Molecular function(s) of MOT48 in the dynein pre-assembly

• Project/Area Number: 17K15117
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Cell biology
• Research Institution: Osaka University
• Principal Investigator: Yamamoto Ryosuke 大阪大学, 理学研究科, 助教 (10743114)
• Project Period (FY): 2017-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2018: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2017: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 繊毛 / 前集合 / pre-assembly / MOT48 / TWI1 / PF13 / ダイニン / Chlamydomonas / クラミドモナス / 運動性 / 繊毛ダイニン / ida10-2 / PIH1 / 運動性繊毛 / ida10 / 前集合 (pre-assembly) / 鞭毛

Outline of Final Research Achievements

Motility of cilia is driven by the gigantic motor-protein complexes called "ciliary dyneins". Ciliary dyneins are folded/assembled from their subunits (HC/IC/LC) in the cytoplasm, and this process is called "pre-assembly". Defects in the dynein pre-assembly cause various severe human diseases, but the underlying molecular mechanism(s) of the dynein pre-assembly has been poorly understood. In this study, by analyzing several pre-assembly deficient mutants of a green alga (Chlamydomonas reinhardtii) we aimed to elucidate the molecular mechanism(s) of the dynein pre-assembly. We identified ciliary dynein species, which were folded/assembled by the three Chlamydomonas pre-assembly factors (PF13/MOT48/TWI1). Further, we found that the functions of these three dynein pre-assembly factors were somewhat redundant and complementary, suggesting the mechanism of dynein pre-assembly is far more complex than has been previously thought.

Academic Significance and Societal Importance of the Research Achievements

繊毛ダイニンの細胞質内における組み立て/折り畳み(前集合)異常は、左右軸逆位/気管支炎/不妊等を含む重篤な人体疾患の引き金となる。しかしながら、その重要性にも関わらず、繊毛ダイニン前集合機構の分子基盤には不明な点が多い。我々は、2本の運動性繊毛を持つ単細胞緑藻クラミドモナスの変異株を用いて、遺伝学的/生化学的/構造生物学的な手法により前集合の分子基盤の一端の解明に努めた。その結果、2種の新規前集合異常株(mot48-2及びtwi1株)の単離に成功し、複数種の繊毛ダイニン前集合因子(PF13/MOT48/TWI1)が具体的にどの繊毛ダイニン種の組み立て/折り畳みに関与するのかを明らかにした。

Research Products

• [Int'l Joint Research] エモリー大学/コネチカット大学 健康センター/SUNY アップステート・メディカル大学(米国)
• [Int'l Joint Research] ポールシェラー研究所(スイス)
• [Int'l Joint Research] University of Connecticut Health Center(米国)
• [Int'l Joint Research] Paul Scherrer Institute(スイス)
• [Int'l Joint Research] ポール・シェラー研究所(スイス)
• [Int'l Joint Research] コネチカット大学医学センター(米国)
• [Int'l Joint Research] ポール・シェラー研究所(スイス)
• [Journal Article] A dynein-associated photoreceptor protein prevents ciliary acclimation to blue light (2021)
  • Author(s): Kutomi O, Yamamoto R, Hirose K, Mizuno K, Nakagiri Y, , , , , , , Shibata D, Shibata M, Shiba K, Kita M, Kigoshi H, Tanaka Y, Yamasaki Y, Asahina Y, Song C, Nomura M, Nomura M, Nakajima A, Nakachi M, Yamada L, Nakazawa S, Sawada H, Murata K, Mitsuoka K, Ishikawa T, Wakabayashi K, Kon T, Inaba K
  • Journal Title: Science Advances Volume: 7 Issue: 9
  • DOI: 10.1126/sciadv.abf3621
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Mutations in PIH proteins MOT48, TWI1 and PF13 define common and unique steps for preassembly of each, different ciliary dynein (2020)
  • Author(s): Ryosuke Yamamoto, Shiho Yanagi, Masahito Nagao, Yuya Yamasaki, Yui Tanaka, Winfield S. Sale, Toshiki Yagi, Takahide Kon
  • Journal Title: PLOS Genetics Volume: 16 Issue: 11 Pages: e1009126-e1009126
  • DOI: 10.1371/journal.pgen.1009126
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Transport toward the center of the cell: structure & mechanism of microtubule-based systems [MOT7/DYBLUP, a novel dynein subunit with a BLUF domain]. (2021)
  • Author(s): Takahide Kon, Hiroshi Imai, Ryosuke Yamamoto
  • Organizer: 台湾清華大学‐大阪大学 国際シンポジュウム (2021 NTHU-OU Virtual Symposium on Biological and Medical Sciences)
  • Int'l Joint Research
• [Presentation] クライオ電子線トモグラフィー法による光感受性ドメインを持つ新規ダイニン軽鎖局在決定 (2020)
  • Author(s): 山本遼介, 中桐侑平, 久冨理, 今井洋, Song Chihong, 村田和義, 光岡薫, 若林憲一, 石川尚, 稲葉一男, 昆隆英
  • Organizer: 日本顕微鏡学会 第76回学術講演会
• [Presentation] Structural/functional analyses on MOT7, a novel light chain of ciliary dynein f/I1. (2019)
  • Author(s): Ryosuke Yamamoto, Yuuhei Nakagiri, Osamu Kutomi, Hiroshi Imai, Chihong Song, Kazuyoshi Murata, Ken-ichi Wakabayashi, Takashi Ishikawa, Kazuo Inaba, Takahide Kon
  • Organizer: 第57回 日本生物物理学会 年会
• [Presentation] Cryo-electron tomography revealed localization of a novel ciliary dynein subunit, MOT7. (2019)
  • Author(s): R. Yamamoto, Y. Nakagiri, O. Kutomi, H. Imai, C. Song, K. Murata, K. Mitsuoka, K. Wakabayashi, T. Ishikawa, K. Inaba, T. Kon
  • Organizer: 生理研研究会
• [Remarks] 光で繊毛運動を調節する新規タンパク質の発見
  • URL: https://resou.osaka-u.ac.jp/ja/research/2021/20210302_2