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Molecular and Cellular Biology, March 2006, p. 1617-1630, Vol. 26, No. 5
0270-7306/06/$08.00+0     doi:10.1128/MCB.26.5.1617-1630.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

The Nse5-Nse6 Dimer Mediates DNA Repair Roles of the Smc5-Smc6 Complex{dagger}

Stephanie Pebernard,1 James Wohlschlegel,2 W. Hayes McDonald,2 John R. Yates III,2 and Michael N. Boddy1*

Department of Molecular Biology,1 Department of Cell Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 920372

Received 26 September 2005/ Returned for modification 19 October 2005/ Accepted 6 December 2005

Stabilization and processing of stalled replication forks is critical for cell survival and genomic integrity. We characterize a novel DNA repair heterodimer of Nse5 and Nse6, which are nonessential nuclear proteins critical for chromosome segregation in fission yeast. The Nse5/6 dimer facilitates DNA repair as part of the Smc5-Smc6 holocomplex (Smc5/6), the basic architecture of which we define. Nse5-Nes6 (Nse5/6) mutants display a high level of spontaneous DNA damage and mitotic catastrophe in the absence of the master checkpoint regulator Rad3 (hATR). Nse5/6 mutants are required for the response to genotoxic agents that block the progression of replication forks, acting in a pathway that allows the tolerance of irreparable UV lesions. Interestingly, the UV sensitivity of Nse5/6 mutants is suppressed by concomitant deletion of the homologous recombination repair factor, Rhp51 (Rad51). Further, the viability of Nse5/6 mutants depends on Mus81 and Rqh1, factors that resolve or prevent the formation of Holliday junctions. Consistently, the UV sensitivity of cells lacking Nse5/6 can be partially suppressed by overexpressing the bacterial resolvase RusA. We propose a role for Nse5/6 mutants in suppressing recombination that results in Holliday junction formation or in Holliday junction resolution.

* Corresponding author. Mailing address: The Scripps Research Institute, Rm. MB107, 10550 North Torrey Pines Rd., Molecular Biology, MB-3, La Jolla, CA 92037. Phone: (858) 784-7042. Fax: (858) 784-2265. E-mail: nboddy{at}scripps.edu.

{dagger} M.N.B. dedicates this study to his son Conor.

Molecular and Cellular Biology, March 2006, p. 1617-1630, Vol. 26, No. 5
0022-538X/06/$08.00+0     doi:10.1128/MCB.26.5.1617-1630.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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