Analysis of molecular mechanisms of cell division in Tetrahymena using cell-division-arrest mutants.

使用细胞分裂停滞突变体分析四膜虫细胞分裂的分子机制。

• 批准号: 02454020
• 负责人: WATANABE Yoshio
• 金额: $ 4.29万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-02454020/
• 关键词: Tetrahymena; cell division; cytokinesis; cell-division mutant; actin; profilin; EF-1alpha; calcium-binding protein; 分裂停止突然変異株; テトラヒメナ・アクチン; Ca^<2+>-結合蛋白質; 分裂停止突然変異; 収縮環; DNA複製; EFー1α; テトラヒメナ アクチン; テトラヒメナ プロフィリン; カルシウム 結合蛋白質; アンチセンス RNA /

To elucidate the molecular mechanisms of cell division in animal cells,by using Tetrahymena's cell-division-arrest mutants, cdaA and cdaC, we have analyzed mechanisms for division plane determination, division furrow formation, furrowing, contraction-related contractle ring microfilament disassembly, and for calcium-regulation of division furrow constriction.Concerning division plane determination, it was evident that cdaA mutant gene product,p85, plaied a crucial role in the determination and it served as polymerization nuclei of the contractile ring actin filaments. As for division furrow constriction, by the analysis of cdaC mutant, the mutant gene product is considered to be an actin-binding protein capable of bundling the contractile ring microfilaments. We recently found a protein which associates with Tetrahymena's 14-nm filaments and we identified it as EF-1alpha from its gene analysis. We are now investigating the relation between EF-1alpha and cdaC gene product, since Tetrahymena EF-1alpha has an actin-bundling activity. In cytokinesis, it has been proposed that contractile ring microfilament disassembly occurs accompanying with the contraction of contractile ring. In this regard, we found that an actin-polymerization-inhibitory factor, profilin of Tetrahymena, colocalized with actin filaments in the division furrow, suggesting the involvement of profilin in the formation and disassembly of contractile ring microfilaments. Concerning the calcium-regulation of division furrow constriction, we cloned the genes for 3 kinds of calcium-binding proteins (calmodulin, TCBP-23, and TCBP-25) from Tetrahymena. We are now scrutinized their biological functions in cytokinesis by injecting the gene or modified gene products yielded in E. coli.

为了阐明动物细胞中细胞分裂的分子机制，使用四膜虫的细胞 - 分段 - 候选突变体，CDAA和CDAC，我们分析了分裂平面测定，分裂形成，皱纹，与收缩相关的合同环形拆卸和相关的合同环的拆卸，以及分裂平面的机制为了使分裂平面的确定钙调节，很明显，CDAA突变基因产物P85在确定中起着至关重要的作用，并且它是收缩环肌动蛋白丝的聚合核。至于分裂犁沟收缩，通过对CDAC突变体的分析，突变基因产物被认为是能够捆绑收缩环微丝的肌动蛋白结合蛋白。我们最近发现了一种与四膜虫的14 nm丝相关的蛋白质，并通过其基因分析将其确定为EF-1Alpha。我们现在正在研究EF-1Alpha和CDAC基因产物之间的关系，因为四环EF-1Alpha具有肌动蛋白捆绑活性。在细胞因子中，有人提出收缩环微丝拆卸发生，伴随着收缩环的收缩。在这方面，我们发现肌动蛋白 - 聚合抑制因子，四环烷的profilin与肌动蛋白丝中的分裂沟中共定位，这表明叶酸蛋白参与了收缩环微丝的形成和分解。关于分裂犁沟收缩的钙调节，我们将三种钙结合蛋白（钙调蛋白，TCBP-23和TCBP-25）的基因克隆起来。现在，我们通过注射大肠杆菌中产生的基因或改性基因产物来仔细检查其在细胞因子中的生物学功能。

项目成果

Numata,O.,Takemasa,T.,Takagi,I.,Hirono,M.,Hirano,H.,Chiba,J. & Watanabe,Y.: "Tetrahymena 14-nm filament-forming protein has citrate synthase activity" Biochem. Biophys. Res. Commun.174. 1028-1034 (1991)

沼田，O.，武正，T.，高木，I.，广野，M.，平野，H.，千叶，J.

Takemasa,T.,Takagi,N.and Watanabe,Y.: "Calciumーbinding proteins from Tetrahymena.In Novel CalciumーBinding Proteins.Fundamentals and Clinical Implications.,ed.by C.W.Heizman,Chapter26" Springer Press, 481-495 (1991)

Takemasa, T.、Takagi, N. 和 Watanabe, Y.：“来自四膜虫的钙结合蛋白。新型钙结合蛋白。基本原理和临床意义。”，C.W. Heizman 编辑，第 26 章”Springer Press，481- 495 (1991)

Numata,O.,Tomiyoshi,T.,Kurasawa,Y.,Fu,Z-x., Takahashi,M.,Kosaka,T.,Chiba,J. & Watanabe,Y.: "Antibodies against Tetrahymena 14-nm filament-forming protein recognize the replication band in Euplotes" Exptl. Cell Res.193. 183-189 (1991)

沼田，O.，富吉，T.，仓泽，Y.，Fu，Z-x.，高桥，M.，小坂，T.，千叶，J.

Hirono,M.,Tanaka,R.,& Watanabe,Y.: "<Tetrahymena>___ー actin:Copolymerization with skeletal muscle actin and interactions with muscle actinーbinding proteins." J.Biochem.107. 32-36 (1990)

Hirono, M.、Tanaka, R. 和 Watanabe, Y.：“<四膜虫>___- 肌动蛋白：与骨骼肌肌动蛋白的共聚以及与肌肉肌动蛋白结合蛋白的相互作用。J.Biochem.107（1990 年）。 ）

Watanabe,Y.,Hirono,M.,Takemasa,T.,& Numata,O: "Protozoan genes:As viewed from five <Tetrahymena>___ー genes." Japan.J.Protozool.23. 1-11 (1990)

Watanabe, Y., Hirono, M., Takemasa, T., & Numata, O：“原生动物基因：从五个 <四膜虫>___- 基因来看。Japan.J.Protozool.23 (1990)。