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Originally published In Press as doi:10.1074/jbc.M100125200 on February 5, 2001

J. Biol. Chem., Vol. 276, Issue 18, 14773-14783, May 4, 2001
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ATP-dependent Nucleosome Remodeling and Histone Hyperacetylation Synergistically Facilitate Transcription of Chromatin*

Gaku MizuguchiDagger §, Alex Vassilev, Toshio TsukiyamaDagger ||, Yoshihiro Nakatani**, and Carl WuDagger Dagger Dagger

From the Dagger  Laboratory of Molecular Cell Biology, NCI, National Institutes of Health, Bethesda, Maryland 20892, § Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan,  Laboratory of Molecular Growth Regulation, NICHD, National Institutes of Health, Bethesda, Maryland 20892, the || Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, and ** Dana Farber Cancer Institute, Boston, Massachusetts 02115

Drosophila nucleosome remodeling factor (NURF) is an ISWI-containing protein complex that facilitates nucleosome mobility and transcriptional activation in an ATP-dependent manner. Numerous studies have implicated histone acetylation in transcriptional activation. We investigated the relative contributions of these two chromatin modifications to transcription in vitro of a chromatinized adenovirus E4 minimal promoter that contains binding sites for the GAL4-VP16 activator. We found that NURF could remodel chromatin and stimulate transcription irrespective of the acetylation status of histones. In contrast, hyperacetylation of histones in the absence of NURF was unable to stimulate transcription, suggesting that NURF-dependent chromatin remodeling is an obligatory step in E4 promoter activation. When chromatin templates were first hyperacetylated and then incubated with NURF, significantly greater transcription stimulation was observed. The results suggest that changes in chromatin induced by acetylation of histones and the mobilization of nucleosomes by NURF combine synergistically to facilitate transcription. Experiments using single and multiple rounds of transcription indicate that these chromatin modifications stimulate transcription preinitiation as well as reinitiation.

* This work was supported by the Intramural Research Program of NCI, National Institutes of Health, Division of Basic Sciences.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.

Dagger Dagger To whom correspondence should be addressed. carlwu@helix.nih.gov.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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