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Molecular and Cellular Biology, March 2003, p. 1935-1945, Vol. 23, No. 6
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.6.1935-1945.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Evidence for DNA Translocation by the ISWI Chromatin-Remodeling Enzyme

Iestyn Whitehouse,1 Chris Stockdale,1 Andrew Flaus,1 Mark D. Szczelkun,2 and Tom Owen-Hughes1*

Division of Gene Regulation and Expression, The Wellcome Trust Biocentre, Department of Biochemistry, University of Dundee, Dundee DD1 5EH,1 Department of Biochemistry, School of Medical Sciences, University of Bristol, Bristol BS8 1TD, United Kingdom2

Received 16 September 2002/ Returned for modification 31 October 2002/ Accepted 19 December 2002

The ISWI proteins form the catalytic core of a subset of ATP-dependent chromatin-remodeling activities. Here, we studied the interaction of the ISWI protein with nucleosomal substrates. We found that the ability of nucleic acids to bind and stimulate the ATPase activity of ISWI depends on length. We also found that ISWI is able to displace triplex-forming oligonucleotides efficiently when they are introduced at sites close to a nucleosome but successively less efficiently 30 to 60 bp from its edge. The ability of ISWI to direct triplex displacement was specifically impeded by the introduction of 5- or 10-bp gaps in the 3'-5' strand between the triplex and the nucleosome. In combination, these observations suggest that ISWI is a 3'-5'-strand-specific, ATP-dependent DNA translocase that may be capable of forcing DNA over the surface of nucleosomes.

* Corresponding author. Mailing address: Division of Gene Regulation and Expression, The Wellcome Trust Biocentre, Department of Biochemistry, University of Dundee, Dundee DD1 5EH, United Kingdom. Phone: (44) 1382-345796. Fax: (44) 1382-348072. E-mail: t.a.owenhughes{at}dundee.ac.uk.

Molecular and Cellular Biology, March 2003, p. 1935-1945, Vol. 23, No. 6
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.6.1935-1945.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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