Rad52-mediated DNA annealing after Rad51-mediated DNA strand exchange promotes second ssDNA capture

Tomohiko Sugiyama, Noriko Kantake, Yun Wu, Stephen C. Kowalczykowski

Research output: Contribution to journalArticle

79 Citations (Scopus)

Abstract

Rad51, Rad52, and RPA play central roles in homologous DNA recombination. Rad51 mediates DNA strand exchange, a key reaction in DNA recombination. Rad52 has two distinct activities: to recruit Rad51 onto single-strand (ss)DNA that is complexed with the ssDNA-binding protein, RPA, and to anneal complementary ssDNA complexed with RPA. Here, we report that Rad52 promotes annealing of the ssDNA strand that is displaced by DNA strand exchange by Rad51 and RPA, to a second ssDNA strand. An RPA that is recombination-deficient (RPA(rfa1-t11)) failed to support annealing, explaining its in vivo phenotype. Escherichia coli RecO and SSB proteins, which are functional homologues of Rad52 and RPA, also facilitated the same reaction, demonstrating its conserved nature. We also demonstrate that the two activities of Rad52, recruiting Rad51 and annealing DNA, are coordinated in DNA strand exchange and second ssDNA capture.

Original languageEnglish (US)
Pages (from-to)5539-5548
Number of pages10
JournalEMBO Journal
Volume25
Issue number23
DOIs
StatePublished - Nov 29 2006

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Annealing
DNA
Genetic Recombination
Homologous Recombination
Escherichia coli
Carrier Proteins
Phenotype
Proteins

Keywords

  • DNA annealing
  • DNA repair
  • Rad52
  • Recombination
  • Second ssDNA capture

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

Rad52-mediated DNA annealing after Rad51-mediated DNA strand exchange promotes second ssDNA capture. / Sugiyama, Tomohiko; Kantake, Noriko; Wu, Yun; Kowalczykowski, Stephen C.

In: EMBO Journal, Vol. 25, No. 23, 29.11.2006, p. 5539-5548.

Research output: Contribution to journalArticle

Sugiyama, Tomohiko ; Kantake, Noriko ; Wu, Yun ; Kowalczykowski, Stephen C. / Rad52-mediated DNA annealing after Rad51-mediated DNA strand exchange promotes second ssDNA capture. In: EMBO Journal. 2006 ; Vol. 25, No. 23. pp. 5539-5548.
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AB - Rad51, Rad52, and RPA play central roles in homologous DNA recombination. Rad51 mediates DNA strand exchange, a key reaction in DNA recombination. Rad52 has two distinct activities: to recruit Rad51 onto single-strand (ss)DNA that is complexed with the ssDNA-binding protein, RPA, and to anneal complementary ssDNA complexed with RPA. Here, we report that Rad52 promotes annealing of the ssDNA strand that is displaced by DNA strand exchange by Rad51 and RPA, to a second ssDNA strand. An RPA that is recombination-deficient (RPA(rfa1-t11)) failed to support annealing, explaining its in vivo phenotype. Escherichia coli RecO and SSB proteins, which are functional homologues of Rad52 and RPA, also facilitated the same reaction, demonstrating its conserved nature. We also demonstrate that the two activities of Rad52, recruiting Rad51 and annealing DNA, are coordinated in DNA strand exchange and second ssDNA capture.

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