Role of microtubules and related proteins on cellular function

微管和相关蛋白质对细胞功能的作用

• 批准号: 60300007
• 负责人: MABUCHI Issei
• 金额: $ 10.24万
• 依托单位: University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Co-operative Research (A)
• 财政年份: 1985
• 资助国家: 日本
• 起止时间: 1985 至 1986
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-60300007/
• 关键词: Microtubule; Cytoskeleton; Actin; Chromosome Movement; Axonal Transport; Nerve Excitation; 中間径繊維

The purpose of this co-operative research project has been to clarify the mechanism of regulation of various cellular functions by cytoskeletons. There were six subprojects. 1. Function of microtubule-dynein system in mitosis The motive force for the chromosome movements was localized in the half spindle region near chromosomes. The formation of barrel-shaped spindle in the presence of T-1 was thought to be a result of scattering of the microtubule-organizing granules by T-1. MAP1 was localized on the cytoskeletal microtubules at <G_0> stage of cultured cells while it became concentrated in the nucleus at <G_1> stage. 2. Cytoskeletal proteins and cell motility Microtubules were found to undergo spontaneous assembly-disassembly cycle by dark field microscopy. Intermediate filaments were found to be relevant to mitosis and exchange of nuclei after conjugation in Tetrahymena. Complete amino acid sequences of Tetrahymena actin and of -tubulin from lymphoma cells were determined. 3. Dynamic state of microtubules in axons It was found that the 200K subunit of neurofilament is phosphorylated and transported more faster than the bulk neurofilament proteins. Axolinin was found to co-exist with microtubules in squid giant axon. 4. Function of cytoskeleton-modulating proteins A 100K-molecular weight Casensitive actin filament-severing protein was found in sea urchin eggs. Alphaactinin was found to be concentrated in the cortex of the fertilized sea urchin egg together with actin filaments. 5. Nerve excitability and microtubules Calmodulin antagonists were found to interfere with Na-current in squid nerve. 6. Cytoskeleton and cellular function Ultrastructure of the plasma membrane undercoat in skeletal muscle and astrocytes where myofibrils and intermediate filaments, respectively, attach was visualized.

该合作研究项目的目的是阐明细胞骨架调节各种细胞功能的机制。共有六个分项目。1.微管-动力蛋白系统在有丝分裂中的作用染色体运动的原动力位于靠近染色体的半纺锤体区。在T-1的存在下，桶形纺锤体的形成被认为是微管组织颗粒被T-1散射的结果。在培养细胞的阶段，MAP 1定位于细胞骨架微管上<G_0>，而在培养细胞的阶段，它变得集中在细胞核中<G_1>。2.暗视野显微镜观察发现，细胞骨架蛋白和细胞运动性微管经历自发的组装-拆卸循环。中间丝与四膜虫的有丝分裂和接合后细胞核的交换有关。完整的氨基酸序列的四膜虫肌动蛋白和微管蛋白的淋巴瘤细胞进行了测定。3.轴突中微管的动态研究发现，神经丝的200 K亚基被磷酸化，并且比大量的神经丝蛋白转运得更快。在鱿鱼巨轴突中发现Axolinin与微管共存。4.在海胆卵中发现了一种分子量为100 K的Cassensitive actin趋化蛋白。发现α辅肌动蛋白与肌动蛋白丝一起集中在受精海胆卵的皮质中。5.神经兴奋性和微管钙调素拮抗剂被发现干扰钠电流在鱿鱼神经。6.细胞骨架和细胞功能骨骼肌和星形胶质细胞中质膜底膜的超微结构，其中肌原纤维和中间丝分别附着。

项目成果

月田承一郎: Journal of Cell Biology. 102. 1710-1725 (1986)

月田城一郎：细胞生物学杂志 102。1710-1725（1986）

柿本辰男: Plant and Cell Physiology. 27. 91-101 (1986)

Tatsuo Kakimoto：植物和细胞生理学 27. 91-101 (1986)。

Kazuo Sutoh and Issei Mabuchi: "Improved method for mapping the binding site of an actin-binding protein in the actin sequence. Use of a site-directed antibody against the N-terminal region of actin as a probe of its N-terminus." Biochmistry. 25. 6186-619

Kazuo Sutoh 和 Issei Mabuchi：“改进了在肌动蛋白序列中绘制肌动蛋白结合蛋白结合位点的方法。使用针对肌动蛋白 N 末端区域的定点抗体作为其 N 末端的探针。”

Hiroshi Hosoya, Fuyuki Iwasa, Masaaki Ohnuma, Issei Mabuchi, Hideo Mohri, Hikoichi Sakai and Yukio Hiramoto: "A novel 15kDa <Ca^(2+)> -binding protein present in the eggs of the sea urchin Hemicentrotus pulcherrimus." FEBS Letters. 205. 121-126 (1986)

Hiroshi Hosoya、Fuyuki Iwasa、Masaaki Ohnuma、Issei Mabuchi、Hideo Mohri、Hikoichi Sakai 和 Yukio Hiramoto：“海胆 Hemicentrotus pulcherrimus 卵中存在一种新型 15kDa <Ca^(2 )> 结合蛋白。”

Takako Nishi, Makoto Kobayashi and Yoshinobu Shigenaka: "Membrane activity and its correlation with vacuolar contraction in the Heliozoan Echinosphaerium." Journal of Experimental Zoology. 239. 175-182 (1986)

Takako Nishi、Makoto Kobayashi 和 Yoshinobu Shigenaka：“日光虫棘球藻中的膜活性及其与液泡收缩的相关性。”