also less efficiently acetylates lysine 8 of H4. PCAF in its native form, which is present in a stable multimeric protein complex lacking p300/CBP, primarily acetylates H3 to a monoacetylated form, suggesting that PCAF-associated polypeptides do not alter the substrate specificity. These distinct patterns of acetylation by the p300 and PCAF may contribute to their differential roles in transcriptional regulation.

Footnotes

↵* The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵§ These authors contributed equally to this work.
↵‖ Present address: Dept. of Biochemistry and Molecular Genetics, University of Virginia Health Sciences Center, Charlottesville, VA 22908.
↵‡ To whom correspondence should be addressed: NIH, Bldg. 6, Rm. 416, Bethesda, MD 20892-2753. Tel.: 301-402-4904; Fax: 301-496-7823; E-mail: yoshi{at}helix.nih.gov.