Genomic instability by Topoisomerase 1 induced by AID-dependent RNA editing

• Project/Area Number: 25440007
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Molecular biology
• Research Institution: Kyoto University
• Principal Investigator: Kobayashi Maki 京都大学, 医学(系)研究科(研究院), 准教授 (20400690)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2015: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2013: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
• Keywords: トポイソメラーゼ１ / AID / ゲノム不安定性 / 免疫グロブリン遺伝子 / 遺伝子不安定性 / Topoisomerase1 / miRNA / Ago2

Outline of Final Research Achievements

In order to identify the responsive site in Topoisomerase 1 (Top1) mRNA to AID, Top1 3'UTR knockout cells were generated from CH12 B lymphoma cells. Unexpectedly class switch recombination in these knockout cells was not abolished supposing that coding region of Top1 mRNA is the target of AID's regulation.
On the other hand, Top1's involvement in DNA cleavages in immunoglobulin (Ig) gene was further clarified by the proteasome inhibitor and the novel Top1 inhibitors. Additionally by Top1-GFP fusion protein and trapping by GFP, it is elucidated that FACT enrolls as an adopter molecule between Top1 and H3K4me3, modified histone with help of BRG1.

Research Products

• [Journal Article] Chromatin remodeler SMARCA4 recruits topoisomerase 1 and suppresses transcription associated genomic instability (2016)
  • Author(s): Afzal Husain, Nasim A. Begum, Takako Taniguchi, Hisaaki Taniguchi, Maki Kobayashi, and Tasuku Honjo.
  • Journal Title: Nat. Commu. Volume: 7 Issue: 1 Pages: 00-00
  • DOI: 10.1038/ncomms10549
  • NAID: 120006535056
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] LSD1/KDM1A promotes hematopoietic commitment of hemangioblasts through downregulation of Etv2. (2015)
  • Author(s): Takeuchi, M., Fuse, Y., Watanabe, M., Andrea, C.S., Takeuchi, M., Nakajima, H., Ohashi, K., Kaneko, H., Kobayashi-Osaki, M., Yamamoto, M. and Kobayashi, M.
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 112 Issue: 45 Pages: 13922-13927
  • DOI: 10.1073/pnas.1517326112
  • NAID: 120007135765
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] APE1 is dispensable for S region cleavage but required for its repair in class switch recombination (2014)
  • Author(s): Jianliang Xu, Afzal Husain, Wenjun Hu, Tasuku Honjo and Maki Kobayashi.
  • Journal Title: PNAS USA Volume: 111 Issue: 48 Pages: 17242-17247
  • DOI: 10.1073/pnas.1420221111
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] C-terminal region of activation-induced cytidine deaminase (AID) is required for efficient class switch recombination and gene conversion. (2014)
  • Author(s): Sabouri S, Kobayashi M, Begum NA, Xu J, Hirota K, Honjo T
  • Journal Title: Proc Natl Acad Sci U S A. Volume: 111 Issue: 6 Pages: 2253-2258
  • DOI: 10.1073/pnas.1324057111
  • NAID: 120005395696
  • Peer Reviewed
• [Journal Article] Activation-induced cytidine deaminase is dispensable for virus-mediated liver and skin tumor development in mouse models. (2014)
  • Author(s): Nguyen T, Xu J, Chikuma S, Hiai H, Kinoshita K, Moriya K, Koike K, Marcuzzi GP, Pfister H, Honjo T, Kobayashi M.
  • Journal Title: Int Immunol Volume: 26 Issue: 7 Pages: 397-406
  • DOI: 10.1093/intimm/dxu040
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Activation-induced cytidine deaminase is dispensable for virus-mediated liver and skin tumor development in mouse models in press (2014)
  • Author(s): Tung Nguyen,Jianliang Xu,Shunsuke Chikuma,Hiroshi Hiai,Kazuo Kinoshita,Kyoji Moriya,Kazuhiko Koike,Gian Paolo Marcuzzi, Herbert Pfister, Tasuku Honjo, and Maki Kobayashi
  • Journal Title: Int Immunol Volume: 未定
  • Peer Reviewed
• [Journal Article] GATA factor switching from GATA2 to GATA1 contributes to erythroid differentiation (2013)
  • Author(s): Suzuki M, Kobayashi-Osaki M, Tsutsumi S, Pan X, Ohmori S, Takai J, Moriguchi T, Ohneda O, Ohneda K, Shimizu R, Kanki Y, Kodama T, Aburatani H, Yamamoto M
  • Journal Title: Genes Cells Volume: 18 Issue: 11 Pages: 921-933
  • DOI: 10.1111/gtc.12086
  • Peer Reviewed
• [Journal Article] In vivo analysis of Aicda gene regulation: a critical balance between upstream enhancers and intronic silencers governs appropriate expression (2013)
  • Author(s): Le Thi Huong, Maki Kobayashi,et al
  • Journal Title: PLoSONE Volume: vol.8 Issue: 4 Pages: e61433-e61433
  • DOI: 10.1371/journal.pone.0061433
  • Peer Reviewed
• [Presentation] Immunoglobulin gene diversification by activation-induced cytidine deaminase (AID) through Topoisomerase1 (Top1). (2015)
  • Author(s): Maki Kobayashi, Hiroyuki Wakaguri, Masakazu Shimizu, Koh-ichiro Higasa, Fumihiko Matsuda and Tasuku Honjo
  • Organizer: The 2nd IFOM-Kyoto University Joint Symposium
  • Place of Presentation: 京都大学芝蘭会館
  • Year and Date: 2015-10-06
  • Int'l Joint Research
• [Presentation] トポイソメラーゼ１を介したAIDによる免疫グロブリン遺伝子の多様化 (2015)
  • Author(s): 小林 牧、本庶 佑
  • Organizer: 第17回RNA meeting
  • Place of Presentation: 札幌パークホテル
  • Year and Date: 2015-07-15
• [Presentation] Immunoglobulin gene diversification by activation-induced cytidine deaminase (AID) through Topoisomerase1 (Top1) . (2015)
  • Author(s): Maki Kobayashi, Jianliang Xu, Tasuku Honjo
  • Organizer: The Golden Anniversary of B Cells Discovery, Keystone Symposia
  • Place of Presentation: Fairmont Banff Springs, Banff, Canada
  • Year and Date: 2015-03-22 – 2015-03-26
• [Presentation] AID-regulated Topoisomerase1 is the DNA cleaving enzyme during immunoglobulin diversification (2014)
  • Author(s): Maki Kobayashi
  • Organizer: Kyoto University & National Taiwan University Symposium
  • Place of Presentation: 京都大学
  • Year and Date: 2014-09-01
  • Invited
• [Presentation] AID regulates Top1 translation by changing miRNA loading to Top1 mRNA (2014)
  • Author(s): Maki Kobayashi, Masayuki Shimizu, Koichiro Higasa, Fumihiko Matsuda and Tasuku Honjo
  • Organizer: Frontiers in Immunology 2014
  • Place of Presentation: Novel Forum,Karolinska Institutet, Stockholm, Sweden
  • Year and Date: 2014-06-18 – 2014-06-21
• [Remarks] 「AID-C末端によるDNAシナプス形成」について
  • URL: http://www.kyoto-u.ac.jp/ja/news_data/h/h1/news6/2013_1/140128_2.htm