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Originally published In Press as doi:10.1074/jbc.M006784200 on November 10, 2000

J. Biol. Chem., Vol. 276, Issue 5, 3024-3030, February 2, 2001
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Unwinding of a DNA Triple Helix by the Werner and Bloom Syndrome Helicases*

Robert M. Brosh Jr.Dagger , Alokes MajumdarDagger , Shital DesaiDagger , Ian D. Hickson§, Vilhelm A. BohrDagger , and Michael M. SeidmanDagger

From the Dagger  Laboratory of Molecular Genetics, NIA, National Institutes of Health, Baltimore, Maryland 21224 and § Imperial Cancer Research Fund Laboratories, Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DS, United Kingdom

Bloom syndrome and Werner syndrome are genome instability disorders, which result from mutations in two different genes encoding helicases. Both enzymes are members of the RecQ family of helicases, have a 3' right-arrow 5' polarity, and require a 3' single strand tail. In addition to their activity in unwinding duplex substrates, recent studies show that the two enzymes are able to unwind G2 and G4 tetraplexes, prompting speculation that failure to resolve these structures in Bloom syndrome and Werner syndrome cells may contribute to genome instability. The triple helix is another alternate DNA structure that can be formed by sequences that are widely distributed throughout the human genome. Here we show that purified Bloom and Werner helicases can unwind a DNA triple helix. The reactions are dependent on nucleoside triphosphate hydrolysis and require a free 3' tail attached to the third strand. The two enzymes unwound triplexes without requirement for a duplex extension that would form a fork at the junction of the tail and the triplex. In contrast, a duplex formed by the third strand and a complement to the triplex region was a poor substrate for both enzymes. However, the same duplex was readily unwound when a noncomplementary 5' tail was added to form a forked structure. It seems likely that structural features of the triplex mimic those of a fork and thus support efficient unwinding by the two helicases.

* 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.

To whom correspondence should be addressed: LMG, Box 1, GRC, 5600 Nathan Shock Dr., Baltimore, MD 21224. Tel.: 410-558-8565; Fax: 410-558-8157; E-mail: seidmanm@grc.nia.nih.gov.

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


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