セントロメアサイズを規定する分子機構の解析

• Project Area: Functions of non-coding DNA region for genome integrity
• Project/Area Number: 26114715
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Osaka University (2015); National Institute of Genetics (2014)
• Principal Investigator: 深川 竜郎 大阪大学, 生命機能研究科, 教授 (60321600)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Discontinued (Fiscal Year 2015)
• Budget Amount: ¥10,400,000 (Direct Cost: ¥8,000,000、Indirect Cost: ¥2,400,000); Fiscal Year 2015: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2014: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
• Keywords: セントロメア / クロマチン / CENP-A / エピジェネティックス

Outline of Annual Research Achievements

染色体分配に必須なセントロメアは、動原体が形成されるゲノム領域であり、典型的な機能を持った非コードDNA領域である。したがってセントロメアのゲノム領域を研究することで、非コードDNA領域の理解に貢献できる。我々は、これまでに、LacO-LacIのシステムと染色体工学を併用して人工動原体を作成する過程で、予想外にもセントロメアのサイズが通常の3~5倍程度に巨大化している細胞を見いだした。本研究では、このセントロメアのサイズが大きくなった細胞を利用して、「セントロメア領域のサイズを規定する分子機構」の解明を目的とする。この目的の達成のために、26年度は、セントロメアを巨大化させる因子の同定を試み、HJURPタンパク質の255-500アミノ酸の領域にセントロメアを巨大化させる機能があることを見出した。また、巨大化したセントロメアをもつ染色体の動態を解析した結果、通常の染色体とその挙動が変わらないことを明らかにできた。27年度において、HJURPの255-500アミノ酸の領域と結合するタンパク質としてKNL2を同定し、KNL2とHJURPの結合がセントロメア巨大化に重要であることを明らかにした。さらに、通常の細胞においても、HJURPを過剰発現することで、セントロメアが巨大することも示すことができた。

Research Progress Status

27年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

27年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] CENP-32 is required to maintain centrosomal dominance in bipolar spindle assembly (2015)
  • Author(s): Shinya Ohta, Laura Wood, Iyo Toramoto, Ken-Ichi Yagyu, Tatsuo Fukagawa, and William C. Earnshaw
  • Journal Title: Molecular Biology of the Cell. Volume: 26(7) Issue: 7 Pages: 1225-1237
  • DOI: 10.1091/mbc.e14-09-1366
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] HJURP is involved in the expansion of centromeric chromatin. (2015)
  • Author(s): Perpelescu M, Hori T, Toyoda A, Misu S, Monma N, Ikeo K, Obuse C, Fujiyama A, Fukagawa T.
  • Journal Title: Mol Biol Cell Volume: 26 Pages: 2742-2754
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Cell Division: A New Role for the Kinetochore in Central Spindle Assembly. (2015)
  • Author(s): Fukagawa T.
  • Journal Title: Curr Biol. Volume: 25 Issue: 13 Pages: 554-557
  • DOI: 10.1016/j.cub.2015.05.016
  • Peer Reviewed / Open Access
• [Journal Article] Dynamic changes in the CCAN organization through CENP-C during cell-cycle progression. (2015)
  • Author(s): Nagpal H, Hori T, Furukawa A, Sugase K, Osakabe A, Kurumizaka H, Fukagawa T.
  • Journal Title: Mol Biol Cell Volume: 26
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Crystal structure and stable property of the cancer-associated heterotypic nucleosome containing CENP-A and H3.3. (2014)
  • Author(s): Arimura Y, Shirayama K, Horikoshi N, Fujita R, Taguchi H, Kagawa W, Fukagawa T, Almouzni G, Kurumizaka H.
  • Journal Title: Sci Rep. Volume: 4 Issue: 1 Pages: 7115-7115
  • DOI: 10.1038/srep07115
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Neocentromeres. (2014)
  • Author(s): Fukagawa T, Earnshaw WC.
  • Journal Title: Curr Biol Volume: 24 Issue: 19 Pages: R946-R947
  • DOI: 10.1016/j.cub.2014.08.032
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] The centromere: chromatin foundation for the kinetochore machinery. (2014)
  • Author(s): Fukagawa T, Earnshaw WC.
  • Journal Title: Dev Cell Volume: 30 Issue: 5 Pages: 496-508
  • DOI: 10.1016/j.devcel.2014.08.016
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Reorganization of Damaged Chromatin by the Exchange of Histone Variant H2A.Z-2. (2014)
  • Author(s): Nishibuchi I
  • Journal Title: Int J Radiat Oncol Biol Phys. Volume: 89 Issue: 4 Pages: 736-744
  • DOI: 10.1016/j.ijrobp.2014.03.031
  • Peer Reviewed / Open Access
• [Journal Article] Histone H4 Lys 20 monomethylation of the CENP-A nucleosome is essential for kinetochore assembly. (2014)
  • Author(s): Hori, T., Shang, W.H., Toyoda, A., Misu, S., Monma, N., Ikeo, K., Molina, O., Vargiu, G., Fujiyama, A., Kimura, H., Earnshaw, W.C., Fukagawa, T.
  • Journal Title: Dev Cell Volume: 29 Issue: 6 Pages: 740-749
  • DOI: 10.1016/j.devcel.2014.05.001
  • Peer Reviewed / Open Access
• [Journal Article] The histone-fold complex MHF is remodeled by FANCM to recognize branched DNA and protect genome stability. (2014)
  • Author(s): Fox D 3rd, Yan Z, Ling C, Zhao Y, Lee DY, Fukagawa T, Yang W, Wang W.
  • Journal Title: Cell Res Volume: 24 Issue: 5 Pages: 560-575
  • DOI: 10.1038/cr.2014.42
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Meiosis-specific cohesin mediates homolog recognition in mouse spermatocytes (2014)
  • Author(s): Ishiguro KI., Kim J., Shibuya H., Hernandez-Hernandez A., Suzuki A., Fukagawa T., Shioi G., Kiyonari H., Li XC., Schimenti J., Hoog C., Watanabe Y.
  • Journal Title: Genes Dev. Volume: 28 Issue: 6 Pages: 594-607
  • DOI: 10.1101/gad.237313.113
  • Peer Reviewed
• [Journal Article] The CENP-O complex requirement varies among different cell types. (2014)
  • Author(s): Kagawa, N., Hori, T., Hoki, Y., Hosoya, O., Tsutsui, K., Saga, Y., Sado, T., and *Fukagawa, T.
  • Journal Title: Chrom. Research Volume: in press Issue: 3 Pages: 293-303
  • DOI: 10.1007/s10577-014-9404-1
  • Peer Reviewed / Open Access
• [Presentation] Specific features of centromeric chromatin for kinetochore assembly (2015)
  • Author(s): Tatsuo Fukagawa
  • Organizer: International Conference on Chromatin Regulation in Proliferation and Differentiation
  • Place of Presentation: Essen, Germany
  • Year and Date: 2015-09-02
  • Invited
• [Presentation] Specific features of centromeric chromatin (2015)
  • Author(s): Tatsuo Fukagawa
  • Organizer: International Symposium on Non-coding DNA and Chromosome Integrity
  • Place of Presentation: Awaji, Japan
  • Year and Date: 2015-08-07
  • Invited
• [Presentation] Specific features of centromeric chromatin (2015)
  • Author(s): Tatsuo Fukagawa
  • Organizer: FASEB meeting on Mitosis: Spindle assembly and function
  • Place of Presentation: Big Sky, Montana, USA
  • Year and Date: 2015-06-21
  • Invited
• [Presentation] Centromere specification and kinetochore assembly (2014)
  • Author(s): Tatsuo Fukagawa
  • Organizer: 3R symposium
  • Place of Presentation: Gotenba (時のすみか)
  • Year and Date: 2014-11-17 – 2014-11-21
  • Invited
• [Presentation] Structural basis of centromeres (2014)
  • Author(s): Tatsuo Fukagawa
  • Organizer: 日本癌学会
  • Place of Presentation: 横浜 (パシフィコ横浜)
  • Year and Date: 2014-09-25 – 2014-09-27
  • Invited
• [Presentation] Centromere Specification and Assembly in Vertebrates (2014)
  • Author(s): Tatsuo Fukagawa
  • Organizer: Gordon Reseach Conference: Centromere Biology
  • Place of Presentation: Boston, USA
  • Year and Date: 2014-07-27 – 2014-08-01
  • Invited
• [Presentation] Phosphorylation of CENP-T controls kinetochore assembly (2014)
  • Author(s): Tatsuo Fukagawa
  • Organizer: タンパク質科学会年会
  • Place of Presentation: 横浜 (パシフィコ横浜)
  • Year and Date: 2014-06-25 – 2014-06-27
  • Invited
• [Presentation] Centromere specific chromatin structure (2014)
  • Author(s): Tatsuo Fukagawa
  • Organizer: IIAS Reseach Conference 2014 Chromatin Decoding
  • Place of Presentation: Kyoto (国際高等研究所)
  • Year and Date: 2014-05-12 – 2014-05-15
  • Invited
• [Remarks] 染色体のセントロメア形成に関わる分子スイッチを発見
  • URL: http://www.nig.ac.jp/Research-Highlights/1468/1527.html
• [Remarks] セントロメアを構成するクロマチン構造
  • URL: http://www.nig.ac.jp/Research-Highlights/1468/1559.html