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Molecular and Cellular Biology, February 2003, p. 1025-1033, Vol. 23, No. 3
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.3.1025-1033.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Interaction of Histone Acetylases and Deacetylases In Vivo

Satoshi Yamagoe,1,{dagger} Tomohiko Kanno,1 Yuka Kanno,1 Shigakazu Sasaki,1,2 Richard M. Siegel,2 Michael J. Lenardo,2 Glen Humphrey,1 Yonghong Wang,1 Yoshihiro Nakatani,3 Bruce H. Howard,1 and Keiko Ozato1*

Laboratory of Molecular Growth Regulation, National Institute of Child Health and Human Development,1 Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892,2 Dana Farber Cancer Institute and Harvard Medical School, Boston, Massachusetts 021153

Received 12 June 2002/ Returned for modification 1 August 2002/ Accepted 29 October 2002

Having opposing enzymatic activities, histone acetylases (HATs) and deacetylases affect chromatin and regulate transcription. The activities of the two enzymes are thought to be balanced in the cell by an unknown mechanism that may involve their direct interaction. Using fluorescence resonance energy transfer analysis, we demonstrated that the acetylase PCAF and histone deacetylase 1 (HDAC1) are in close spatial proximity in living cells, compatible with their physical interaction. In agreement, coimmunoprecipitation assays demonstrated that endogenous HDACs are associated with PCAF and another acetylase, GCN5, in HeLa cells. We found by glycerol gradient sedimentation analysis that HATs are integrated into a large multiprotein HDAC complex that is distinct from the previously described HDAC complexes containing mSin3A, Mi-2/NRD, or CoREST. This HDAC-HAT association is partly accounted for by a direct protein-protein interaction observed in vitro. The HDAC-HAT complex may play a role in establishing a dynamic equilibrium of the two enzymes in vivo.

* Corresponding author. Mailing address: Laboratory of Molecular Growth Regulation, National Institute of Child Health and Human Development, Bldg. 6, Rm. 2A01, National Institutes of Health, Bethesda, MD 20892-2753. Phone: (301) 496-9184. Fax: (301) 480-9354. E-mail: ozatok{at}nih.gov.

{dagger} Present address: Department of Bioactive Molecules, National Institute of Infectious Diseases, Tokyo, Japan.

{ddagger} Present address: Second Division of Internal Medicine, Hamamatsu University School of Medicine, Hamamatsu, Japan.

Molecular and Cellular Biology, February 2003, p. 1025-1033, Vol. 23, No. 3
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.3.1025-1033.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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