Involvement of the Arp2/3 complex and Scar2 in Golgi polarity in scratch wound models
Involvement of the Arp2/3 complex and Scar2 in Golgi polarity in scratch wound models
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DOI：
10.1091/mbc.e02-06-0345
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发表时间：
2003-02-01
影响因子：
3.3
通讯作者：
Machesky, LM
中科院分区：
文献类型：
--
作者：
Magdalena, J;Millard, TH;Machesky, LM
细胞运动性和细胞极性对于形态发生、免疫系统功能和组织修复是必不可少的。许多动物细胞通过爬行运动，运动的一个主要驱动力来自肌动蛋白丝的协调组装和拆卸。当组织培养细胞迁移以闭合划痕伤口时，这种定向延伸伴随着高尔基体重新定向，以面向伤口前缘，从而使运动细胞的固有极性相对于伤口边缘和细胞迁移方向对齐。Rho家族GTP酶下游的肌动蛋白聚合所必需的细胞蛋白质包括作为肌动蛋白成核剂的Arp 2/3复合物和作为Arp 2/3复合物的激活剂的Wiskott-Aldrich综合征蛋白（WASP）家族的成员。因此，我们分析了参与的Arp 2/3复合物和WASP家族蛋白在体外伤口愈合试验中使用NIH 3 T3成纤维细胞和星形胶质细胞。在NIH 3 T3细胞中，我们发现肌动蛋白和Arp 2/3复合物有助于细胞极性的建立。此外，N-末端片段的Scar 2（而不是N-WASP或Scar 1或Scar 3）的过表达干扰NIH 3 T3高尔基体极化，但不与细胞迁移。与此相反，肌动蛋白，Arp 2/3，和WASP家族蛋白似乎没有参与星形胶质细胞的高尔基体极化。因此，我们的研究结果表明，高尔基体极性建立的要求是细胞类型特异性的。此外，在NIH 3 T3细胞中，Scar 2和Arp 2/3复合物似乎参与定向迁移过程中高尔基体极性的建立和维持。
Cell motility and cell polarity are essential for morphogenesis, immune system function, and tissue repair. Many animal cells move by crawling, and one main driving force for movement is derived from the coordinated assembly and disassembly of actin filaments. As tissue culture cells migrate to close a scratch wound, this directional extension is accompanied by Golgi apparatus reorientation, to face the leading wound edge, giving the motile cell inherent polarity aligned relative to the wound edge and to the direction of cell migration. Cellular proteins essential for actin polymerization downstream of Rho family GTPases include the Arp2/3 complex as an actin nucleator and members of the Wiskott-Aldrich Syndrome protein (WASP) family as activators of the Arp2/3 complex. We therefore analyzed the involvement of the Arp2/3 complex and WASP-family proteins in in vitro wound healing assays using NIH 3T3 fibroblasts and astrocytes. In NIH 3T3 cells, we found that actin and Arp2/3 complex contributed to cell polarity establishment. Moreover, overexpression of N-terminal fragments of Scar2 (but not N-WASP or Scar1 or Scar3) interfere with NIH 3T3 Golgi polarization but not with cell migration. In contrast, actin, Arp2/3, and WASP-family proteins did not appear to be involved in Golgi polarization in astrocytes. Our results thus indicate that the requirement for Golgi polarity establishment is cell-type specific. Furthermore, in NIH 3T3 cells, Scar2 and the Arp2/3 complex appear to be involved in the establishment and maintenance of Golgi polarity during directed migration.