The Homeodomain Region of Rag-1 Reveals the Parallel Mechanisms of Bacterial and V(D)J Recombination
Abstract
The V(D)J recombinase subunits Rag-1 and Rag-2 mediate assembly of antigen receptor gene segments. We studied the mechanisms of DNA recognition by Rag-1/Rag-2 using surface plasmon resonance. The critical step for signal recognition is binding of Rag-1 to the nonamer. This is achieved by a region of Rag-1 homologous to the DNA-binding domain of the Hin family of bacterial invertases and to homeodomain proteins. Strikingly, the Hin homeodomain can functionally substitute for the Rag-1 homologous region. Rag-1 also interacts with the heptamer but with low affinity. Rag-2 shows no direct binding to DNA. Once the Rag-1/Rag-2 complex is engaged on the DNA, subsequent cleavage is directed by the heptamer sequence. This order of events remarkably parallels mechanisms that mediate transposition in bacteria and nematodes.
Additional Information
© 1996 Cell Press. Under an Elsevier user license. Received 18 June 1996, Revised 28 August 1996. Correspondence should be addressed to E. S. The authors would like to thank Dr. Patricia Cortes for valuable suggestions, Dr. Reid Johnson for the Hin clone, Drs. David Schatz and Christopher Roman for critical reading of the manuscript, and Drs. William Farley and Lesley Stolz of Pharmacia for the initial experiments with the BIAcore. E. S. is grateful to Christopher Roman, Patricia Cortes, and Dina Alexandropoulos for their generosity. G. P. would like to thank Mike Waterfield for his support. This work was supported by a Leukemia Research Foundation grant to E. S.Additional details
- Eprint ID
- 102739
- DOI
- 10.1016/s0092-8674(00)81344-6
- Resolver ID
- CaltechAUTHORS:20200422-151000513
- Leukemia Research Foundation
- Created
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2020-04-22Created from EPrint's datestamp field
- Updated
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2021-11-16Created from EPrint's last_modified field