Isolation of a novel 190 kDa protein from tobacco BY-2 cells: Possible involvement in the interaction between actin filaments and microtubules
Isolation of a novel 190 kDa protein from tobacco BY-2 cells: Possible involvement in the interaction between actin filaments and microtubules
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DOI：
10.1093/pcp/pcd015
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发表时间：
2000-08-01
影响因子：
4.9
通讯作者：
Sonobe, S
中科院分区：
文献类型：
--
作者：
Igarashi, H;Orii, H;Sonobe, S
肌动蛋白丝（actin filaments，AFs）和微管（microtubules，MTs）之间的相互作用已在多种植物细胞中被报道，并且已被认为存在连接这两种细胞骨架网络的因子，但其分子实体尚未阐明。我们得到了一个馏分含有MT结合多肽，诱导捆绑的AF和MT。一个190 kDa的多肽，与AF选择性地从该馏分中分离。这种多肽被认为具有结合AF和MT两者的能力。我们提出了一种针对190 kDa多肽的单克隆抗体。免疫染色表明，190 kDa的多肽与AF束和MT束在体外形成的协会。整个细胞周期的免疫细胞化学研究表明，190 kDa的多肽被定位在核被膜破裂前的细胞核中，并在纺锤体和成膜细胞分裂过程中。在核膜重新形成后，190 kDa多肽被隔离到子核。使用该抗体，我们成功地克隆了编码190 kDa多肽的cDNA。
Interaction between actin filaments (AFs) and microtubules (MTs) has been reported in various plant cells, and the presence of a factor(s) connecting these two cytoskeletal networks has been suggested, but its molecular entity has not been elucidated yet. We obtained a fraction containing MT-binding polypeptides, which induced bundling of AFs and of MTs. A 190 kDa polypeptide which associated with AFs was selectively isolated from the fraction. This polypeptide was thought to have an ability to bind to both AFs and MTs. We raised a monoclonal antibody against the 190 kDa polypeptide. Immunostaining demonstrated the association of the 190 kDa polypeptide with AF bundles and with MT bundles formed in vitro. Immunocytochemical studies throughout the cell cycle revealed that the 190 kDa polypeptide was localized in the nucleus before nuclear envelope breakdown, and in the spindle and the phragmoplast during cell division. After the re-formation of the nuclear envelope, the 190 kDa polypeptide was sequestered to the daughter nuclei. Using the antibody we succeeded in cloning a cDNA encoding the 190 kDa polypeptide.