Identification and functional analysis of novel vertebrate chromosomal proteins with using Multi-Classifier Combinational Proteomics

• Project/Area Number: 25870487
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Cell biology; Molecular biology
• Research Institution: Kochi University
• Principal Investigator: Ohta Shinya 高知大学, 教育研究部医療学系基礎医学部門, 特任助教 (00631194)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 染色体 / 分裂期 / プロテオミクス

Outline of Final Research Achievements

In this study we identified CENP-32. CENP-32 appears to be required for centrosomes to integrate into a fully functional spindle that notonly nucleates astral microtubules, but also is able to nucleate and bind to kinetochore andcentral spindle microtubules. Additional data suggest that NuMA tethers microtubules at the anastral spindle poles and that augmin is required for centrosome detachment after CENP-32 depletion, possibly due to an imbalance of forces within the spindle.
Next we identified CSAP and proposed that CSAP associates with MTs around centrosomes to stabilize MTs during mitosis, ensuring proper bipolar spindle formation and maintenance.

Research Products

• [Int'l Joint Research] エジンバラ大学(英国)
• [Journal Article] Auxin/AID versus conventional knockouts: distinguishing the roles of CENP-T/W in mitotic kinetochore assembly and stability. (2016)
  • Author(s): Laura Wood, Daniel G. Booth, Giulia Vargiu, Shinya Ohta, Flavia deLima Alves, Kumiko Samejima, Tatsuo Fukagawa, Juri Rappsilber, and William C Earnshaw
  • Journal Title: Open Biol. Volume: 6(1) Pages: 150230-150230
  • DOI: 10.1098/rsob.150230
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Polyglutamylated tubulin binding protein C1orf96/CSAP is involved in microtubule stabilization in mitotic spindles (2015)
  • Author(s): Shinya Ohta, Mayako Hamada, Nobuko Sato, Iyo Toramoto
  • Journal Title: PLOS ONE Volume: 10(11)
  • DOI: 10.1371/journal.pone.0142798
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [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: Mol Biol Cell Volume: 26(7) Pages: 1225-1237
  • DOI: 10.1091/mbc.e14-09-1366
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] 曖昧なプロテオミクス(Fuzzy Proteomics)による分裂期染色体構造の決定 (2013)
  • Author(s): 太田 信哉
  • Journal Title: The lung Volume: 21(3) Pages: 294-299
• [Presentation] プロテオミクスによる分裂期クロマチン制御構造の解析 (2014)
  • Author(s): 太田 信哉
  • Organizer: 第32回染色体ワークショップ、第13回核ダイナミクス研究会
  • Place of Presentation: 広島 (安芸グランドホテル)
  • Year and Date: 2014-12-12 – 2014-12-14
• [Presentation] 分裂期染色体上におけるタンパク質リン酸化の網羅的解析 (2014)
  • Author(s): 太田 信哉, 高木 俊輔, 木村 迪子, 石濱 泰
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: 横浜 (パシフィコ横浜)
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] Defining the structure of mitotic chromosomes using multi-classifier combinatorial proteomics together with DT40 genetics (2013)
  • Author(s): Shinya Ohta, Laura Wood, Jimi-Carlo Bukowski-Wills, Itaru Samejima Luis Fernando Montano, Juri Rappsilber and William C. Earnshaw
  • Organizer: 12th Human Proteome Organisation World Congress
  • Place of Presentation: パシフィコ横浜（横浜）
• [Presentation] Proteomics study of vertebrate mitotic chromosomes. (2013)
  • Author(s): Itaru Samejima, Flavia de Lima Alves, Shinya Ohta, Juri Rappsilber, Tatsuo Fukagawa and William C Earnshaw
  • Organizer: Gordon Research Conference – chromosome dynamics
  • Place of Presentation: ルネッサンストスカーニャホテル（ルッカ イタリア）
• [Presentation] 多次元プロテオミクスによる分裂期染色体の制御構造解析 (2013)
  • Author(s): 太田 信哉, Luis Fernando Montaño, Laura Wood, Itaru, Samejima, Flavia de Lima Alves, Juri Rapsilber, William C Earnshaw
  • Organizer: 第64回日本染色体学会年会
  • Place of Presentation: 富山大学（富山）
• [Presentation] プロテオミクスと遺伝学の融合による分裂期染色体の制御構造解析 (2013)
  • Author(s): 太田 信哉, Laura Wood, Juri Rappsilber, William C. Earnshaw
  • Organizer: 第85回日本遺伝学会年会
  • Place of Presentation: 慶応大学（横浜）
• [Presentation] 多次元プロテオミクスを用いた分裂期染色体構造の解析 (2013)
  • Author(s): 太田 信哉
  • Organizer: 第2回JHUPOサテライトシンポジウム
  • Place of Presentation: 愛媛大学（松山）