TY - JOUR
T1 - Replication stress induces accumulation of FANCD2 at central region of large fragile genes
AU - Okamoto, Yusuke
AU - Iwasaki, Watal M.
AU - Kugou, Kazuto
AU - Takahashi, Kazuki K.
AU - Oda, Arisa
AU - Sato, Koichi
AU - Kobayashi, Wataru
AU - Kawai, Hidehiko
AU - Sakasai, Ryo
AU - Takaori-Kondo, Akifumi
AU - Yamamoto, Takashi
AU - Kanemaki, Masato T.
AU - Taoka, Masato
AU - Isobe, Toshiaki
AU - Kurumizaka, Hitoshi
AU - Innan, Hideki
AU - Ohta, Kunihiro
AU - Ishiai, Masamichi
AU - Takata, Minoru
N1 - Funding Information:
JSPS KAKENHI [JP23114010 to M.T., JP26550026 to M.T., JP15H01738 to M.T., JP26281021 to M.I., JP25116002 to H.K., JP17H01408 to H.K.]; Ministry of Health, Labour and Welfare (to M.T.); Uehara Memorial Foundation (to M.T.); Waseda University (to H.K.). Funding for open access charge: Uehara Memorial Foundation (to M.T.). Conflict of interest statement. None declared.
Funding Information:
The authors would like to thank Dr James Hejna (Kyoto University) for critical reading of the manuscript; Drs Patrick Calsou, Daniel Voytas, Rudolf Jaenisch, Hiroyuki Miyoshi, Makoto Nakanishi and Keith Joung for reagents; Drs Yasutoshi Agata (Shiga University of Medical Science), Naoyuki Kataoka (The University of Tokyo), Daisuke Kaida (University of Toyama) for advice in ChIP procedure; Drs Mari Miyaji, Kimiko Tsutsui and Ken Tsutsui (Okayama University Medical School), or Nozomi Sugimoto, and Masatoshi Fujita (Kyushu University) for discussions; technical and secretarial help from Chinatsu Ohki, Akiko Watanabe, Seiko Arai, Ayumi Katayama, and Fan Peng. The Radiation Biology Center, Kyoto University, is a Joint Usage Research Center supported by the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan. A part of the authors’ work has been performed in collaborative research projects in the Joint Usage Research Center. The authors declare no competing financial interests. JSPS KAKENHI [JP23114010 to M.T., JP26550026 to M.T., JP15H01738 to M.T., JP26281021 to M.I., JP25116002 to H.K., JP17H01408 to H.K.]; Ministry of Health, Labour and Welfare (to M.T.); Uehara Memorial Foundation (to M.T.); Waseda University (to H.K.). Funding for open access charge: Uehara Memorial Foundation (to M.T.).
Funding Information:
The authors would like to thank Dr James Hejna (Kyoto University) for critical reading of the manuscript; Drs Patrick Calsou, Daniel Voytas, Rudolf Jaenisch, Hi-royuki Miyoshi, Makoto Nakanishi and Keith Joung for reagents; Drs Yasutoshi Agata (Shiga University of Medical Science), Naoyuki Kataoka (The University of Tokyo), Daisuke Kaida (University of Toyama) for advice in ChIP procedure; Drs Mari Miyaji, Kimiko Tsutsui and Ken Tsutsui (Okayama University Medical School), or Nozomi Sugimoto, and Masatoshi Fujita (Kyushu University) for discussions; technical and secretarial help from Chinatsu Ohki, Akiko Watanabe, Seiko Arai, Ayumi Katayama, and Fan Peng. The Radiation Biology Center, Kyoto University, is a Joint Usage Research Center supported by the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan. A part of the authors’ work has been performed in collaborative research projects in the Joint Usage Research Center. The authors declare no competing financial interests.
Publisher Copyright:
© The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research.
PY - 2018/4/6
Y1 - 2018/4/6
N2 - During mild replication stress provoked by low dose aphidicolin (APH) treatment, the key Fanconi anemia protein FANCD2 accumulates on common fragile sites, observed as sister foci, and protects genome stability. To gain further insights into FANCD2 function and its regulatory mechanisms, we examined the genome-wide chromatin localization of FANCD2 in this setting by ChIP-seq analysis. We found that FANCD2 mostly accumulates in the central regions of a set of large transcribed genes that were extensively overlapped with known CFS. Consistent with previous studies, we found that this FANCD2 retention is R-loop-dependent. However, FANCD2 monoubiquitination and RPA foci formation were still induced in cells depleted of R-loops. Interestingly, we detected increased Proximal Ligation Assay dots between FANCD2 and R-loops following APH treatment, which was suppressed by transcriptional inhibition. Collectively, our data suggested that R-loops are required to retain FANCD2 in chromatin at the middle intronic region of large genes, while the replication stress-induced upstream events leading to the FA pathway activation are not triggered by R-loops.
AB - During mild replication stress provoked by low dose aphidicolin (APH) treatment, the key Fanconi anemia protein FANCD2 accumulates on common fragile sites, observed as sister foci, and protects genome stability. To gain further insights into FANCD2 function and its regulatory mechanisms, we examined the genome-wide chromatin localization of FANCD2 in this setting by ChIP-seq analysis. We found that FANCD2 mostly accumulates in the central regions of a set of large transcribed genes that were extensively overlapped with known CFS. Consistent with previous studies, we found that this FANCD2 retention is R-loop-dependent. However, FANCD2 monoubiquitination and RPA foci formation were still induced in cells depleted of R-loops. Interestingly, we detected increased Proximal Ligation Assay dots between FANCD2 and R-loops following APH treatment, which was suppressed by transcriptional inhibition. Collectively, our data suggested that R-loops are required to retain FANCD2 in chromatin at the middle intronic region of large genes, while the replication stress-induced upstream events leading to the FA pathway activation are not triggered by R-loops.
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U2 - 10.1093/nar/gky058
DO - 10.1093/nar/gky058
M3 - Article
C2 - 29394375
AN - SCOPUS:85052072855
VL - 46
SP - 2932
EP - 2944
JO - Nucleic Acids Research
JF - Nucleic Acids Research
SN - 0305-1048
IS - 6
ER -