Analysis of novel centrosome regulation in human NSC

• Project/Area Number: 25460376
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: Kanazawa Medical University
• Principal Investigator: ISHIGAKI Yasuhito 金沢医科大学, 総合医学研究所, 教授 (20232275)
• Co-Investigator(Renkei-kenkyūsha): NAKAMURA Yuka 金沢医科大学, 総合医学研究所, 助手 (00565632) ; TOMOSUGI Naohisa 金沢医科大学, 総合医学研究所, 教授 (80155580)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: Y14 / 中心体 / リン酸化 / PhosTagゲル / RBM8A(Y14) / M期 / アポトーシス / RBM8A（Y14)

Outline of Final Research Achievements

Y14-Magohcomplex is a component of the exon junction complex (EJC) required for mRNA metabolism. However, the detailed status and mechanism of the phosphorylation of Y14 is poorly understood. We analyzed in detail Y14 phosphorylation in human cells. Phos-tag gels revealed that the majority of endogenous Y14 was phosphorylated throughout the cell-cycle progression. Nuclear and cytoplasmic Y14 and Y14 in the EJC was also found to be mostly phosphorylated. We also screened the phosphorylated serine by mutational analysis using Phos-tag gels to reveal modifications of serine residues 166 and 168. A single substitution at position 168 that concomitantly abolished the phosphorylation of serine 166 suggested the priority of kinase reaction between these sites. Furthermore, analysis of the role of the binding protein Magoh in Y14 phosphorylation revealed its inhibitory effect in vitro and in vivo.

Research Products

• [Journal Article] Nonsense-mediated mRNA decay factor Upf2 exists in both the nucleoplasm and the cytoplasm. (2016)
  • Author(s): Tatsuno T, Nakamura Y, Ma S, Tomosugi N, Ishigaki Y
  • Journal Title: Molecular Medicine Reports Volume: in press
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Phosphorylation status of human RNA-binding protein 8A in cells and its inhibitory regulation by Magoh (2014)
  • Author(s): Y.Ishigaki, Y.Nakamura, T.Tatsuno, S.Ma, N.Tomosugi
  • Journal Title: Exp. Biol. Med. (Maywood) Volume: 240 Issue: 4 Pages: 438-445
  • DOI: 10.1177/1535370214556945
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] RNA binding protein RBM8A (Y14) and MAGOH localize to centrosome in human A549. (2014)
  • Author(s): Ishigaki Y, Nakamura Y, Tatsuno T, Hashimoto M, Iwabuchi K, Tomosugi N.
  • Journal Title: Histochem Cell Biol Volume: 141 Issue: 1 Pages: 101-109
  • DOI: 10.1007/s00418-013-1135-4
  • Peer Reviewed
• [Journal Article] Depletion of RNA-binding protein RBM8A (Y14) causes cell cycle deficiency and apoptosis in human cells. (2013)
  • Author(s): Ishigaki Y, Nakamura Y, Tatsuno T, Hashimoto M, Shimasaki T, Iwabuchi K, Tomosugi N.
  • Journal Title: Exp Biol Med Volume: 238 Issue: 8 Pages: 889-897
  • DOI: 10.1177/1535370213494646
  • Peer Reviewed
• [Presentation] 中心体に局在するExon Junction Complex (EJC)因子の解析 (2015)
  • Author(s): 辰野 貴則、中村 有香、石垣 靖人
  • Organizer: 第３８回日本分子生物学会
  • Place of Presentation: 神戸ポートアイランド（兵庫県神戸市）
  • Year and Date: 2015-12-01
• [Presentation] Localization of RNA-binding complex EJC on centrosome (2015)
  • Author(s): Yasuhito Ishigaki, Takanori Tatsuno, Yuka Nakamura
  • Organizer: 第６７回日本細胞生物学会大会
  • Place of Presentation: タワーホール船堀（東京都江戸川区）
  • Year and Date: 2015-06-30
• [Presentation] 中心体局在にするmRNA結合因子 (2014)
  • Author(s): 辰野貴則, 中村有香, 石垣靖人
  • Organizer: 第３７回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] Role of RNA binding protein in cell cycle progression and centrosome maturation in human tumor cells (2014)
  • Author(s): 石垣靖人, 中村有香, 辰野貴則, 橋本光正, 島崎猛夫, 岩淵邦芳, 友杉直久
  • Organizer: 19th World Congress on Advances in Oncology & 17th International Symposium of Molecular Medicine
  • Place of Presentation: アテネ（ギリシャ）
  • Year and Date: 2014-10-09 – 2014-10-11
• [Presentation] 細胞内mRNA-タンパク質構造の可視化 (2013)
  • Author(s): 石垣靖人, 中村有香, 辰野貴則, 島田ひろき, 八田稔久, 桑畑 進, 中川秀昭, 竹上 勉, 友杉直久
  • Organizer: 日本顕微鏡学会 第69回学術講演会
  • Place of Presentation: 大阪府吹田市 ホテル阪急エキスポパーク
• [Presentation] 中心体局在因子RBM8Aのリン酸化制御の解析 (2013)
  • Author(s): 石垣靖人, 中村有香, 辰野貴則, 馬 少幅, 中川秀昭, 竹上 勉, 友杉直久
  • Organizer: 第65回日本細胞生物学会大会
  • Place of Presentation: 愛知県名古屋市 ウインクあいち
• [Remarks] 金沢医科大学総合医学研究所
  • URL: http://www.kanazawa-med.ac.jp/mri/research.html