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J. Biol. Chem., Vol. 280, Issue 46, 38631-38638, November 18, 2005

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The Saccharomyces cerevisiae Sae2 Protein Promotes Resection and Bridging of Double Strand Break Ends^*

From the Dipartimento di Biotecnologie e Bioscienze, Università di Milano-Bicocca, P.zza della Scienza 2, 20126 Milano, Italy

When eukaryotic chromosomes undergo double strand breaks (DSBs), several evolutionarily conserved proteins, among which the MRX complex, are recruited to the break site, leading to checkpoint activation and DNA repair. The function of the Saccharomyces cerevisiae Sae2 protein, which is known to work together with the MRX complex in meiotic DSB processing and in specific mitotic DSB repair events, is only beginning to be elucidated. Here we provide new insights into the role of Sae2 in mitotic DSB repair. We show that repair by single strand annealing of a single DSB, which is generated by the HO endonuclease between direct repeats, is defective both in the absence of Sae2 and in the presence of the hypomorphic rad50s allele altering the Rad50 subunit of MRX. Moreover, SAE2 overexpression partially suppresses the rad50s single strand annealing repair defects, suggesting that the latter might arise from defective MRX-Sae2 interactions. Finally, SAE2 deletion slows down resection of an HO-induced DSB and impairs DSB end bridging. Thus, Sae2 participates in DSB single strand annealing repair by ensuring both resection and intrachromosomal association of the broken ends.

Received for publication, July 29, 2005 , and in revised form, September 7, 2005.

^* This work was supported by grants from Associazione Italiana per la Ricerca sul Cancro, Cofinanziamento 2003 MIUR/Università di Milano-Bicocca and European Community's Human Potential Programme HPRN-CT-2002-00238 (to M. P. L.), and from Fondo per gli Investimenti della Ricerca di Base (FIRB) (to G. L.). 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.

¹ Both authors contributed equally to this work.

² To whom correspondence should be addressed. Tel.: 0039-0264483425; Fax: 0039-0264483565; E-mail: mariapia.longhese{at}unimib.it.

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