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Molecular and Cellular Biology, November 1998, p. 6430-6435, Vol. 18, No. 11
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Homologous Recombination, but Not DNA Repair, Is Reduced in Vertebrate Cells Deficient in RAD52

Yuko Yamaguchi-Iwai,1 Eiichiro Sonoda,1 Jean-Marie Buerstedde,2,dagger Olga Bezzubova,2,dagger Ciaran Morrison,1 Minoru Takata,1 Akira Shinohara,3,Dagger and Shunichi Takeda1,*

Bayer-chair Department of Molecular Immunology and Allergology, Faculty of Medicine, Kyoto University, Konoe Yoshida, Sakyo-ku, Kyoto 606-8501,1 and Department of Biology, Faculty of Science, Osaka University, Toyonaka, Osaka 560-0043,3 Japan, and Basel Institute for Immunology, CH-4005 Basel, Switzerland2

Received 30 April 1998/Returned for modification 1 June 1998/Accepted 27 July 1998

Rad52 plays a pivotal role in double-strand break (DSB) repair and genetic recombination in Saccharomyces cerevisiae, where mutation of this gene leads to extreme X-ray sensitivity and defective recombination. Yeast Rad51 and Rad52 interact, as do their human homologues, which stimulates Rad51-mediated DNA strand exchange in vitro, suggesting that Rad51 and Rad52 act cooperatively. To define the role of Rad52 in vertebrates, we generated RAD52-/- mutants of the chicken B-cell line DT40. Surprisingly, RAD52-/- cells were not hypersensitive to DNA damages induced by gamma -irradiation, methyl methanesulfonate, or cis-platinum(II)diammine dichloride (cisplatin). Intrachromosomal recombination, measured by immunoglobulin gene conversion, and radiation-induced Rad51 nuclear focus formation, which is a putative intermediate step during recombinational repair, occurred as frequently in RAD52-/- cells as in wild-type cells. Targeted integration frequencies, however, were consistently reduced in RAD52-/- cells, showing a clear role for Rad52 in genetic recombination. These findings reveal striking differences between S. cerevisiae and vertebrates in the functions of RAD51 and RAD52.

* Corresponding author. Mailing address: Bayer-chair Department of Molecular Immunology and Allergology, Faculty of Medicine, Kyoto University, Konoe Yoshida, Sakyo-ku, Kyoto 606-8501, Japan. Phone: 81-75-771-8159. Fax: 81-75-771-8184. E-mail: stakeda{at}mfour.med.kyoto-u.ac.jp.

dagger Present address: Department of Cellular Immunology, Heinrich-Pette-Institute, 20251 Hamburg, Germany.

Dagger Present address: Department of Radiation and Cellular Oncology, University of Chicago, Chicago, IL 60637.

Molecular and Cellular Biology, November 1998, p. 6430-6435, Vol. 18, No. 11
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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