1989 Fiscal Year Annual Research Report

真核細胞における鞭毛運動の分子機構の解明

Research Project

Project/Area Number	01304006
Research Institution	The University of Tokyo
Principal Investigator	森沢正明東京大学, 理学部, 教授 (40013594)
Co-Investigator(Kenkyū-buntansha)	中村健一広島女子大学, 家政学部, 教授 (50094694) 馬場昭次お茶の水女子大学, 理学部, 助教授 (80011691) 林博司名古屋大学, 理学部, 助教授 (00022608) 高橋景一東京大学, 理学部, 教授 (40011481) 毛利秀雄東京大学, 教養学部, 教授 (70012268)
Keywords	鞭毛繊毛運動 / 滑り運動 / チュ-ブリン / ダイニン / カルシウム / リン酸化反応 / コンホメ-ション変化 / 突然変異
Research Abstract	本総合研究初年度において,まず鞭毛繊毛運動を担う主要タンパク質であるtubulin,dyneinの相互作用に関する研究の新しい展望が開かれた。即ち,tubulin-オ-バ-レイ法が開発され,数種のtubulin結合タンパクが同定された。21sdyneinについては,そのB鎖のコンホメ-ション変化がATP分解機能に重要である事が示された。また外腕以外のdyneinが微小管を回転させることや,回転運動のモ-タ-が鞭毛内に存在することが新に発見されたこと,更には鞭毛が微小な高速運動をするという発見がnoture誌に取り上げられたことは特筆に価する。次に鞭毛運動のCa^<2+>,CAMDなどによる調節機構について,細胞内Ca^<2+>濃度画像解析システム,TPP電極による微細細胞の膜電位測定法,イオントフォレシス法等様々な生化学的手法が駆使され,特定の微小管間で能動的滑りが起こり易いこと,精子運動開始は鞭毛基動へのCa^<2+>の流入が引き金になること及びその際に膜電位が発生することなどが新に見い出された。更に運動調節に関与するリン酸化タンパク質の大量精製と抗体の作成が行なわれ,将来蛍光抗体法等による運動調節部位の特定などの道を拓いた。カルモジュリンとは異なる新しいCa^<2+>給合タンパク質の発見と一次構造の決定,鞭毛の波形と遊泳速度の調節因子の精製は今後の鞭毛運動調節機構の生化学的側面からの研究に新な展望を開く研究として期待される。繊毛運動の調節機構に関しては,ウニ幼生の繊毛運動がド-バミン,セロトニンで調節されている事が見い出され,ゾウリムシの繊毛の性質は一様ではなく,CAMD感受性が異なる繊毛が分布している事が示され,更に突然変異株を用いた研究から,Ca^<2+>の細胞内流入が繊毛打頻度を調節していることが示された。植物性鞭毛虫類クラミドモナスでは,dynein内腕欠失変異株が多数単離され,解析の結果,それ等が4種に分類できることが示された。またそれ等の変異の遺伝子地図上の位置が決定された。

Research Products

(32 results)

All Other

All Publications (32 results)

[Publications] Morisawa M.: "Regulation of sperm motility by osmotiy pressure" In"Mechanism of Fertilization:Plants to Human"B.Dale ed.NATO ASI series on cell Biology,Springer-Verlag,Berlin Heiderberh,New York.
[Publications] Morisawa M.: "Acquisition and initiation of sperm motility" In"Control of Sperm Motility"C.Gagnon ed.CRC Press,Inc.Boca Raton.
[Publications] Okuno M.: "Effects of calcium on the motility of rainbow trout sperm flagella demembranated with Triton X-100" Cell Motility and Cytoskeleton. 14. 194-200 (1989)
[Publications] Inaba K.: "dynamic conformational changes of 21 s dynein ATPase coupled with ATP hydrolysis revealed by proteolytic digestion" The Journal of Biological Chemistry. 264. 8384-8388 (1989)
[Publications] Inaba K.: "Two states of the comformation of 21 s outer arm dynein coupled with ATP hydrolysis" The Journal of Biological Chemistry(TokYo). 106. 349-354 (1989)
[Publications] Inaba K.: "Comformational changes of the chain of the outer arm dynein from sea urchin sperm flagella coupled with ATP hydrolysis" The Journal of Biological Chemistry.
[Publications] Inaba K.: "Anthraniloyl ATP,a fluorescent analog of ATp,as a substrate for dynein ATpase and flagellar motility" Archives of Biochemistry and Biophysics. 274. 209-215 (1989)
[Publications] Yoshimura K.: "Partial extraction of inner dynein arms from an outer arm-less mutant of Chlamydomonas reinbardtii" Cell Structure and Function. 14. 741-749 (1989)
[Publications] Katada J.: "Effects of detergents used to demembranate the sea-urchin spermatozoa on teh reactivated flagellar movement.with special reference to the totatability of the plane fo flagellar beat" Cell Structure and Function. 14. 751-758 (1989)
[Publications] Abe O: "Initiation of ctie force development in a molluscan smooth muscle by laser photolysis of caged ATP" proceedings of Japan Academy,Ser B. 65. 165-168 (1989)
[Publications] Eshel D.: "Transient behaviow of sea urchin sperm flagella following" Journal of Experimental Biology. in press.
[Publications] Hayashi H.: "Measurement of ATPase activity of immobilized myosin heads" Journal of Biochemistry(Tokyo). 105. 875-877 (1989)
[Publications] Kamimura S.: "High-frequency nanometre-scale vibration in 'quiescent'flagellar axoneme" Nature. 340. 476-478 (1989)
[Publications] Baba S.: "Discrete nature of flagellar bending detected by digital image analysis." Biomathematics. in press.
[Publications] Nakamura K.: "Characterization of an axonemal protein from Tetrahymena cilia" Bull.Hiroshima Women s University. 25. 21-28 (1989)
[Publications] Nakaoka Y.: "Localization of photoreceptor in Paramecium bursaria" journal of Comparative Physiology A. 165. 637-643 (1989)
[Publications] Nakaoka Y.: "Effects of cyclic nucleotides and intracellular Ca^<2+> on voltage activatid ciliary beating in paramecium" journal of Comparative Physiology A. in press.
[Publications] Oka T.: "Inactivation and activation of inward current during adaptation to potassium ions in Paramecium caudatum" Cell Structure and Function. 14. 209-216 (1989)
[Publications] Nakajima K.: "Circadian changes of photosensitivity in Paramecium bursaria" Journal of Experimental Biology. 144. 43-51 (1989)
[Publications] Sakakibara H.: "Functional recombinaton of outer dynein arms with outer arm-missing flagellar axonemes of a Chlamydomonas mutant" Journal of Cell Science. 92. 77-83 (1989)
[Publications] Kamiya R.: "Strategies for the selection of Chlamydomonas dynein mutant" Methods in Enzymology. in press.
[Publications] Kagami O.: "Strikingly low ATPase in flagellar axonemes of a Chlamidomonas mutant missing outer dynein arms" European Journal of Biochemistry. in press.
[Publications] 神谷律: "鞭毛運動とダイニン内腕・外腕" 生物物理. 29. 231-236 (1989)
[Publications] Vale D.: "In vitro motility assays for kinesin and dynein" In"Cell Movement"(Alan R,Liss,Inc.). 2. 287-294 (1989)
[Publications] Vale R.: "Microtubule translocation properties of intact and proteolytically digested dyneins from Tetrahymena cilia" Journal of Cell Biology. 108. 2327-2334 (1989)
[Publications] Toyoshima Y.: "Bidirectional movement of actin filaments along tracks of myosin heads" Nature. 341. 154-156 (1989)
[Publications] Kron S.J.: "Assays for actin sliding movement over myoshin coated surfaces" Methods in Enzymology. in press.
[Publications] 豊島陽子: "アクチンシオミンの運動再構成系" 蛋白質核酸酵素. 34. 1480-1486 (1989)
[Publications] 稲葉一男・毛利秀雄: "鞭毛・繊毛運動の分子機構蛋白質、核酸酵素「細胞骨格の機能」(酒井彦一編)" 共立出版, 1989
[Publications] 上村慎治: "生物理実験技術『光学顕微鏡によるナノメ-タ-精度での運動解析』" 吉岡書店, 10 (1990)
[Publications] 上村慎治: "細胞・細胞運動の定量法" ニュ-サイエンス社, 10 (1990)
[Publications] 上村慎治: "アトバンテストテクノロジ-生物工学編第13巻生体物理工学第3章ナノメ-トル精度での顕微計測、顕微操作" 丸善株式会社出版事業部, 20 (1990)

1989 Fiscal Year Annual Research Report

真核細胞における鞭毛運動の分子機構の解明

Principal Investigator

森沢 正明 東京大学, 理学部, 教授 (40013594)

Research Products

[Publications] Morisawa M.: "Regulation of sperm motility by osmotiy pressure" In"Mechanism of Fertilization:Plants to Human"B.Dale ed.NATO ASI series on cell Biology,Springer-Verlag,Berlin Heiderberh,New York.

[Publications] Morisawa M.: "Acquisition and initiation of sperm motility" In"Control of Sperm Motility"C.Gagnon ed.CRC Press,Inc.Boca Raton.

[Publications] Okuno M.: "Effects of calcium on the motility of rainbow trout sperm flagella demembranated with Triton X-100" Cell Motility and Cytoskeleton. 14. 194-200 (1989)

[Publications] Inaba K.: "dynamic conformational changes of 21 s dynein ATPase coupled with ATP hydrolysis revealed by proteolytic digestion" The Journal of Biological Chemistry. 264. 8384-8388 (1989)

[Publications] Inaba K.: "Two states of the comformation of 21 s outer arm dynein coupled with ATP hydrolysis" The Journal of Biological Chemistry(TokYo). 106. 349-354 (1989)

[Publications] Inaba K.: "Comformational changes of the chain of the outer arm dynein from sea urchin sperm flagella coupled with ATP hydrolysis" The Journal of Biological Chemistry.

[Publications] Inaba K.: "Anthraniloyl ATP,a fluorescent analog of ATp,as a substrate for dynein ATpase and flagellar motility" Archives of Biochemistry and Biophysics. 274. 209-215 (1989)

[Publications] Yoshimura K.: "Partial extraction of inner dynein arms from an outer arm-less mutant of Chlamydomonas reinbardtii" Cell Structure and Function. 14. 741-749 (1989)

[Publications] Katada J.: "Effects of detergents used to demembranate the sea-urchin spermatozoa on teh reactivated flagellar movement.with special reference to the totatability of the plane fo flagellar beat" Cell Structure and Function. 14. 751-758 (1989)

[Publications] Abe O: "Initiation of ctie force development in a molluscan smooth muscle by laser photolysis of caged ATP" proceedings of Japan Academy,Ser B. 65. 165-168 (1989)

[Publications] Eshel D.: "Transient behaviow of sea urchin sperm flagella following" Journal of Experimental Biology. in press.

[Publications] Hayashi H.: "Measurement of ATPase activity of immobilized myosin heads" Journal of Biochemistry(Tokyo). 105. 875-877 (1989)

[Publications] Kamimura S.: "High-frequency nanometre-scale vibration in 'quiescent'flagellar axoneme" Nature. 340. 476-478 (1989)

[Publications] Baba S.: "Discrete nature of flagellar bending detected by digital image analysis." Biomathematics. in press.

[Publications] Nakamura K.: "Characterization of an axonemal protein from Tetrahymena cilia" Bull.Hiroshima Women s University. 25. 21-28 (1989)

[Publications] Nakaoka Y.: "Localization of photoreceptor in Paramecium bursaria" journal of Comparative Physiology A. 165. 637-643 (1989)

[Publications] Nakaoka Y.: "Effects of cyclic nucleotides and intracellular Ca^<2+> on voltage activatid ciliary beating in paramecium" journal of Comparative Physiology A. in press.

[Publications] Oka T.: "Inactivation and activation of inward current during adaptation to potassium ions in Paramecium caudatum" Cell Structure and Function. 14. 209-216 (1989)

[Publications] Nakajima K.: "Circadian changes of photosensitivity in Paramecium bursaria" Journal of Experimental Biology. 144. 43-51 (1989)

[Publications] Sakakibara H.: "Functional recombinaton of outer dynein arms with outer arm-missing flagellar axonemes of a Chlamydomonas mutant" Journal of Cell Science. 92. 77-83 (1989)

[Publications] Kamiya R.: "Strategies for the selection of Chlamydomonas dynein mutant" Methods in Enzymology. in press.

[Publications] Kagami O.: "Strikingly low ATPase in flagellar axonemes of a Chlamidomonas mutant missing outer dynein arms" European Journal of Biochemistry. in press.

[Publications] 神谷律: "鞭毛運動とダイニン内腕・外腕" 生物物理. 29. 231-236 (1989)

[Publications] Vale D.: "In vitro motility assays for kinesin and dynein" In"Cell Movement"(Alan R,Liss,Inc.). 2. 287-294 (1989)

[Publications] Vale R.: "Microtubule translocation properties of intact and proteolytically digested dyneins from Tetrahymena cilia" Journal of Cell Biology. 108. 2327-2334 (1989)

[Publications] Toyoshima Y.: "Bidirectional movement of actin filaments along tracks of myosin heads" Nature. 341. 154-156 (1989)

[Publications] Kron S.J.: "Assays for actin sliding movement over myoshin coated surfaces" Methods in Enzymology. in press.

[Publications] 豊島陽子: "アクチンシオミンの運動再構成系" 蛋白質核酸酵素. 34. 1480-1486 (1989)

[Publications] 稲葉一男・毛利秀雄: "鞭毛・繊毛運動の分子機構蛋白質、核酸酵素「細胞骨格の機能」(酒井彦一編)" 共立出版, 1989

[Publications] 上村慎治: "生物理実験技術『光学顕微鏡によるナノメ-タ-精度での運動解析』" 吉岡書店, 10 (1990)

[Publications] 上村慎治: "細胞・細胞運動の定量法" ニュ-サイエンス社, 10 (1990)

[Publications] 上村慎治: "アトバンテストテクノロジ-生物工学編第13巻生体物理工学 第3章ナノメ-トル精度での顕微計測、顕微操作" 丸善株式会社出版事業部, 20 (1990)

森沢正明東京大学, 理学部, 教授 (40013594)

[Publications] 上村慎治: "アトバンテストテクノロジ-生物工学編第13巻生体物理工学第3章ナノメ-トル精度での顕微計測、顕微操作" 丸善株式会社出版事業部, 20 (1990)