Molecular mechanism of cytokinesis in animal cells

动物细胞胞质分裂的分子机制

• 批准号: 06404004
• 负责人: MABUCHI Issei
• 金额: $ 19.97万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06404004/
• 关键词: cytokinesis; contractile ring; actin filament; cleavage signal; rho; actin-modulating protein; egg; fission yeast; アクチン; ミオシン; リン酸化; 分裂溝; 低分子量Gタンパク質

1. Cleavage signaling pathwayWe found a new rho gene from the fission yeast and named it rho5^+.Disruptant of rho1 gene could not grow indicating that rho1^+ is an essential gene. We localized Rho1 by both immunofluorescence microscopy and expression of GFP-Rhol fusion protein in the fission yeast cell to growing tips during interphase and to septum during cytokinesis. Overexperssion of Rhol induced aberrant morphology, delocalization of actin patches, and thickening of cell wall. Overexperssion of Rho2 was lethal, cell morphology was aberrant and cell wall was abnormally thick. Disruption of rho3 gene induced aberrant morphology of the cell and abnormal division. Disruption of rho4 gene induced loss of cell polarity and abnormal division.2. Structure and foumation of the contractile ringWe found novel F-actin-binding proteins from sea urchin eggs by F-actin affinity chromatography. Among them, 60K protein was indentified as coronin and 150K protein was identified as myosin 6. Immunofluorescence microscopy showed that these proteins and ARP3, which we indentified in the last year, are present in the cleavage furrow cortex. We gene-cloned ARP3 from fission yeast and disrupted it. The terminal phenotype showed that actin patches were delocalized. ABP40, a protein which is concentrated in the cleavage furrow of the sea urchin egg, was demonstrated to have an F-actin-severing activity.

1. 我们从分裂酵母中发现了一个新的rho基因，命名为rho5^+。rho1基因的干扰物不能生长，表明rho1^+是必需基因。我们通过免疫荧光显微镜和GFP-Rhol融合蛋白在分裂酵母细胞中的表达，将Rho1定位在间期生长尖端和细胞质分裂期间的隔膜上。Rhol基因的过度表达导致细胞形态异常、肌动蛋白斑块脱位和细胞壁增厚。Rho2过表达致死性，细胞形态异常，细胞壁异常厚。rho3基因的破坏导致细胞形态异常和分裂异常。rho4基因的破坏导致细胞极性丧失和异常分裂。通过F-actin亲和层析，从海胆卵中发现了新的F-actin结合蛋白。其中60K蛋白鉴定为冠蛋白，150K蛋白鉴定为肌凝蛋白6。免疫荧光显微镜显示，这些蛋白质和ARP3，我们在去年鉴定，存在于卵裂沟皮层。我们从裂变酵母中克隆了ARP3基因并破坏了它。末端表型显示肌动蛋白斑块脱位。ABP40是一种集中在海胆卵卵裂沟中的蛋白质，被证明具有切断f -actin的活性。

项目成果

Yokota,E.,Mabuchi,I.: "Interaction of flagellar inner arem dynein isolated from sea urchin sperm with microtubules in the presence of ATP." Europearn J.Cell Biology. in press.(1997)

Yokota,E.,Mabuchi,I.：“在 ATP 存在下，从海胆精子中分离出的鞭毛内部动力蛋白与微管的相互作用。”

Kojima, S., Mishima, M., Mabuchi, I.and Hotta, Y.: "A Single Drosophila melanogaster myosin light chain kinase gene produces multiple isoforms whose activities are differently regulated." Genes to Cells. 1. 855-871 (1996)

Kojima, S.、Mishima, M.、Mabuchi, I. 和 Hotta, Y.：“单个果蝇肌球蛋白轻链激酶基因产生多种同工型，其活性受到不同的调节。”

Issei Mabuchi: "Cleavage furrow:timing of emergence of contractilering actin filaments and establish ment of the contractilering." Journal of Cell Science. 107. 1853-1862 (1994)

Issei Mabuchi：“分裂沟：收缩肌动蛋白丝的出现和收缩的建立的时间。”

Kojima,S.,Mishima,M.: "A single Drosophila melanogaster myosin light chain kinase gene produces multiple isoforms whose activities are..." Genes to Cells. 1. 855-871 (1996)

Kojima,S.,Mishima,M.：“单个果蝇肌球蛋白轻链激酶基因产生多种亚型，其活性是......”基因到细胞。

Yokota, E.and Mabuchi, I.: "Interaction of flagellar inner arm dynein isolated from sea urchin sperm with microtubules in the presence of ATP." Europearn J.Cell Bilogy. (in press). (1977)

Yokota, E. 和 Mabuchi, I.：“在 ATP 存在下，从海胆精子中分离出的鞭毛内臂动力蛋白与微管的相互作用。”