Mingle Lisa A, Okuhama Nataly N, Shi Jian, Singer Robert H, Condeelis John, Liu Gang
Center for Cell Biology and Cancer Research, Albany Medical College, 47 New Scotland Avenue, Albany, NY 12208, USA.
J Cell Sci. 2005 Jun 1;118(Pt 11):2425-33. doi: 10.1242/jcs.02371.
The actin-related protein 2/3 (Arp2/3) complex is a crucial actin polymerization nucleator and is localized to the leading protrusions of migrating cells. However, how the multiprotein complex is targeted to the protrusions remains unknown. Here, we demonstrate that mRNAs for the seven subunits of the Arp2/3 complex are localized to the protrusions in fibroblasts, supporting a hypothesis that the Arp2/3 complex is targeted to its site of function by mRNA localization. Depletion of serum from culture medium inhibits Arp2/3-complex mRNA localization to the protrusion, whereas serum stimulation leads to significant mRNA localization within 30 minutes. The effect of serum suggests that Arp2/3-complex mRNA localization is a cellular response to extracellular stimuli. The localization of the Arp2/3 complex mRNAs is dependent on both actin filaments and microtubules, because disruption of either cytoskeletal system (with cytochalasin D and colchicine, respectively) inhibited the localization of all seven subunit mRNAs. In addition, myosin inhibitors significantly inhibit Arp2 mRNA localization in chicken embryo fibroblasts, suggesting a myosin motor dependent mechanism for Arp2/3-complex mRNA localization.
肌动蛋白相关蛋白2/3(Arp2/3)复合体是一种关键的肌动蛋白聚合成核因子,定位于迁移细胞的前沿突起处。然而,这种多蛋白复合体如何靶向到突起部位仍不清楚。在这里,我们证明Arp2/3复合体七个亚基的mRNA定位于成纤维细胞的突起处,支持了一种假说,即Arp2/3复合体通过mRNA定位靶向到其功能位点。从培养基中去除血清会抑制Arp2/3复合体mRNA向突起部位的定位,而血清刺激则会在30分钟内导致显著的mRNA定位。血清的作用表明Arp2/3复合体mRNA定位是细胞对细胞外刺激的一种反应。Arp2/3复合体mRNA的定位依赖于肌动蛋白丝和微管,因为破坏任何一种细胞骨架系统(分别用细胞松弛素D和秋水仙碱)都会抑制所有七个亚基mRNA的定位。此外,肌球蛋白抑制剂显著抑制鸡胚成纤维细胞中Arp2 mRNA的定位,提示存在一种依赖肌球蛋白马达的Arp2/3复合体mRNA定位机制。