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Molecular and Cellular Biology, September 2005, p. 8074-8083, Vol. 25, No. 18
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.18.8074-8083.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Role of the Schizosaccharomyces pombe F-Box DNA Helicase in Processing Recombination Intermediates

Takashi Morishita,1 Fumiko Furukawa,1 Chikako Sakaguchi,1 Takashi Toda,2 Antony M. Carr,3 Hiroshi Iwasaki,4 and Hideo Shinagawa1*

Department of Molecular Microbiology, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamada-oka, Suita, Osaka 565-0871, Japan,1 Laboratory of Cell Regulation, Cancer Research UK, London Research Institute, Lincoln's Inn Fields Laboratories, 44 Lincoln's Inn Fields, London WC2A 3PX, United Kingdom,2 Genome Damage and Stability Centre, University of Sussex, Brighton, BN1 9RQ, United Kingdom,3 Graduate School of Integrated Science, Yokohama City University, 1-7-29 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan4

Received 3 March 2005/ Returned for modification 10 April 2005/ Accepted 23 June 2005

In an effort to identify novel genes involved in recombination repair, we isolated fission yeast Schizosaccharomyces pombe mutants sensitive to methyl methanesulfonate (MMS) and a synthetic lethal with rad2. A gene that complements such mutations was isolated from the S. pombe genomic library, and subsequent analysis identified it as the fbh1 gene encoding the F-box DNA helicase, which is conserved in mammals but not conserved in Saccharomyces cerevisiae. An fbh1 deletion mutant is moderately sensitive to UV, MMS, and {gamma} rays. The rhp51 (RAD51 ortholog) mutation is epistatic to fbh1. fbh1 is essential for viability in stationary-phase cells and in the absence of either Srs2 or Rqh1 DNA helicase. In each case, lethality is suppressed by deletion of the recombination gene rhp57. These results suggested that fbh1 acts downstream of rhp51 and rhp57. Following UV irradiation or entry into the stationary phase, nuclear chromosomal domains of the fbh1{Delta} mutant shrank, and accumulation of some recombination intermediates was suggested by pulsed-field gel electrophoresis. Focus formation of Fbh1 protein was induced by treatment that damages DNA. Thus, the F-box DNA helicase appears to process toxic recombination intermediates, the formation of which is dependent on the function of Rhp51.

* Corresponding author. Mailing address: Department of Molecular Microbiology, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamada-oka, Suita, Osaka 565-0871, Japan. Phone: 81-6-6879-8317. Fax: 81-6-6879-8320. E-mail: shinagaw{at}biken.osaka-u.ac.jp.

Molecular and Cellular Biology, September 2005, p. 8074-8083, Vol. 25, No. 18
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.18.8074-8083.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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