Mechanism of the replication stress response of variable replication fork complexes

• Project/Area Number: 23570243
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Cell biology
• Research Institution: Tokyo Metropolitan Institute of Medical Science
• Principal Investigator: YOSHIZAWA=SUGATA Naoko 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, 主任研究員 (30344071)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2013: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2012: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2011: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: ゲノム不安定性 / 多能性幹細胞 / 初期胚 / 複製フォーク / 複製ストレス / ES細胞 / AND-1 / Tipin / Rif1 / クロマチン免疫沈降 / 胎生幹細胞 / Timeless / DNA修復 / 未分化

Research Abstract

In attempts to identify fork complex components and its binding proteins, we purified Timeless complexes and found that it specifically contains phosphorylated Mcm4 proteins in the chromatin fraction. We also found that the replication fork proteins AND-1 and Tipin, and a replication timing regulator Rif1, are highly expressed in the undifferentiated mouse embryonic stem (ES) cells. Among them, Rif1 is highly correlated with the undifferentiated states of ES cells. Interestingly, Rif1 may play dual roles in ES cells; In addition to its counteraction with spontaneous differentiation, Rif1 also inhibits the expression of 2 cell-specific genes, which are involved in promoting reprogamming for pluripotency. We found that Rif1 binds to the promoter of differentiation-regulating genes.

Research Products

• [Journal Article] Cell cycle synchronization and flow cytometry analysis of mammalian cells (2014)
  • Author(s): Yoshizawa-Sugata, N. and Masai, H.
  • Journal Title: Methods in Molecular Biology Volume: in press
  • Peer Reviewed
• [Journal Article] Role of multifunctional transcription factor TFII-I and putative tumour suppressor DBC1 in cell cycle and DNA double strand damage repair (2013)
  • Author(s): Tanikawa M, Wada Hiraike O, Yoshizawa Sugata N , Shirane A, Hirano M, Hiraike H, Miyamoto Y, Sone K, Ikeda Y, Kashiyama T, Oda K, Kawana K, Katakura Y, Yano T, Masai H, Roy AL, Osuga Y, Fujii T
  • Journal Title: Br. J. Cancer Volume: 109 Pages: 3042-8
  • Peer Reviewed
• [Journal Article] Role of multifunctional transcription factor TFII-I and putative tumour suppressor DBC1 in cell cycle and DNA double strand damage repair. (2013)
  • Author(s): Tanikawa, M., Wada-Hiraike, O., Yoshizawa-Sugata, N., Shirane A, Hirano M, Hiraike H, Miyamoto, Y., Sone, K., Ikeda, Y., Kashiyama, T., Oda K, Kawana K, Katakura Y, Yano T, Masai, H., Roy AL, Osuga, Y. and Fujii, T.
  • Journal Title: Br. J. Cancer. Volume: 109 Issue: 12 Pages: 3042-3048
  • DOI: 10.1038/bjc.2013.532
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] EBNA1-dependent recruitment of Orc on OriP of Epstein-Barr virus with purified proteins : Stimulation by Cdc6 through Its direct interaction with EBNA1 (2012)
  • Author(s): Moriyama K, Yoshizawa Sugata N, Obuse C, Tsurimoto T, and Masai H
  • Journal Title: Biological Chemistry Volume: 287 Pages: 23977-23994
  • Peer Reviewed
• [Journal Article] Epstein-Barr nuclear antigen 1 (EBNA1)-dependent recruitment of origin recognition complex(Orc) on oriP of Epstein-Barr virus with purified proteins:stimulation by Cdc6 through its direct interaction with EBNA1. (2012)
  • Author(s): Moriyama K
  • Journal Title: The Journal of biological chemistry Volume: 287 Issue: 28 Pages: 23977-23994
  • DOI: 10.1074/jbc.m112.368456
  • Peer Reviewed
• [Presentation] Rif1のES細胞での未分化能維持、および体細胞のリプログラミングにおける機能解析、クロマチン構造制御因子としての可能性 (2013)
  • Author(s): 吉沢直子, 正井久雄
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸(ポスター発表)
• [Presentation] Rif1のES 細胞での未分化能維持、および体細胞のリプログラミングにおける機能解析:クロマチン構造制御因子としての可能性 (2013)
  • Author(s): 吉沢 直子,正井 久雄
  • Organizer: 第36回日本分子生物学会
  • Place of Presentation: 神戸
• [Presentation] Rif1はクロマチン高次構築を制御してES細胞特異的な核内構造を形成する (2012)
  • Author(s): 吉沢直子, 覚正直子, 山崎聡, 深津理乃, 正井久雄
  • Organizer: 第35回日本分子生物学会年会ワークショップ
  • Place of Presentation: 福岡(口頭発表およびポスター発表)
• [Presentation] The maintenance of undifferentiated state in mouse embryonic stem cells by Rap1 interacting factor-1 (Rif1), a potential regulator of chromatin architecture in nuclei (2012)
  • Author(s): Yoshizawa Sugata N、Kakusho N, Yamazaki S, Fukatsu R, and Masai H
  • Organizer: 分子生物学会年会
  • Place of Presentation: 横浜(口頭発表およびポスター)
• [Presentation] The maintenance of undifferentiated state in mouse embryonic stem cells by Rap1 interacting factor-1 (Rif1), a potential regulator of chromatin architecture in nuclei (2012)
  • Author(s): Yoshizawa, N., Kakusho, N., Yamazaki, S., Fukatsu, R. and Masai, H.
  • Organizer: 日本分子生物学会
  • Place of Presentation: 神奈川県横浜市
• [Presentation] Regulation of replication program in fission yeast and human cells (2012)
  • Author(s): Hisao Masai, Satoshi Yamazaki, Yutaka Kanoh, Motoshi Hayano, Seiji Matsumoto, Naoko Yoshizawa, Rino Fukatsu, Naoko Kakusho, and Michie Shimmoto
  • Organizer: 8th 3R meeting
  • Place of Presentation: 兵庫県淡路島
• [Presentation] Cell cycle regulation of mouse embryonic stem cells: E2F- and Emi1-mediated positive feedback regulation (2012)
  • Author(s): Hisao Masai, Hiroko Fujii, Naoko Kakusho, Sayuri Ito, Satoshi Yamazaki, and Naoko Yoshizawa
  • Organizer: 4th Southeast Stem Cell Consortium Workshop at Florida State University
  • Place of Presentation: アメリが合衆国、フロリダ州
• [Presentation] The maintenance of pluripotency in mouse embryonic stem cells by Rif1, a potential regulator of replication domains (2011)
  • Author(s): Yoshizawa Sugata N、Kakusho N, Yamazaki S, Fukatsu R, and Masai H
  • Organizer: Joint CSH-Asia / ISSCR Conference on Cellular Programs & Reprogramming
  • Place of Presentation: 中国,蘇州(ポスター発表)
• [Presentation] The maintenance of pluripotency in mouse embryonic stem cells by Rif1, a potential regulator of replication domains (2011)
  • Author(s): Yoshizawa,N., Kakusho, N., Yamazaki, S., Fukatsu, R. and Masai, H.
  • Organizer: Joint CSH-Asia / ISSCR Conference on Cellular Programs & Reprogramming
  • Place of Presentation: 中国・蘇州
• [Presentation] The maintenance of undifferentiated state in mouse embryonic stem cells by Rap1 interacting factor-1 (Rif1), a potential regulator of chromatin architecture in nuclei (2011)
  • Author(s): Yoshizawa,N., Kakusho, N., Yamazaki, S., Fukatsu, R. and Masai, H.
  • Organizer: 日本分子生物学会
  • Place of Presentation: 横浜
• [Book] Methods in Molecular Biology (2014)
  • Author(s): Yoshizawa-Sugata, N, and Masai.
  • Publisher: Cell cycle synchronization and flow cytometry analysis of mammalian cells
• [Remarks] ホームページ等
  • URL: http://www.igakuken.or.jp/genome/
• [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/