二本鎖ＤＮＡ切断により誘導される複製プログラム変動のメカニズム

• Project/Area Number: 14F03511
• Research Category: Grant-in-Aid for JSPS Fellows
• Allocation Type: Single-year Grants
• Section: 外国
• Research Field: Architectural history/Design
• Research Institution: Tokyo Metropolitan Institute of Medical Science
• Principal Investigator: 正井 久雄 公益財団法人東京都医学総合研究所, ゲノム医科学研究分野, 副所長 (40229349)
• Co-Investigator(Kenkyū-buntansha): LAI MONG SING 公益財団法人東京都医学総合研究所, ゲノム医科学研究分野, 外国人特別研究員 ; LAI Mong Sing 公益財団法人東京都医学総合研究所, ゲノム医科学研究分野, 外国人特別研究員
• Project Period (FY): 2014-04-25 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥2,400,000 (Direct Cost: ¥2,400,000); Fiscal Year 2015: ¥1,200,000 (Direct Cost: ¥1,200,000); Fiscal Year 2014: ¥1,200,000 (Direct Cost: ¥1,200,000)
• Keywords: Rif1 / yKu70 / Double-strand DNA break / DNA replication / Replication origin / Late and dormant origin / Telomere / Budding yeast / Double-strand break / Dormant origins

Outline of Annual Research Achievements

Various types of DNA damage can induce activation of dormant replication origins. Previous studies showed that a single and irrepairable double-strand break (DSB) in the proximity of a dormant replication origin is able to promote origin firing, likely to rescue replication of the broken chromosome fragment (Doksani et al., Cell 2009). In this study, We aim at elucidating the mechanisms leading to dormant origin firing following DSB formation.
Here we showed that DNA damage checkpoints do not counteract DSB-induced dormant origin firing but the dormant origin firing activities are counteracted by histone acetylation state. We also identified telomere-related proteins, Rif1 and Ku complex, as potential regulators of dormant origins even in the absence of DSB. We found that Rif1 and yKu70 bind preferentially at regions between dormant origins and DSB site but not at the other side of the DSB site. The binding profiles of Rif1 and yKu70 at those regions are not affected by DSB formation. Besides, we found that absence of yKu70 prevents Rif1 from binding at regions between dormant origins and DSB site. Our results indicate an important role of Rif1 and yKu70 in controlling dormant origin activity near to DSB site.

Research Progress Status

28年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

28年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] CDC28 phosphorylates Cac1p and regulates the association of Chromatin Assembly Factor I with chromatin. (2015)
  • Author(s): Jeffery, D., Kakusho, N., You, Z., Gharib, M., Wyse, B., Drury, E., Weinreich, M., Thibault, P. Verreault, A., Masai, H. and Yankulov, K.
  • Journal Title: Cell Cycle Volume: 14 Pages: 74-85
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Definition of the transcription factor TdIF1 consensus binding sequence through genome-wide mapping of its binding sites. (2015)
  • Author(s): Kotaro Koiwai, Takashi Kubota, Nobuhisa Watanabe, Katsutoshi Hori, Osamu Koiwai and Hisao Masai
  • Journal Title: Genes to Cells Volume: 20 Issue: 3 Pages: 242-254
  • DOI: 10.1111/gtc.12216
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] NCOA4 Transcriptional Coactivator Inhibits Activation of DNA Replication Origins. (2014)
  • Author(s): Bellelli, R., Castellone, M.D., Guida, T., Limongello, R., Dathan, N.A., Merolla, F., Cirafici, A.M., Affuso, A., Masai, H., Costanzo, V., Grieco, D., Fusco, A., Santoro, M., and Carlomagno, F.
  • Journal Title: Mol. Cell Volume: 55 Issue: 1 Pages: 123-137
  • DOI: 10.1016/j.molcel.2014.04.031
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Regulation and roles of Cdc7 kinase under replication stress. (2014)
  • Author(s): Yamada, M., Masai, H., and Bartek, J.
  • Journal Title: Cell Cycle Volume: 13 Issue: 12 Pages: 1859-1866
  • DOI: 10.4161/cc.29251
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Cell cycle synchronization and flow cytometry analysis of mammalian cell. (2014)
  • Author(s): Yoshizawa-Sugata, N. and Masai, H.
  • Journal Title: Methods in Molecular Biology Volume: 1170 Pages: 279-293
  • DOI: 10.1007/978-1-4939-0888-2_13
  • ISBN: 9781493908875, 9781493908882
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] ATM in prevention of genomic instability. (2014)
  • Author(s): Hisao Masai
  • Journal Title: Cell Cycle Volume: 13 Issue: 6 Pages: 882-883
  • DOI: 10.4161/cc.28216
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Temporal and spatial regulation of eukaryotic DNA replication: from regulated initiation to genome-scale timing program. (2014)
  • Author(s): Renard-Guillet, C., Kanoh, Y., Shirahige, K., and Masai, H.
  • Journal Title: Seminars in Cell & Developmental Biology, Volume: 30 Pages: 110-120
  • DOI: 10.1016/j.semcdb.2014.04.014
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] H2B Mono-ubiquitylation Facilitates Fork Stalling and Recovery during Replication Stress by Coordinating Rad53 Activation and Chromatin Assembly (2014)
  • Author(s): Chia-Yeh Lin, Meng-Ying Wu, Sophie Gay, Lisette Marjavaara, Mong Sing Lai, Wei-Chun Hsiao, Shih-Hsun Hung, Hsin-Yi Tseng, Duncan Edward Wright, Chen-Yi Wang, Guoo-Shyng W. Hsu, Didier Devys, Andrei Chabes, and Cheng-Fu Kao
  • Journal Title: PLoS Genetics Volume: 10 Issue: 10 Pages: 1-17
  • DOI: 10.1371/journal.pgen.1004667
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Replicon dynamics in response to double-strand break formation (2016)
  • Author(s): MongSing Lai
  • Organizer: The 10th 3R Symposium
  • Place of Presentation: ホテル一畑（島根県松江市）
  • Year and Date: 2016-11-13
  • Int'l Joint Research
• [Presentation] 生物種を超えた統一像と多様性 (Conserved mechanisms and diversity of genome DNA replication: from bacteria to human)」イントロダクション (2014)
  • Author(s): 正井 久雄
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川）
  • Year and Date: 2014-11-25 – 2014-11-27
  • Invited
• [Presentation] Mechanisms controlling replicon dynamics following double-strand break formation (2014)
  • Author(s): Lai Mong Sing
  • Organizer: The 9th 3R Symposium
  • Place of Presentation: 御殿場高原リゾート（静岡県）
  • Year and Date: 2014-11-17 – 2014-11-21
  • Invited
• [Presentation] Regulation of mammalian Mcm helicase complexes (2014)
  • Author(s): Hisao Masai, You Zhiying, Koji L. Ode, Naoko Kakusho, Rino Fukatsu, and Haruhiko Takisawa
  • Organizer: 第87回日本生化学会大会シンポジウム「ゲノムを支えるDNAヘリカーゼの動態を探求する新たな研究展開」
  • Place of Presentation: 国立京都国際会館（京都）
  • Year and Date: 2014-10-15 – 2014-10-18
  • Invited
• [Presentation] 染色体DNA複製プログラムの制御機構 (2014)
  • Author(s): 正井 久雄
  • Organizer: 大阪大学蛋白質研究所セミナー「染色体伝承の分子背景：複製から染色体分離まで」
  • Place of Presentation: 大阪大学（大阪）
  • Year and Date: 2014-09-25 – 2014-09-26
  • Invited
• [Presentation] Regulation of DNA replication program in fission yeast and human cells (2014)
  • Author(s): Hisao Masai
  • Organizer: Marco Foiani（Italy, Milan）教授の招聘により IFOM Foundation – F.I.R.C. Institute of Molecular Oncology Foundationで講演
  • Place of Presentation: Italia, Milan
  • Year and Date: 2014-09-16
  • Invited
• [Presentation] Mrc1/Claspinの分子内相互作用による活性制御および新規機能 (2014)
  • Author(s): 正井 久雄
  • Organizer: 九州大学 大学院薬学研究院 招聘研究セミナー
  • Place of Presentation: 九州大学（福岡）
  • Year and Date: 2014-08-29
  • Invited
• [Presentation] Regulation of replication program in fission yeast and human cells. (2014)
  • Author(s): Hisao Masai
  • Organizer: Peking University（特別招聘講演）
  • Place of Presentation: Peking University（中国）
  • Year and Date: 2014-05-16
  • Invited
• [Book] “DNA replication: From Old Principles to New Discovery” in “Advances in Experimental Medicine and Biology” (2017)
  • Author(s): Kenji Moriyama, Mong Sing Lai, and Hisao Masai
  • Publisher: Springer
• [Book] DNA Replication, Recombination and Repair: Molecular Mechanisms and Pathology (2015)
  • Author(s): Motoshi Hayano, Seiji Matsumoto and Hisao Masai
  • Publisher: Springer
• [Remarks] 公益財団法人東京都医学総合研究所ゲノム動態プロジェクト
  • URL: http://www.igakuken.or.jp/genome/