TY - JOUR
T1 - RAP80 suppresses the vulnerability of R-loops during DNA double-strand break repair
AU - Yasuhara, Takaaki
AU - Kato, Reona
AU - Yamauchi, Motohiro
AU - Uchihara, Yuki
AU - Zou, Lee
AU - Miyagawa, Kiyoshi
AU - Shibata, Atsushi
N1 - Funding Information:
We thank Prof. Penny A. Jeggo for critical discussion. The H1299 dA-3 cell line was a generous gift from Dr. Ogiwara. RPE-hTERT cells harboring the OsTIR1 expression cassette (Venegas et al. 2020) was a generous gift from Dr. Kanemaki. We thank Yoshimi Omi, Akiko Shibata, Yoko Hayashi, Yukihiko Yoshimatsu, Itaru Sato, Shiho Nakanishi, Naho Takashima, Hiroko Iino, and Yoshihiko Hagiwara for assisting with the laboratory work and other supports. This work utilized the core research facility of Center for Biology and Integrative Medicine, The University of Tokyo, which was organized by The University of Tokyo Center for NanoBio Integration entrusted by Ministry of Education, Culture, Sports, Science and Technology (MEXT) Japan. This work was carried out under the support of the Isotope Science Center, The University of Tokyo. A part of this study was conducted through the Joint Usage/Research Center Program of the Radiation Biology Center, Kyoto University. The Radiation Biology Center is a joint usage research center certified by MEXT Japan. A.S. is a visiting associate professor of the Radiation Biology Center, Graduate School of Biostudies, Kyoto University. This work was supported by JSPS KAKENHI grant number JP18K18191, JP20KK0339, and JP21H03597 to T.Y. JP15H04902 and JP15K14376 to K.M. JP26701005 and JP17H04713 to A.S. the Takeda Science Foundation to T.Y. and A.S. the Uehara Memorial Foundation to A.S. the Mitsubishi Foundation to A.S. and the Yamada Science Foundation to T.Y. T.Y. R.K. and A.S. conceived and designed the study, discussing it with K.M. The experiments were performed by R.K. M.Y. Y.U. and T.Y. T.Y. R.K. M.Y. T.O. and A.S. analyzed the results. T.Y. L.Z. K.M. and A.S. wrote and edited the manuscript. A.S. supervised the study. The authors declare no competing interests.
Funding Information:
We thank Prof. Penny A. Jeggo for critical discussion. The H1299 dA-3 cell line was a generous gift from Dr. Ogiwara. RPE-hTERT cells harboring the OsTIR1 expression cassette ( Venegas et al., 2020 ) was a generous gift from Dr. Kanemaki. We thank Yoshimi Omi, Akiko Shibata, Yoko Hayashi, Yukihiko Yoshimatsu, Itaru Sato, Shiho Nakanishi, Naho Takashima, Hiroko Iino, and Yoshihiko Hagiwara for assisting with the laboratory work and other supports. This work utilized the core research facility of Center for Biology and Integrative Medicine, The University of Tokyo, which was organized by The University of Tokyo Center for NanoBio Integration entrusted by Ministry of Education, Culture, Sports, Science and Technology (MEXT) Japan. This work was carried out under the support of the Isotope Science Center, The University of Tokyo. A part of this study was conducted through the Joint Usage/Research Center Program of the Radiation Biology Center, Kyoto University. The Radiation Biology Center is a joint usage research center certified by MEXT Japan. A.S. is a visiting associate professor of the Radiation Biology Center, Graduate School of Biostudies, Kyoto University. This work was supported by JSPS KAKENHI grant number JP18K18191 , JP20KK0339 , and JP21H03597 to T.Y., JP15H04902 and JP15K14376 to K.M., JP26701005 and JP17H04713 to A.S., the Takeda Science Foundation to T.Y. and A.S., the Uehara Memorial Foundation to A.S., the Mitsubishi Foundation to A.S., and the Yamada Science Foundation to T.Y.
Publisher Copyright:
© 2022 The Authors
PY - 2022/2/1
Y1 - 2022/2/1
N2 - Single-stranded DNA (ssDNA) arising as an intermediate of cellular processes on DNA is a potential vulnerability of the genome unless it is appropriately protected. Recent evidence suggests that R-loops, consisting of ssDNA and DNA-RNA hybrids, can form in the proximity of DNA double-strand breaks (DSBs) within transcriptionally active regions. However, how the vulnerability of ssDNA in R-loops is overcome during DSB repair remains unclear. Here, we identify RAP80 as a factor suppressing the vulnerability of ssDNA in R-loops, chromosome translocations, and deletions during DSB repair. Mechanistically, RAP80 prevents unscheduled nucleolytic processing of ssDNA in R-loops by CtIP. This mechanism promotes efficient DSB repair via transcription-associated end joining dependent on BRCA1, Polθ, and LIG1/3. Thus, RAP80 suppresses the vulnerability of R-loops during DSB repair, thereby precluding genomic abnormalities in a critical component of the genome caused by deleterious R-loop processing.
AB - Single-stranded DNA (ssDNA) arising as an intermediate of cellular processes on DNA is a potential vulnerability of the genome unless it is appropriately protected. Recent evidence suggests that R-loops, consisting of ssDNA and DNA-RNA hybrids, can form in the proximity of DNA double-strand breaks (DSBs) within transcriptionally active regions. However, how the vulnerability of ssDNA in R-loops is overcome during DSB repair remains unclear. Here, we identify RAP80 as a factor suppressing the vulnerability of ssDNA in R-loops, chromosome translocations, and deletions during DSB repair. Mechanistically, RAP80 prevents unscheduled nucleolytic processing of ssDNA in R-loops by CtIP. This mechanism promotes efficient DSB repair via transcription-associated end joining dependent on BRCA1, Polθ, and LIG1/3. Thus, RAP80 suppresses the vulnerability of R-loops during DSB repair, thereby precluding genomic abnormalities in a critical component of the genome caused by deleterious R-loop processing.
KW - Double-strand break
KW - R-loop
KW - RAP80
KW - Transcription-associated DSB repair
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U2 - 10.1016/j.celrep.2022.110335
DO - 10.1016/j.celrep.2022.110335
M3 - Article
C2 - 35108530
AN - SCOPUS:85123843882
SN - 2211-1247
VL - 38
JO - Cell Reports
JF - Cell Reports
IS - 5
M1 - 110335
ER -