Joining Mutants of RAG1 and RAG2 that Demonstrate Impaired Interactions with the Coding-end DNA*
- Department of Biophysics and Biochemistry, Graduate School of Science, and Core Research for Evolutional Science and Technology Program of the Japan Science and Technology Agency, The University of Tokyo, Bunkyo-ku, Tokyo 113-0032, Japan
- § To whom correspondence should be addressed. Tel.: 81-3-5841-7239; Fax: 81-3-5841-7240; E-mail: sakano{at}mail.ecc.u-tokyo.ac.jp.
Abstract
In V(D)J joining of antigen receptor genes, two recombination signal sequences (RSSs), 12- and 23-RSSs, form a complex with the protein products of recombination activating genes, RAG1 and RAG2. DNaseI footprinting demonstrates that the interaction of RAG proteins with substrate RSS DNA is not just limited to the signal region but involves the coding sequence as well. Joining mutants of RAG1 and RAG2 demonstrate impaired interactions with the coding region in both pre- and postcleavage type complexes. A possible role of this RAG coding region interaction is discussed in the context of V(D)J recombination.
Footnotes
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↵1 The abbreviations used are: RSS, recombination signal sequence; SE, signal end; CE, coding end; MOPS, 4-morpholinepropanesulfonic acid; dsDNA, double strand DNA.
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↵2 F. Nagawa, S. Hirose, H. Nishizumi, T. Nishihara, and H. Sakano, unpublished observation.
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↵* This work was supported in part by grants from the Ministry of Education Culture and Science of Japan and the Japan Science and Technology Agency. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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↵‡ Both authors contributed equally to this work.
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- Received May 17, 2004.
- Revision received June 16, 2004.
- The American Society for Biochemistry and Molecular Biology, Inc.
