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 language | English (US) |
|---|---|
| Pages (from-to) | 5539-5548 |
| Number of pages | 10 |
| Journal | EMBO Journal |
| Volume | 25 |
| Issue number | 23 |
| DOIs | |
| State | Published - Nov 29 2006 |
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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 journal › Article
}
TY - JOUR
T1 - Rad52-mediated DNA annealing after Rad51-mediated DNA strand exchange promotes second ssDNA capture
AU - Sugiyama, Tomohiko
AU - Kantake, Noriko
AU - Wu, Yun
AU - Kowalczykowski, Stephen C.
PY - 2006/11/29
Y1 - 2006/11/29
N2 - 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.
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.
KW - DNA annealing
KW - DNA repair
KW - Rad52
KW - Recombination
KW - Second ssDNA capture
UR - http://www.scopus.com/inward/record.url?scp=33751560821&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33751560821&partnerID=8YFLogxK
U2 - 10.1038/sj.emboj.7601412
DO - 10.1038/sj.emboj.7601412
M3 - Article
C2 - 17093500
AN - SCOPUS:33751560821
VL - 25
SP - 5539
EP - 5548
JO - EMBO Journal
JF - EMBO Journal
SN - 0261-4189
IS - 23
ER -