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Molecular and Cellular Biology, January 2003, p. 80-91, Vol. 23, No. 1
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.1.80-91.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Yeast Isw1p Forms Two Separable Complexes In Vivo

Jay C. Vary, Jr.,1,2 Vamsi K. Gangaraju,3 Jun Qin,4,1 Carolyn Church Landel,1,2 Charles Kooperberg,5 Blaine Bartholomew,3 and Toshio Tsukiyama1*

Division of Basic Sciences,1 Division of Public Health Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109,5 Molecular and Cellular Biology Program, Fred Hutchinson Cancer Research Center and University of Washington, Seattle, Washington 98195,2 Department of Biochemistry and Molecular Biology, Southern Illinois University School of Medicine, Carbondale, Illinois 62901,3 Laboratory of Biophysical Chemistry, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 208924

Received 12 June 2002/ Returned for modification 12 August 2002/ Accepted 7 October 2002

There are several classes of ATP-dependent chromatin remodeling complexes, which modulate the structure of chromatin to regulate a variety of cellular processes. The budding yeast, Saccharomyces cerevisiae, encodes two ATPases of the ISWI class, Isw1p and Isw2p. Previously Isw1p was shown to copurify with three other proteins. Here we identify these associated proteins and show that Isw1p forms two separable complexes in vivo (designated Isw1a and Isw1b). Biochemical assays revealed that while both have equivalent nucleosome-stimulated ATPase activities, Isw1a and Isw1b differ in their abilities to bind to DNA and nucleosomal substrates, which possibly accounts for differences in specific activities in nucleosomal spacing and sliding. In vivo, the two Isw1 complexes have overlapping functions in transcriptional regulation of some genes yet distinct functions at others. In addition, these complexes show different contributions to cell growth at elevated temperatures.

* Corresponding author. Mailing address: Division of Basic Sciences, Fred Hutchinson Cancer Research Center, P.O. Box 19024, 1100 Fairview Ave. N, Mailstop A1-162, Seattle, WA 98109. Phone: (206) 667-4996. Fax: (206) 667-6497. E-mail: ttsukiya{at}fhcrc.org.

{dagger} Present address: Verna and Marrs McLean Department of Biochemistry and Molecular Biology and Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX 77030.

{ddagger} Present address: Washington Association for Biomedical Research, Seattle, WA 98121.

Molecular and Cellular Biology, January 2003, p. 80-91, Vol. 23, No. 1
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.1.80-91.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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