2000 Fiscal Year Annual Research Report

新たな生物分子モーターの探索

Research Project

Project/Area Number	09279102
Research Institution	The University of Tokyo
Principal Investigator	神谷律東京大学, 大学院・理学系研究科, 教授 (10124314)
Co-Investigator(Kenkyū-buntansha)	山本啓一千葉大学, 理学部, 教授 (70053361) 新免輝男姫路工業大学, 理学部, 教授 (80114510) 児玉孝雄九州工業大学, 情報工学部, 教授 (30034200) 豊島陽子東京大学, 大学院・総合文化研究科, 助教授 (40158043)
Keywords	ミオシン / ダイニン / 微小管 / グルカン合成酵素 / ADP / 植物細胞 / 鞭毛・繊毛 / 原形質流動
Research Abstract	今年度も植物ミオシンと軸糸ダイニンを中心として,モーター蛋白質の多様な性質を探る研究を行った.まず,植物細胞において,筋肉ミオシンの阻害剤BDMの効果を解析した.その結果,通常のミオシンとは構造的に異なる植物ミオシンもこの阻害剤によって活性が強く阻害され,それに伴って原形質流動が停止することが見いだされた(新免).さらに,生物界で最も高速の原形質流動を発生することで知られる車軸藻ミオシンの研究では,昨年度に続いて分子内の異なる部域を改変して,生体内外の性状を検討する試みを行った.今年度は,新たに頭部と尾部を機能を持った状態で,培養細胞内で発現させることに成功したが,このミオシンが高速で運動する機構はまだ不明である(山本).ダイニンの研究では,まずテトラヒメナ繊毛の内腕ダイニンaの性質を調べ,このダイニンは、重鎖1本と軽鎖3本からなる単頭の分子構造をもつこと、および,10μM程度の低濃度のADPによりATPase活性と運動活性が増大することを明らかにした(豊島).さらに,クラミドモナスの鞭毛ダイニン外腕と微小管の結合に関与し,その運動活性を調節している外腕ドッキング複合体の構造と機能を詳細に検討した.その結果,この複合体は単独で微小管上に24nm周期で結合できることが明らかになった.ダイニンが微小管上に規則正しく結合する機構として,重要であろう(神谷).また,モーター蛋白質以外の酵素系における運動性発現の可能性を調べる研究として,グルカン合成酵素GTFと基質の一つであるデキストランとの相互作用に注目し,活性部位を改変した酵素を作製した.改変酵素と基質の相互作用の解析から,GTFにはモーター蛋白質と共通した性質があると結論された.GTF一分子を蛍光観察することに成功しているので,近い将来,運動性の観察も可能であると期待される(児玉).

Research Products
(13 results)

All Other

All Publications (13 results)

[Publications] Wakabayashi,K.: "Transport and arrangement of the outer-dynein-arm docking complex in the flagella of Chlamydomonas mutants that lack outer dynein arms."Cell Motility and the Cytoskeleton. (印刷中).
[Publications] Kamiya,R.: "Analysis of cell vibration for assessing axonemal motility in Chlamydomonas"Methods. 22. 383-387 (2000)
[Publications] Isogai,N.: "Dominance between the two flagella during phototaxis in Chlamydomonas : dependence on the light/dark cycle caused by bodily rotation"Zoological Science. 17. 1261-1266 (2000)
[Publications] Yagi,T.: "Vigorous beating of Chlamydomonas axonemes lacking central pair/radial spoke structures in the presence of salts and organic compounds"Cell Motility and the Cytoskeleton. 46. 190-199 (2000)
[Publications] Yokota,E.: "Possible involvement of protein phosphorylation in the regulation of cytoplasmic streaming in root hair cells as revealed by a protein phosphatase inhibitor, calyculin A"Protoplasma. 211. 29-38 (2000)
[Publications] Tominaga,M.: "The role of plant villain in the organization of the actin cytoskeleton, cytoplasmic streaming and the artitecture of the transvacuolar strand in root hair cells of Hydrocharis"Planta. 210. 836-843 (2000)
[Publications] Shimmen,T.: "Plant myosins"Protoplasma. 214. 1-10 (2000)
[Publications] Igarashi,H.: "Isolation of a novel 190 kDa protein from tobacco BY-2 cells : possible involvement in the interaction between actin filaments and microtubules"Plant and Cell Physiology. 41. 920-931 (2000)
[Publications] Tominaga,M.: "Mechanism of inhibition of cytoplasmic streaming by a myosin inhibitor,2,3-butanedione monoxide"Protoplasma. 213. 46-54 (2000)
[Publications] Yokota,E.: "Actin cytoskeleton is responsible for the Change of cytoplasmic organization in root hair cells induced by a protein phosphatase inhibitor, calyculin A"Protoplasma. 213. 184-193 (2000)
[Publications] Kawakatsu,T.: "Interaction of actin filaments with the plasma membrane of Amoeba proteus : studies using a cell model and isolated plasma membrane"Cell Structure and Function. 25. 269-277 (2000)
[Publications] Kashiyama,T.: "Cloning and characterization of a myosin from Characean alga, the fastest motor protein in the world"J.Biochemistry. 127. 1065-1070 (2000)
[Publications] Hirakawa,E.: "Processive movement of single 22S dynein molecules occurs only at low ATP concentrations"Proc.Natl.Acad.Sci.USA. 97. 2533-2537 (2000)

2000 Fiscal Year Annual Research Report

新たな生物分子モーターの探索

Principal Investigator

神谷 律 東京大学, 大学院・理学系研究科, 教授 (10124314)

Research Products

[Publications] Wakabayashi,K.: "Transport and arrangement of the outer-dynein-arm docking complex in the flagella of Chlamydomonas mutants that lack outer dynein arms."Cell Motility and the Cytoskeleton. (印刷中).

[Publications] Kamiya,R.: "Analysis of cell vibration for assessing axonemal motility in Chlamydomonas"Methods. 22. 383-387 (2000)

[Publications] Isogai,N.: "Dominance between the two flagella during phototaxis in Chlamydomonas : dependence on the light/dark cycle caused by bodily rotation"Zoological Science. 17. 1261-1266 (2000)

[Publications] Yagi,T.: "Vigorous beating of Chlamydomonas axonemes lacking central pair/radial spoke structures in the presence of salts and organic compounds"Cell Motility and the Cytoskeleton. 46. 190-199 (2000)

[Publications] Yokota,E.: "Possible involvement of protein phosphorylation in the regulation of cytoplasmic streaming in root hair cells as revealed by a protein phosphatase inhibitor, calyculin A"Protoplasma. 211. 29-38 (2000)

[Publications] Tominaga,M.: "The role of plant villain in the organization of the actin cytoskeleton, cytoplasmic streaming and the artitecture of the transvacuolar strand in root hair cells of Hydrocharis"Planta. 210. 836-843 (2000)

[Publications] Shimmen,T.: "Plant myosins"Protoplasma. 214. 1-10 (2000)

[Publications] Igarashi,H.: "Isolation of a novel 190 kDa protein from tobacco BY-2 cells : possible involvement in the interaction between actin filaments and microtubules"Plant and Cell Physiology. 41. 920-931 (2000)

[Publications] Tominaga,M.: "Mechanism of inhibition of cytoplasmic streaming by a myosin inhibitor,2,3-butanedione monoxide"Protoplasma. 213. 46-54 (2000)

[Publications] Yokota,E.: "Actin cytoskeleton is responsible for the Change of cytoplasmic organization in root hair cells induced by a protein phosphatase inhibitor, calyculin A"Protoplasma. 213. 184-193 (2000)

[Publications] Kawakatsu,T.: "Interaction of actin filaments with the plasma membrane of Amoeba proteus : studies using a cell model and isolated plasma membrane"Cell Structure and Function. 25. 269-277 (2000)

[Publications] Kashiyama,T.: "Cloning and characterization of a myosin from Characean alga, the fastest motor protein in the world"J.Biochemistry. 127. 1065-1070 (2000)

[Publications] Hirakawa,E.: "Processive movement of single 22S dynein molecules occurs only at low ATP concentrations"Proc.Natl.Acad.Sci.USA. 97. 2533-2537 (2000)

神谷律東京大学, 大学院・理学系研究科, 教授 (10124314)