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Nucleic Acids Res November 1, 2008; 36 (19): 6091-100.

Identification of the Xenopus DNA2 protein as a major nuclease for the 5''->3'' strand-specific processing of DNA ends.

Liao S , Toczylowski T , Yan H .

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 5''-->3'' 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 5''-->3'' 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.

PubMed ID: 18820296
PMC ID: PMC2577336
Article link: Nucleic Acids Res
Grant support: [+]

Species referenced: Xenopus
Genes referenced: dna2 mre11 rbbp8 wrn

Article Images: [+] show captions
References [+] :
Amundsen, Interchangeable parts of the Escherichia coli recombination machinery. 2003, Pubmed