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Originally published In Press as doi:10.1074/jbc.C200337200 on August 21, 2002

J. Biol. Chem., Vol. 277, Issue 42, 39679-39683, October 18, 2002
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RH70, a Bidirectional RNA Helicase, Co-purifies with U1snRNP*

Chee-Gun LeeDagger

From the Department of Biochemistry and Molecular Biology, Graduate School of Biomedical Sciences, University of Medicine and Dentistry of New Jersey, Newark, New Jersey 07103

In this report, I describe the co-purification of a novel 70-kDa RNA helicase (RH70) and U1snRNP through six column steps. Peptide sequence analysis by mass spectrometry and Edman degradation revealed that RH70 is the previously reported DDX17. Biochemical characterization of RH70, obtained by partial separation from U1snRNP, yielded the following results. (a) RH70 mediates the unwinding of duplex RNA but not DNA in an ATP-dependent manner. (b) Both the RNA-dependent ATPase and RNA helicase activities of RH70 are highly specific for ATP, exhibiting an apparent Km of 0.5 mM. (c) RH70 catalyzes the unwinding of duplex RNA containing single-stranded regions at either the 5'- or 3'-end. Its association with U1snRNP and ATP specificity suggest a role for RH70 in pre-mRNA splicing, in particular, at the early stages of the splicing reaction involving U1snRNP.

* This study was supported by a research grant from the University of Medicine and Dentistry of New Jersey (UMDNJ) Foundation and the UMDNJ-Dean's biomedical research support grant program.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 To whom correspondence should be addressed: Dept. of Biochemistry and Molecular Biology, Graduate School of Biomedical Sciences, University of Medicine and Dentistry of New Jersey, 185 South Orange Ave., Newark, NJ 07103. Tel.: 973-972-0130; Fax: 973-972-5594; E-mail: leecg@umdnj.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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