CNRS UPR3082, Laboratoire d'Enzymologie et Biochimie Structurales, Avenue de la Terrasse, 91198 Gif-sur-Yvette Cedex, France.
J Cell Sci. 2011 Oct 15;124(Pt 20):3414-27. doi: 10.1242/jcs.081083.
The Scar/Wave complex (SWC) generates lamellipodia through Arp2/3-dependent polymerisation of branched actin networks. In order to identify new SWC regulators, we conducted a screen in Drosophila cells combining proteomics with functional genomics. This screen identified Clathrin heavy chain (CHC) as a protein that binds to the SWC and whose depletion affects lamellipodium formation. This role of CHC in lamellipodium formation can be uncoupled from its role in membrane trafficking by several experimental approaches. Furthermore, CHC is detected in lamellipodia in the absence of the adaptor and accessory proteins of endocytosis. We found that CHC overexpression decreased membrane recruitment of the SWC, resulting in reduced velocity of protrusions and reduced cell migration. By contrast, when CHC was targeted to the membrane by fusion to a myristoylation sequence, we observed an increase in membrane recruitment of the SWC, protrusion velocity and cell migration. Together these data suggest that, in addition to its classical role in membrane trafficking, CHC brings the SWC to the plasma membrane, thereby controlling lamellipodium formation.
瘢痕/波复合物(SWC)通过 Arp2/3 依赖性分支肌动蛋白网络聚合产生片状伪足。为了鉴定新的 SWC 调节剂,我们在果蝇细胞中进行了一项结合蛋白质组学和功能基因组学的筛选。该筛选鉴定出网格蛋白重链(CHC)作为一种与 SWC 结合的蛋白质,其耗竭会影响片状伪足的形成。通过几种实验方法,可以将 CHC 在片状伪足形成中的作用与它在膜运输中的作用分离。此外,在没有内吞作用的衔接蛋白和辅助蛋白的情况下,CHC 也会被检测到在片状伪足中。我们发现 CHC 的过表达会减少 SWC 向膜的募集,导致突起速度降低和细胞迁移减少。相比之下,当 CHC 通过与豆蔻酰化序列融合被靶向到膜上时,我们观察到 SWC 向膜的募集、突起速度和细胞迁移增加。这些数据表明,CHC 除了在膜运输中的经典作用外,还将 SWC 带到质膜,从而控制片状伪足的形成。