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Roles of C-Terminal Region of Yeast and Human Rad52 in Rad51-Nucleoprotein Filament Formation and ssDNA Annealing.
PLoS One. 2016; 11(6):e0158436.Plos

Abstract

Yeast Rad52 (yRad52) has two important functions at homologous DNA recombination (HR); annealing complementary single-strand DNA (ssDNA) molecules and recruiting Rad51 recombinase onto ssDNA (recombination mediator activity). Its human homolog (hRAD52) has a lesser role in HR, and apparently lacks mediator activity. Here we show that yRad52 can load human Rad51 (hRAD51) onto ssDNA complexed with yeast RPA in vitro. This is biochemically equivalent to mediator activity because it depends on the C-terminal Rad51-binding region of yRad52 and on functional Rad52-RPA interaction. It has been reported that the N-terminal two thirds of both yRad52 and hRAD52 is essential for binding to and annealing ssDNA. Although a second DNA binding region has been found in the C-terminal region of yRad52, its role in ssDNA annealing is not clear. In this paper, we also show that the C-terminal region of yRad52, but not of hRAD52, is involved in ssDNA annealing. This suggests that the second DNA binding site is required for the efficient ssDNA annealing by yRad52. We propose an updated model of Rad52-mediated ssDNA annealing.

Authors+Show Affiliations

Department of Biological Sciences, Ohio University, Athens, Ohio, United States of America.Department of Biological Sciences, Ohio University, Athens, Ohio, United States of America.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

27362509

Citation

Khade, Nilesh V., and Tomohiko Sugiyama. "Roles of C-Terminal Region of Yeast and Human Rad52 in Rad51-Nucleoprotein Filament Formation and ssDNA Annealing." PloS One, vol. 11, no. 6, 2016, pp. e0158436.
Khade NV, Sugiyama T. Roles of C-Terminal Region of Yeast and Human Rad52 in Rad51-Nucleoprotein Filament Formation and ssDNA Annealing. PLoS One. 2016;11(6):e0158436.
Khade, N. V., & Sugiyama, T. (2016). Roles of C-Terminal Region of Yeast and Human Rad52 in Rad51-Nucleoprotein Filament Formation and ssDNA Annealing. PloS One, 11(6), e0158436. https://doi.org/10.1371/journal.pone.0158436
Khade NV, Sugiyama T. Roles of C-Terminal Region of Yeast and Human Rad52 in Rad51-Nucleoprotein Filament Formation and ssDNA Annealing. PLoS One. 2016;11(6):e0158436. PubMed PMID: 27362509.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Roles of C-Terminal Region of Yeast and Human Rad52 in Rad51-Nucleoprotein Filament Formation and ssDNA Annealing. AU - Khade,Nilesh V, AU - Sugiyama,Tomohiko, Y1 - 2016/06/30/ PY - 2016/03/17/received PY - 2016/06/15/accepted PY - 2016/7/1/entrez PY - 2016/7/1/pubmed PY - 2017/7/19/medline SP - e0158436 EP - e0158436 JF - PloS one JO - PLoS One VL - 11 IS - 6 N2 - Yeast Rad52 (yRad52) has two important functions at homologous DNA recombination (HR); annealing complementary single-strand DNA (ssDNA) molecules and recruiting Rad51 recombinase onto ssDNA (recombination mediator activity). Its human homolog (hRAD52) has a lesser role in HR, and apparently lacks mediator activity. Here we show that yRad52 can load human Rad51 (hRAD51) onto ssDNA complexed with yeast RPA in vitro. This is biochemically equivalent to mediator activity because it depends on the C-terminal Rad51-binding region of yRad52 and on functional Rad52-RPA interaction. It has been reported that the N-terminal two thirds of both yRad52 and hRAD52 is essential for binding to and annealing ssDNA. Although a second DNA binding region has been found in the C-terminal region of yRad52, its role in ssDNA annealing is not clear. In this paper, we also show that the C-terminal region of yRad52, but not of hRAD52, is involved in ssDNA annealing. This suggests that the second DNA binding site is required for the efficient ssDNA annealing by yRad52. We propose an updated model of Rad52-mediated ssDNA annealing. SN - 1932-6203 UR - https://www.unboundmedicine.com/medline/citation/27362509/Roles_of_C_Terminal_Region_of_Yeast_and_Human_Rad52_in_Rad51_Nucleoprotein_Filament_Formation_and_ssDNA_Annealing_ L2 - https://dx.plos.org/10.1371/journal.pone.0158436 DB - PRIME DP - Unbound Medicine ER -