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Interplay of Mre11 Nuclease with Dna2 plus Sgs1 in Rad51-Dependent Recombinational Repair

Budd, Martin E. and Campbell, Judith L. (2009) Interplay of Mre11 Nuclease with Dna2 plus Sgs1 in Rad51-Dependent Recombinational Repair. PLoS ONE, 4 (1). Art. No. e4267. ISSN 1932-6203. PMCID PMC2625443. https://resolver.caltech.edu/CaltechAUTHORS:20090417-105037556

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Abstract

The Mre11/Rad50/Xrs2 complex initiates IR repair by binding to the end of a double-strand break, resulting in 5′ to 3′ exonuclease degradation creating a single-stranded 3′ overhang competent for strand invasion into the unbroken chromosome. The nuclease(s) involved are not well understood. Mre11 encodes a nuclease, but it has 3′ to 5′, rather than 5′ to 3′ activity. Furthermore, mutations that inactivate only the nuclease activity of Mre11 but not its other repair functions, mre11-D56N and mre11-H125N, are resistant to IR. This suggests that another nuclease can catalyze 5′ to 3′ degradation. One candidate nuclease that has not been tested to date because it is encoded by an essential gene is the Dna2 helicase/nuclease. We recently reported the ability to suppress the lethality of a dna2Δ with a pif1Δ. The dna2Δ pif1Δ mutant is IR-resistant. We have determined that dna2Δ pif1Δ mre11-D56N and dna2Δ pif1Δ mre11-H125N strains are equally as sensitive to IR as mre11Δ strains, suggesting that in the absence of Dna2, Mre11 nuclease carries out repair. The dna2Δ pif1Δ mre11-D56N triple mutant is complemented by plasmids expressing Mre11, Dna2 or dna2K1080E, a mutant with defective helicase and functional nuclease, demonstrating that the nuclease of Dna2 compensates for the absence of Mre11 nuclease in IR repair, presumably in 5′ to 3′ degradation at DSB ends. We further show that sgs1Δ mre11-H125N, but not sgs1Δ, is very sensitive to IR, implicating the Sgs1 helicase in the Dna2-mediated pathway.


Item Type:Article
Related URLs:
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http://dx.doi.org/10.1371/journal.pone.0004267DOIArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2625443/PubMed CentralArticle
Additional Information:© 2009 Campbell et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Received December 11, 2008; Accepted December 22, 2008; Published January 23, 2009. This work was supported by NIH GM087666, the Philip Morris External Research Program, and the Ellison Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We thank Kevin Lewis for the cloned wild-type and mutant MRE11 genes.
Funders:
Funding AgencyGrant Number
NIHGM087666
Philip Morris External Research ProgramUNSPECIFIED
Ellison Medical FoundationUNSPECIFIED
Issue or Number:1
PubMed Central ID:PMC2625443
Record Number:CaltechAUTHORS:20090417-105037556
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20090417-105037556
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:14008
Collection:CaltechAUTHORS
Deposited By: Jason Perez
Deposited On:21 Apr 2009 22:54
Last Modified:12 Feb 2020 21:29

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