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Originally published In Press as doi:10.1074/jbc.M103611200 on June 26, 2001

J. Biol. Chem., Vol. 276, Issue 37, 35024-35028, September 14, 2001
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A Unique beta -Hairpin Protruding from AAA+ ATPase Domain of RuvB Motor Protein Is Involved in the Interaction with RuvA DNA Recognition Protein for Branch Migration of Holliday Junctions*

Yong-Woon HanDagger , Hiroshi IwasakiDagger §, Tomoko Miyata**, Kouta Mayanagi**, Kazuhiro YamadaDagger **, Kosuke Morikawa**, and Hideo ShinagawaDagger ||

From the Dagger  Research Institute for Microbial Diseases, Osaka University 3-1 Yamadaoka, Suita, Osaka 565-0871, the § Japan Science and Technology Corporation Precursory Research for Embryonic Science and Technology, 3-1 Yamadaoka, Suita, Osaka 565-0871, and the ** Biomolecular Engineering Research Institute, 6-2-3 Furuedai, Suita, Osaka 565-0874, Japan

The Escherichia coli RuvB protein is a motor protein that forms a complex with RuvA and promotes branch migration of Holliday junctions during homologous recombination. This study describes the characteristics of two RuvB mutants, I148T and I150T, that do not promote branch migration in the presence of RuvA. These RuvB mutants hydrolyzed ATP and bound duplex DNA with the same efficiency as wild-type RuvB, but the mutants did not form a complex with RuvA and were defective in loading onto junction DNA in a RuvA-assisted manner. A recent crystallographic study revealed that Ile148 and Ile150 are in a unique beta -hairpin that protrudes from the AAA+ ATPase domain of RuvB. We propose that this beta -hairpin interacts with hydrophobic residues in the mobile third domain of RuvA and that this interaction is vital for the RuvA-assisted loading of RuvB onto Holliday junction DNA.

* This work was supported by Grants-in-aid for Scientific Research 08280102 and 0828010 on Priority Areas from the Ministry of Education, Science, Sports and Culture of Japan (to H. S.).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 may be addressed: Graduate School of Integrated Science, Yokohama City University, 1-7-29 Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan. Tel.: 81-45-508-7238; Fax: 81-45-508-7369; E-mail: iwasaki@tsurumi.yokohama-cu.ac.jp.

|| To whom correspondence may be addressed. Tel.: 81-6-6879-8317; Fax: 81-6-6879-8320; E-mail: shinagaw@biken.osaka-u.ac.jp.

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


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