細胞運動における細胞骨格の動的性質

1989 年度 実績報告書

• 研究課題/領域番号: 01304059
• 研究機関: お茶の水女子大学
• 研究代表者: 能村 堆子 お茶の水女子大学, 理学部, 教授 (30022578)
• 研究分担者: 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) ; 馬渕 一誠 東京大学, 教養学部, 助教授 (40012520) ; 月田 承一郎 東京都臨床研, 室長 (50155347) ; 宝谷 紘一 帝京大学, 理工学部, 教授 (80025444)
• キーワード: 微小管 / in vitro再構成運動系 / Map2 / 動的不安定性 / ヤリイカの巨大神経 / 収縮環 / adherence junction / 鞭毛外腕欠如株

研究概要

微小管の動的不安定性のメカニズム
Map2やdyneinを結合させると微小管の動的不安定性が修飾されて、安定化に向かうことがわかった。Map2を微小管に結合させ、その分布を蛍光抗体を用いて可視化すると、微小管の脱重合相から重合相への移行がMap2の結合部位で起こることが明らかとなった。(宝谷、能村)
in vitro再構成運動系
(1)ウニ精子鞭毛から外腕を除去したのち、13S dyneinを抽出し、精製した。13S dyneinはon the slideで微小管を回転させるモ-タ-としての機能をもつ。(豊島)
(2)車軸藻ミオシンによる滑り速度は20um/sと非常に速い。この速度はCaによるリン酸化を通して制御されていることがわかった。(藤目)
(3)ブライトコントラスト位相差対物レンズの吸収板をかえて、高いコントラストの像がえられた。微小管の運動を解析精度を上げて、記録、追及できる。(上村)
in vivoでの解析
(1)クラミドモナスの鞭毛外腕欠如株から、KCLをもちて、内腕を部分抽出すると、鞭毛打頻度が減少することがわかった。(高橋)
(2)チラピアの培養黒色素胞の色素凝集反応が、inositoll,4,5-triphosphateを第二のメッセンジャ-としていることを示した。(藤井)
神経細胞(ヤリイカの巨大神経)を用いて、動的性質の異なる微小管が数種存在するが、これは構成蛋白質が異なり、また異なる機能に関与していることがわかった。(新井,小林)ウシ骨髄からneurofilamentsを精製し、再構成過程を低角度回転蒸着法を用いた電顕像により解析、neurofilamentsにおけるsubunits蛋白質の配列、重合の機序を明らかにした。(久永)
細胞膜裏打ち構造
adherence junctionを単離する方法を開発して、分子構造を明らかにした。また、protooncogeneの産物が濃縮して存在するという、興味深い成果をえた。(月田)
ウニ卵細胞分裂
細胞質分裂にあらわれる収縮環の形成過程をロ-ダミンファロイヂンによる抗体でしらべ、表層のアクチンが束ねられてつくられることを示唆する結果を得た。(馬渕)抗生物質マイトマイシンを卵内に注入すると、染色体分離が特異的に阻害されることがわかった。(浜口)

研究成果

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