Vinexin: a novel vinculin-binding protein with multiple SH3 domains enhances actin cytoskeletal organization.
Vinexin: a novel vinculin-binding protein with multiple SH3 domains enhances actin cytoskeletal organization.
复制标题
醋素：一种具有多个SH3结构域的新型Vinculin结合蛋白增强了肌动蛋白细胞骨架组织。
DOI：
10.1083/jcb.144.1.59
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发表时间：
1999-01-11
影响因子：
7.8
通讯作者：
Aota, S
中科院分区：
文献类型：
--
作者：
Kioka, N;Sakata, S;Kawauchi, T;Amachi, T;Akiyama, S K;Okazaki, K;Yaen, C;Yamada, K M;Aota, S
利用酵母双杂交系统和体外结合试验，我们已经确定了一个新的蛋白质命名为vinexin作为vinculin结合蛋白。通过北方印迹，我们确定了两种类型的vinexin mRNA的长度为3和2 kb。对全长cDNA克隆的筛选和测序表明，这两种mRNA分别编码82和37 kD的多肽，称为vinexin α和β。这两种形式的vinexin共享一个共同的羧基末端序列含有三个SH 3结构域。较大的vinexin α含有额外的氨基末端序列。vinexin和vinculin之间的相互作用是由vinexin的两个SH 3结构域和vinculin的富含脯氨酸的区域介导的。当表达时，vinexin α和β定位于NIH 3 T3成纤维细胞中的粘着斑，以及上皮LLC-PK 1细胞中的细胞-细胞连接。此外，vinexin的表达增加了粘着斑的大小。Vinexin α还促进肌动蛋白应力纤维形成的上调。此外，稳定表达vinexin β的细胞系显示增强的细胞在纤连蛋白上的铺展。这些数据将vinexin鉴定为新型粘着斑和细胞-细胞粘附蛋白，其通过SH 3结构域结合至黏着斑蛋白的铰链区，其可增强肌动蛋白细胞骨架组织和细胞铺展。
Using the yeast two-hybrid system and an in vitro binding assay, we have identified a novel protein termed vinexin as a vinculin-binding protein. By Northern blotting, we identified two types of vinexin mRNA that were 3 and 2 kb in length. Screening for full-length cDNA clones and sequencing indicated that the two mRNA encode 82- and 37-kD polypeptides termed vinexin α and β, respectively. Both forms of vinexin share a common carboxyl-terminal sequence containing three SH3 domains. The larger vinexin α contains an additional amino-terminal sequence. The interaction between vinexin and vinculin was mediated by two SH3 domains of vinexin and the proline-rich region of vinculin. When expressed, vinexin α and β localized to focal adhesions in NIH 3T3 fibroblasts, and to cell–cell junctions in epithelial LLC-PK1 cells. Furthermore, expression of vinexin increased focal adhesion size. Vinexin α also promoted upregulation of actin stress fiber formation. In addition, cell lines stably expressing vinexin β showed enhanced cell spreading on fibronectin. These data identify vinexin as a novel focal adhesion and cell– cell adhesion protein that binds via SH3 domains to the hinge region of vinculin, which can enhance actin cytoskeletal organization and cell spreading.