TY - JOUR
T1 - Novel function of HATs and HDACs in homologous recombination through acetylation of human RAD52 at double-strand break sites
AU - Yasuda, Takeshi
AU - Kagawa, Wataru
AU - Ogi, Tomoo
AU - Kato, Takamitsu A.
AU - Suzuki, Takehiro
AU - Dohmae, Naoshi
AU - Takizawa, Kazuya
AU - Nakazawa, Yuka
AU - Genet, Matthew D.
AU - Saotome, Mika
AU - Hama, Michio
AU - Konishi, Teruaki
AU - Nakajima, Nakako Izumi
AU - Hazawa, Masaharu
AU - Tomita, Masanori
AU - Koike, Manabu
AU - Noshiro, Katsuko
AU - Tomiyama, Kenichi
AU - Obara, Chizuka
AU - Gotoh, Takaya
AU - Ui, Ayako
AU - Fujimori, Akira
AU - Nakayama, Fumiaki
AU - Hanaoka, Fumio
AU - Sugasawa, Kaoru
AU - Okayasu, Ryuichi
AU - Jeggo, Penny A.
AU - Tajima, Katsushi
N1 - Funding Information:
This study was funded by Grants-in Aid from the Ministry of Education, Culture, Sports, Science, and Technology of Japan 26281026 to TY, the Futaba Electronics Memorial Foundation H20-042 to TY, Collaborative Research and Shared Equipment of Atomic Bomb Disease Institute, Nagasaki University A-2-4 and A-2-23 to TY, the Program of the network-type joint Usage/Research Center for Radiation Disaster Medical Science of Hiroshima University, Nagasaki University, and Fukushima Medical University to TY, the joint research program of Biosignal Research Center, Kobe University 281004 to TY, and the general fund of the NIRS IOL to RO. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We thank Dr. D. Livingston (Dana-Farber Cancer Institute and Harvard Medical School, Boston, MA, USA), Dr. R. Goodman (Oregon Health Sciences University, Portland, OR, USA), and Dr. K. Yokoyama (RIKEN, Tsukuba, Japan) for the p300/CBP expression constructs; Dr. M. Jasin (Memorial Sloan-Kettering Cancer Center, NYC, USA) and Dr. J.M. Stark (Beckman Research Institute of City of Hope, CA, USA) for materials for the DR-GFP assay; Dr. K. Komaki-Yasuda (NCGM, Tokyo, Japan) for the protein sequence analysis; Dr. A. Saotome-Nakamura (NIRS, Chiba, Japan), K. Matsumoto (Meisei University, Tokyo) and Y. Aizawa (Meisei University, Tokyo) for helpful support of experiments; and Dr. A. Yasui (Tohoku University, Sendai, Japan) for helpful comments. We deeply appreciate Dr. H. Kurumizaka and his laboratory members (Waseda University, Tokyo, Japan) for materials and helpful advice.
Publisher Copyright:
© 2018 Yasuda et al.
PY - 2018/3
Y1 - 2018/3
N2 - The p300 and CBP histone acetyltransferases are recruited to DNA double-strand break (DSB) sites where they induce histone acetylation, thereby influencing the chromatin structure and DNA repair process. Whether p300/CBP at DSB sites also acetylate non-histone proteins, and how their acetylation affects DSB repair, remain unknown. Here we show that p300/CBP acetylate RAD52, a human homologous recombination (HR) DNA repair protein, at DSB sites. Using in vitro acetylated RAD52, we identified 13 potential acetylation sites in RAD52 by a mass spectrometry analysis. An immunofluorescence microscopy analysis revealed that RAD52 acetylation at DSBs sites is counteracted by SIRT2- and SIRT3-mediated deacetylation, and that non-acetylated RAD52 initially accumulates at DSB sites, but dissociates prematurely from them. In the absence of RAD52 acetylation, RAD51, which plays a central role in HR, also dissociates prematurely from DSB sites, and hence HR is impaired. Furthermore, inhibition of ataxia telangiectasia mutated (ATM) protein by siRNA or inhibitor treatment demonstrated that the acetylation of RAD52 at DSB sites is dependent on the ATM protein kinase activity, through the formation of RAD52, p300/CBP, SIRT2, and SIRT3 foci at DSB sites. Our findings clarify the importance of RAD52 acetylation in HR and its underlying mechanism.
AB - The p300 and CBP histone acetyltransferases are recruited to DNA double-strand break (DSB) sites where they induce histone acetylation, thereby influencing the chromatin structure and DNA repair process. Whether p300/CBP at DSB sites also acetylate non-histone proteins, and how their acetylation affects DSB repair, remain unknown. Here we show that p300/CBP acetylate RAD52, a human homologous recombination (HR) DNA repair protein, at DSB sites. Using in vitro acetylated RAD52, we identified 13 potential acetylation sites in RAD52 by a mass spectrometry analysis. An immunofluorescence microscopy analysis revealed that RAD52 acetylation at DSBs sites is counteracted by SIRT2- and SIRT3-mediated deacetylation, and that non-acetylated RAD52 initially accumulates at DSB sites, but dissociates prematurely from them. In the absence of RAD52 acetylation, RAD51, which plays a central role in HR, also dissociates prematurely from DSB sites, and hence HR is impaired. Furthermore, inhibition of ataxia telangiectasia mutated (ATM) protein by siRNA or inhibitor treatment demonstrated that the acetylation of RAD52 at DSB sites is dependent on the ATM protein kinase activity, through the formation of RAD52, p300/CBP, SIRT2, and SIRT3 foci at DSB sites. Our findings clarify the importance of RAD52 acetylation in HR and its underlying mechanism.
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U2 - 10.1371/journal.pgen.1007277
DO - 10.1371/journal.pgen.1007277
M3 - Article
C2 - 29590107
AN - SCOPUS:85044828012
VL - 14
JO - PLoS Genetics
JF - PLoS Genetics
SN - 1553-7390
IS - 3
M1 - e1007277
ER -