The motor activity of DNA2 functions as an ssDNA translocase to promote DNA end resection

Genes Dev. 2017 Mar 1;31(5):493-502. doi: 10.1101/gad.295196.116. Epub 2017 Mar 23.

Abstract

DNA2 nuclease-helicase functions in DNA replication and recombination. This requires the nuclease of DNA2, while, in contrast, the role of the helicase activity has been unclear. We now show that the motor activity of both recombinant yeast and human DNA2 promotes efficient degradation of long stretches of ssDNA, particularly in the presence of the replication protein A. This degradation is further stimulated by a direct interaction with a cognate RecQ family helicase, which functions with DNA2 in DNA end resection to initiate homologous recombination. Consequently, helicase-deficient yeast dna2 K1080E cells display reduced resection speed of HO-induced DNA double-strand breaks. These results support a model of DNA2 and the RecQ family helicase partner forming a bidirectional motor machine, where the RecQ family helicase is the lead helicase, and the motor of DNA2 functions as a ssDNA translocase to promote degradation of 5'-terminated DNA.

Keywords: DNA end resection; DNA helicase; DNA nuclease; hDNA2; ssDNA translocase; yDna2.

MeSH terms

  • DNA End-Joining Repair / genetics
  • DNA End-Joining Repair / physiology*
  • DNA Helicases / metabolism*
  • DNA, Single-Stranded / metabolism*
  • Homologous Recombination
  • Humans
  • RecQ Helicases / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism*
  • Replication Protein A / metabolism
  • Saccharomyces cerevisiae / enzymology*
  • Saccharomyces cerevisiae / genetics

Substances

  • DNA, Single-Stranded
  • Recombinant Proteins
  • Replication Protein A
  • DNA Helicases
  • RecQ Helicases