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Anchoring mechanism of the intracellulan organization of actin cytoskeleton by the cell wall.

细胞壁对肌动蛋白细胞骨架的细胞内组织的锚定机制。

基本信息

批准号：
08640827
负责人：
TAKAGI Shingo
金额：
$ 1.28万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08640827/
关键词：
actin cytoskeleton cell wall cytoplasmic streaming mesophyll cell microfilament RGD motif RYD motif Vallisneria

项目摘要

In mesophyll cells of an aquatic angiosperm Vallisneria gigantea, bundles of actin filaments serve as the tracks for rotational streaming of the cytoplasm. From the previous studies, we hypothesized that the cell wall plays a crucial role to maintain the intracellular organization of the bundles of actin filaments through the plasma membrane as seen in the structure of focal adhesion in animal cells. Exogenously applied RGD or RYD hexapeptide caused a drastic disorder in the pattern of cytoplasmic streaming as well as in the intracellular array of actin filaments. Polyclonal antibodies raised against the RGD motif detected two polypeptides, 54 and 27 kDa, in the whole extract from the leaves, which were localized in the cell wall by immunofluorescence microscopy. Putative protein factors for these polypeptides were searched in highly purified plasma membrane fraction by affinity chromatography using the RGD peptide as a ligand. The eluted fraction with EDTA solution contained protein factors, whereas the amount of those was too small to be resolved after SDS PAGE.To know intracellular factors involved, the mode of disturbance of the tracks of cytoplasmic streaming was observed in a living mesophyll cell by video microscopy during plasmolysis and deplasmolysis. The bundle structure of the tracks seemed to be ramified only transiently. Even after recovery of the structure of bundles, the pattern of cytoplasmic streaming was unfixed and continued to change dynamically. These observations suggested the involvement of intracellular factors that bundle the actin filaments and fix the bundles to the plasma membrane and/or the cell wall.

在水生被子植物苦草的叶肉细胞中，成束的肌动蛋白丝作为细胞质旋转流动的轨道。从以前的研究中，我们假设细胞壁起着至关重要的作用，以维持细胞内的肌动蛋白丝束通过质膜的组织中所看到的在动物细胞中的粘着斑的结构。外源性应用的RGD或赖德六肽引起的细胞质流的模式，以及在细胞内阵列的肌动蛋白丝的剧烈紊乱。针对RGD基序的多克隆抗体检测到两个多肽，54和27 kDa，在整个提取物的叶子，这是本地化的细胞壁通过免疫荧光显微镜。这些多肽的推定的蛋白质因子进行了搜索，在高度纯化的质膜馏分通过亲和层析使用的RGD肽作为配体。用EDTA溶液洗脱的组分中含有蛋白质因子，而SDS PAGE后这些蛋白质因子的量太少而不能分辨.为了了解细胞内因子的参与，我们用视频显微镜观察了活叶肉细胞在质体分裂和去质体分裂过程中胞质流动轨迹的扰动模式.轨道的束结构似乎只是暂时的分支。即使在恢复束的结构，胞质流的模式是不固定的，并继续动态变化。这些观察结果表明，参与细胞内的因素，捆绑肌动蛋白丝和固定束质膜和/或细胞壁。