細胞運動における細胞骨格の動的性質

1990 Fiscal Year Annual Research Report

• Project/Area Number: 01304059
• Research Institution: Ochanomizu University
• Principal Investigator: 能村 堆子 お茶の水女子大学, 理学部, 教授 (30022578)
• Co-Investigator(Kenkyū-buntansha): 久永 眞市 東京工業大学, 生命理工学部, 助教授 (20181092) ; 高橋 景一 東京大学, 理学部, 教授 (40011481) ; 浜口 幸久 東京工業大学, 理学部, 教授 (70016161) ; 月田 承一郎 岡崎国立共同研究機構, 生理学研究所, 教授 (50155347) ; 馬渕 一誠 東京大学, 教養学部, 助教授 (40012520)
• Keywords: 細胞骨格 / アクチンーミオシン / ダイニンーチュ-ブリン / 動的不安定性 / 再構成運動系 / ニュ-ロフイラメント / 分裂装置 / 微小管

Research Abstract

生物体における骨格は、支持構造としての不変性と,生体の成長に伴う可変性をあわせもつことが要求される。この2つの相反した機能は、生体の構成単位である細胞にも要請されている。主にアクチンーミオシン系とチュ-ブリンーダイニン系から構成されている細胞骨格は細胞内で恒常的にあるいは細胞周期に同調して、骨格たる重合体と可変性を担う単量体との間に、量的変動をくり返しながら、要請された機能発現を充足している。こゝでは生成するばかりでなく、消減することもまた重要な機能の一つである。
本研究はこれらの系の動的性質がいかに細胞の機能と密接に関連しているかを、細胞運動を中心とした個々の系の具体例について,各々の研究者が計画に従って解析を行った。
(1)微小管の動的不安定性がMaps,dyneinの結合によっていかに制御されているかを解析し、dyneinの効果は、Mapsー微小管とPCー微小管で異なることを,Map2では結合siteが脱重合の停止重合の開始点であることを確定した。
(2)包転分子モ-タ-の解析。dyneinとmyosinについて、再構成運動系を用い、又クレオタイド特異性,ATP加水分解速度と運動の速さとの相関は高くないことを知った。平滑筋ミオシン,車軸築ミオシンを用いた運動の極性,アクチン分子内切断標品の運動速度について興味ある結果を得た。
(3)細胞内で機能分化した微小管の構成について,ウニ卵分裂装置で調べた結果,紡錘体,甲状体の微小管は異なるアイソタイプチュ-ブリンから構成されていることがわかった。
(4)ニュ-ロフイラメントの機能とリン酸化についてはHサブユニットの尾部のリン酸化は微小管との相互作用に,トリプレット蛋白の頭部は繊維の断片化に関与していることが明らかとなった。

Research Products

• [Publications] Tanaka.M.& MikiーNoumura.T.: "Stepwise sliding disintegration of <Tetrahymena>___ー ciliary axoーnemes at higher concentrations of ATP" Cell Motility & the Cytoskelton. 9. 191-204 (1988)
• [Publications] Yamao,Y.& MikiーNoumura,T.: "Effects of hexyleneglycol on meiotic division of starfish oocytes" Zool.Sci.5. 563-572 (1988)
• [Publications] Yamada,N.& MikiーNoumura,T.: "Isolated centralーpair microtubules from <Chlamydomonas>___ー <reinーhardtii>___ー" 6. 1088 (1989)
• [Publications] Nagata,A.& MikiーNoumura,T.: "Aster formation in the activated sea urchin eggs" Zool.Sci.6. 1152 (1989)
• [Publications] 三木(能村) 堆子: "卵細胞微小管の機能" 蛋白質、核酸、酵素. 34. 1626-1637 (1989)
• [Publications] Y.Mimori & T.MikiーNoumura: "SLIDING MOVEMENT OF MICROTUBULES FROM THE BUNDLE OF DYNEIN MICROTUBULES COMPLEX" Zool.Sci.(1990)
• [Publications] M.Mori & T.MikiーNoumura: "CIRCULAR SLIDING MOVEMENT <IN>___ー <VITRO>___ー OF DOUBLET MICROTUBULES INDUCED BY DYNEINS OF <TETRAHYMENA>___ー CILIA" Zool.Sci.(1990)
• [Publications] T.MikiーNoumura: "Darkーfield microscopic study of microtubules in solution" International Review of Cytology. 122. 65-104 (1990)
• [Publications] Hotani,H.and T.Horio.: "“Dynamics of Microtubules visualized by Darkーfield Microscopy:Treadmilling and Dynamic Instability."" Cell Motil.Cytskl.10. 229-236 (1988)
• [Publications] Vale,R.D.and H.Hotani.: "“Formation of Membrane Networks in vitro by Kinesinーdrive Microtubule Movement."" J.Cell Biol.107. 2233-2241 (1988)
• [Publications] Mandelkow,E.,E.ーM.Mandelkow,H.Hotani,B.Hess and S.C.Muller.: "“Spatial Pattern from Oscillating Microtubules."" Science.246. 1291-1293 (1989)
• [Publications] Hotani,H.and H.Miyamoto.: "“Dynamic Features of Microtubules as Visualized by Darkーfield Microscopy."" Advanc.Biophys. 26. 135-156 (1990)
• [Publications] Toyoshima,Y.Y.,Kron,S.J.& Spudich,J.A.: "The myosin step size:Measurement of the unit displacement per ATP hydrolysed in an in vitro assay." Proc.Natl.Acad.Sci.87. 7130-7134 (1990)
• [Publications] Schwyter,D.H.,Kron,S.J.,Toyoshima,Y.Y.,Spudich,J.A.& Reisler,E.: "Subtilisin cleavage of actin inhibits in vitro sliding movement of actin filaments over myosin." J.Cell Biol.111. 465-470 (1990)
• [Publications] Toyoshima,Y.Y.: "Bidirectional movement of actin filaments along tracks of heavy meromyosin and native thick filaments." J.Cell Sci.(1991)
• [Publications] Shimizu,T.,Furusawa,K.,Ohashi S.,Toyoshima,Y.Y.,Okuno,M.,Malik,F.& Vale R.D.: "Nucleotide specificity of the enzymatic and motile activities of dynein,kinesin and heavy meromyosin." J.Cell Biol.(1991)
• [Publications] Yamada,A.,N.Ishii,T.Shimmen and K.Takahashi: "MgーATP activity and motility of native thick filaments isolated from the anterior byssus retractor muscle of Mytilus edulis." J.Muscle Res.Cell Motility. 10. 124-134 (1989)
• [Publications] Abe,O.,T.Yamada,K.Takahashi and H.Sugi: "Initiation of active force development in a molluscan smooth muscle by laser photolysis of caged ATP." Proc.Japan Academy Ser.B. 65. 165-168 (1989)
• [Publications] Katada,J.,C.Shingyoji and K.Takahashi: "Effects of detergents used to demembranate the seaーurchin spermatozoa on the reactivated flagellar movement,with special reference to the rotatability of the plane of flagellar beat." Cell Struct.Funct.14. 751-758 (1989)
• [Publications] Yoshimura,K.and K.Takahashi: "Partial extraction of inner dynein arms from an outer armーless mutant of Chlamydomonas reinhadtii." Cell Struct.Funct.14. 741-749 (1989)
• [Publications] Eshel,D.,C.Shingyoji,K.Yoshimura,B.H.Gibbons,I.R.Gibbons and K.Takahashi: "Transient behavior of sea urchin sperm flagella following an abrupt change in beat frequency." J.exp.Biol.15. 441-451 (1990)
• [Publications] Yamada,A.,N.Ishii and K.Takahashi: "Direction and speed of actin filaments moving along thick filaments isolated from molluscan smooth muscle." J.Biochem.108. 341-343 (1990)
• [Publications] Shingyoji,C.,I.R.Gibbons,A.Murakami and K.Takahashi: "Effects of imposed head vibration on the stability and waveform of flagellar beating in sea urchin spermatozoa." J.exp.Biol.,in press.(1991)
• [Publications] Shingyoji,C.,J.Katada,K.Takahashi and I.R.Gibbons: "Rotating the plane of imposed vibration can rotate the plane of flagellar beating in sea urchin sperm without twisting the axoneme." J.Cell Sci.,in press.(1991)
• [Publications] Takahashi,K.,C.Shingyoji,J.Katada,D.Eshel and I.R.Gibbons: "Polarity in spontaneous unwinding after prior rotation of the flagellar beat plane in sea urchin spermatozoa." J.Cell Sci.,in press.(1991)
• [Publications] Ishii,N.,F.Mitsumori and K.Takahashi: "Changes in sarcoplasmic metabolite concentrations and pH associated with the catch contraction and relaxation of the anterior byssus retractor muscle of Mytilus edulis measured by phosphorusー31 nuclear magnetic resonance." J.Muscle Res.Cell Motil.,12,in press.
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• [Publications] HigashiーFujime,S.: "Actinーinduced elongation of fibers composed of cytoplasmic membrane from Nitella." Protoplasma. Suppl.2. 27-36 (1988)
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• [Publications] Hisanaga,S.,Inagaki,M.& Hirokawa,N.: "Phosphorylation of the native and the reassembled neurofilaments composed of triplet proteins by the catalytic subunit of the cAMPーdependent protein kinase." In preparation.
• [Publications] Oka,M.T.,Arai,T.,& Hamaguchi,Y.: "Heterogeneity of microtubules in dividing sea urchin eggs revealed by immunofluorescence microscopy: Spindle microtubules are composed of tubulin isotypes different from those of astral microtubules." Cell Motil.Cytoskeleton. 16. 239-250 (1990)
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• [Publications] 長谷川 成人、新井 孝夫、井原 康夫: "Immunochemical evidence that fragments of phosphorylated MAP5 (MAP1B) are bound to neurofibrillary tangles in Alzheimer's disease." Neuron. 4. 909-918 (1990)
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• [Publications] 岡 美佳子、新井 孝夫、浜口 幸久: "Microinjection of the monoclonal antiーtubulin antibody YL1/2 inhibits cleavage of sand dollar eggs." Cell Struct.Funct.15. 37-80 (1990)
• [Publications] Takaaki Kobayashi: "ISOLATION OF MORPHOLOGICAL MUTANTS FROM C6 GLIOMA CLONAL CELLS AND CHARACTERIZATION OF LAMININ DISTRIBUTION IN THEM." Cell Struc.Func.15. 493 (1990)
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• [Publications] Aihara,M.,Kasukawa,H.,Fujii,R.and Oshima,N.: "Chemical characterization of reflecting platelets in motile iridophores of blue damselfish." Comp.Biochem.Physiol.92B. 533-536 (1989)
• [Publications] Fujii,R.,Kasukawa,H.,Miyaji,K.and Oshima,N.: "Mechanism of skin coloration and its changes in blueーgreen damselfish,<Chromis>___ー <viridis.>___ー" Zool.Sci.477-486 (1989)
• [Publications] Nagaishi,H.,Nishi,H.,Fujii,R.and Oshima,N.: "Correlation between body color and behavior in the upsideーdown catfish,<Synodontis>___ー <nigriventris.>___ー" Comp.Biochem.Physiol.92A. 323-326 (1989)
• [Publications] Oshima,N.Kasukawa,H.and Fujii,R.: "Control of chromatophore movements in blueーgreen damselfish,<Chromis>___ー <viridis>___ー." Comp.Biochem.Physiol.93C. 239-245 (1989)
• [Publications] Nagaishi,H.Oshima,N.and Fujii,R.: "Lightーrefrecting properties of the iridophores of the neon tetra,<Paracheirodon>___ー <innesi.>___ー" Comp.Biochem.Physiol.95A. 337-341 (1990)
• [Publications] Fujii,R.,Wakatabi,H.and Oshima,N.: "Inositol 1,4,5ーtrisphosphate signals the motile response of fish chromatophoresーーーI.Aggregation of pigment in the tilapia melanoーphore." J.Exp.Zool.(1991)
• [Publications] Fujii,R.,Hayashi,H.,Toyohara,J.and Nishi,H.: "Analysis of the reflection of light from motile iridophores of the dark sleeper,<Odontobutis obscura obscura>___ー." Zool.Sci.(1991)
• [Publications] Mogami,Y.,Pernberg,J.& Machemer,H.: "Messenger role of calcium in ciliary electromotor coupling:a reassessment." Cell Calcium. 11. 665-673 (1990)
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• [Publications] Mogami,Y.,Pernberg,J.& Machemer,H.: "Five different states of ciliary activity in the epaulette of echinoplutei." Journal of Experimental Biology. 155. (1991)
• [Publications] Mogami,Y.,Fujima,K.& Baba,S.A.: "Ciliary beating in three dimensions: step of a quantitative description." Journal of Mathematical Biology. (1991)
• [Publications] 宝谷 紘一、伊藤 知彦: "微小管の動的性質(第11章) 細胞増殖・細胞運動 (シリ-ズ分子生物学の進歩・第7巻)" 丸善(株), 251 (1989)
• [Publications] Kron,S.J.,Toyoshima,Y.Y.,Uyeda,T.Q.P.& Spudich,J.A.: "Assays for actin sliding movement over myosin coated surfaces.In“Methods in Enzymology"" Academic Press, (1990)
• [Publications] Oka,M.T.,Arai,T.,& Hamaguchi,Y.: "Heterogeneity and assemblyーdisassembly of microtubules in dividing sea urchin eggs." In Proc.7th Internat.Echinoderm Conference,A.A.Balkema Pub.Rotterdam, (1990)
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• [Publications] 藤井 良三: "第12章 色素細胞と体色変化.メダカの生物学(江上信雄他編) 185ー199頁," 東大出版会,東京., 315 (1990)
• [Publications] 藤井 良三・大島 範子編: "生物学" 八千代出版,東京., 317 (1990)