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Molecular and Cellular Biology, February 2005, p. 933-944, Vol. 25, No. 3
0270-7306/05/$08.00+0 doi:10.1128/MCB.25.3.933-944.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.
RAD51-Dependent Break-Induced Replication Differs in Kinetics and Checkpoint Responses from RAD51-Mediated Gene Conversion
Anna Malkova ,
,
Maria L. Naylor,
,
Miyuki Yamaguchi,
Grzegorz Ira, and
James E. Haber*
Department of Biology and Rosenstiel Center, Brandeis University, Waltham, Massachusetts
Received 13 August 2004/
Returned for modification 22 September 2004/
Accepted 25 October 2004
Diploid Saccharomyces cells experiencing a double-strand break (DSB) on one homologous chromosome repair the break by RAD51-mediated gene conversion >98% of the time. However, when extensive homologous sequences are restricted to one side of the DSB, repair can occur by both RAD51-dependent and RAD51-independent break-induced replication (BIR) mechanisms. Here we characterize the kinetics and checkpoint dependence of RAD51-dependent BIR when the DSB is created within a chromosome. Gene conversion products appear within 2 h, and there is little, if any, induction of the DNA damage checkpoint; however, RAD51-dependent BIR occurs with a further delay of 2 to 4 h and cells arrest in response to the G2/M DNA damage checkpoint. RAD51-dependent BIR does not require special facilitating sequences that are required for a less efficient RAD51-independent process. RAD51-dependent BIR occurs efficiently in G2-arrested cells. Once repair is initiated, the rate of repair replication during BIR is comparable to that of normal DNA replication, as copying of >100 kb is completed less than 30 min after repair DNA synthesis is detected close to the DSB.
* Corresponding author. Mailing address: Rosenstiel Center, Brandeis University, 415 South St., Mail Stop 029, Waltham, MA 02454-9110. Phone: (781) 736-2462. Fax: (781) 736-2405. E-mail:
haber{at}brandeis.edu.
A.M., M.L.N., and M.Y. contributed equally to this work.
Present address: Biology Department, Indiana UniversityPurdue University Indianapolis, Indianapolis, IN 46202.
Present address: Department of Genetics, Harvard Medical School, Boston, MA 02115.
Molecular and Cellular Biology, February 2005, p. 933-944, Vol. 25, No. 3
0022-538X/05/$08.00+0 doi:10.1128/MCB.25.3.933-944.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.
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