TY - JOUR
T1 - Human RAD52 interactions with replication protein a and the RAD51 presynaptic complex
AU - Ma, Chu Jian
AU - Kwon, Youngho
AU - Sung, Patrick
AU - Greene, Eric C.
N1 - Funding Information:
This work was supported by National Institutes of Health Grants R35GM118026 (to E. C. G.) and R01ES007061 and P01CA92584 (to P. S.). The authors declare that they have no conflicts of interest with the contents of this article. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
PY - 2017/7/14
Y1 - 2017/7/14
N2 - Rad52 is a highly conserved protein involved in the repair of DNA damage. Human RAD52 has been shown to mediate singlestranded DNA (ssDNA) and is synthetic lethal with mutations in other key recombination proteins. For this study, we used single-molecule imaging and ssDNA curtains to examine the binding interactions of human RAD52 with replication protein A (RPA)-coated ssDNA, and we monitored the fate of RAD52 during assembly of the presynaptic complex. We show that RAD52 binds tightly to the RPA-ssDNA complex and imparts an inhibitory effect on RPA turnover. We also found that during presynaptic complex assembly, most of the RPA and RAD52 was displaced from the ssDNA, but some RAD52-RPAssDNA complexes persisted as interspersed clusters surrounded by RAD51 filaments. Once assembled, the presence of RAD51 restricted formation of new RAD52-binding events, but additional RAD52 could bind once RAD51 dissociated from the ssDNA. Together, these results provide new insights into the behavior and dynamics of human RAD52 during presynaptic complex assembly and disassembly.
AB - Rad52 is a highly conserved protein involved in the repair of DNA damage. Human RAD52 has been shown to mediate singlestranded DNA (ssDNA) and is synthetic lethal with mutations in other key recombination proteins. For this study, we used single-molecule imaging and ssDNA curtains to examine the binding interactions of human RAD52 with replication protein A (RPA)-coated ssDNA, and we monitored the fate of RAD52 during assembly of the presynaptic complex. We show that RAD52 binds tightly to the RPA-ssDNA complex and imparts an inhibitory effect on RPA turnover. We also found that during presynaptic complex assembly, most of the RPA and RAD52 was displaced from the ssDNA, but some RAD52-RPAssDNA complexes persisted as interspersed clusters surrounded by RAD51 filaments. Once assembled, the presence of RAD51 restricted formation of new RAD52-binding events, but additional RAD52 could bind once RAD51 dissociated from the ssDNA. Together, these results provide new insights into the behavior and dynamics of human RAD52 during presynaptic complex assembly and disassembly.
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U2 - 10.1074/jbc.M117.794545
DO - 10.1074/jbc.M117.794545
M3 - Article
C2 - 28551686
AN - SCOPUS:85024362514
VL - 292
SP - 11702
EP - 11713
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 28
ER -