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Originally published In Press as doi:10.1074/jbc.M010470200 on January 24, 2001

J. Biol. Chem., Vol. 276, Issue 16, 12636-12644, April 20, 2001
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The Interactions of Yeast SWI/SNF and RSC with the Nucleosome before and after Chromatin Remodeling*

Sarojini M. SenguptaDagger , Michael VanKaneganDagger , Jim PersingerDagger , Colin Logie§, Bradley R. Cairns||, Craig L. Peterson§, and Blaine BartholomewDagger **

From the Dagger  Program in Molecular Biology, Microbiology, and Molecular Biology and Department of Biochemistry and Molecular Biology, Southern Illinois University School of Medicine, Carbondale, Illinois 62901-4413, the § Program in Molecular Medicine and Department of Biochemistry and Molecular Biology, University of Massachusetts Medical School Worcester, Massachusetts 01605, and the || Huntsman Cancer Institute and Department of Oncological Sciences, University of Utah School of Medicine, Salt Lake City, Utah 84112

Interactions of the yeast chromatin-remodeling complexes SWI/SNF and RSC with nucleosomes were probed using site-specific DNA photoaffinity labeling. 5 S rDNA was engineered with photoreactive nucleotides incorporated at different sites in DNA to scan for the subunits of SWI/SNF in close proximity to DNA when SWI/SNF is bound to the 5 S nucleosome or to the free 5 S rDNA. The Swi2/Snf2 and Snf6 subunits of SWI/SNF were efficiently cross-linked at several positions in the nucleosome, whereas only Snf6 was efficiently cross-linked when SWI/SNF was bound to free DNA. DNA photoaffinity labeling of RSC showed that the Rsc4 subunit is in close proximity to nucleosomal DNA and not when RSC is bound to free DNA. After remodeling, the Swi2/Snf2 and Rsc4 subunits are no longer detected near the nucleosomal DNA and are evidently displaced from the surface of the nucleosome, indicating significant changes in SWI/SNF and RSC contacts with DNA after remodeling.

* This work was supported by the Public Health Service Grant GM48413 from National Institutes of Health (NIGMS).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.

Present address: Nijmegen University, Moleculaire Biologie, Toernooiveld 1, 6525-EDNijmegen, The Netherlands.

** To whom correspondence should be addressed. Tel.: 618-453-6437; Fax: 618-453-6440; E-mail: bbartholomew@siumed.edu.

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


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