Journal
NUCLEIC ACIDS RESEARCH
Volume 36, Issue 19, Pages 6091-6100Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkn616
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Funding
- National Institute of Health [R01 GM57962-02]
- Pennsylvania Department of Health [69133-01]
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The first step of homology-dependent DNA double-strand break (DSB) repair is the 5 strand-specific processing of DNA ends to generate 3 single-strand tails. Despite extensive effort, the nuclease(s) that is directly responsible for the resection of 5 strands in eukaryotic cells remains elusive. Using nucleoplasmic extracts (NPE) derived from the eggs of Xenopus laevis as the model system, we have found that DNA processing consists of at least two steps: an ATP-dependent unwinding of ends and an ATP-independent 53 degradation of single-strand tails. The unwinding step is catalyzed by DNA helicases, the major one of which is the Xenopus Werner syndrome protein (xWRN), a member of the RecQ helicase family. In this study, we report the purification and identification of the Xenopus DNA2 (xDNA2) as one of the nucleases responsible for the 53 degradation of single-strand tails. Immunodepletion of xDNA2 resulted in a significant reduction in end processing and homology-dependent DSB repair. These results provide strong evidence that xDNA2 is a major nuclease for the resection of DNA ends for homology-dependent DSB repair in eukaryotes.
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