Nck enables directional cell migration through the coordination of polarized membrane protrusion with adhesion dynamics

被引:41
作者
Chaki, Sankar P. [1 ]
Barhoumi, Rola [2 ]
Berginski, Matthew E. [3 ]
Sreenivasappa, Harini [4 ]
Trache, Andreea [4 ]
Gomez, Shawn M. [3 ]
Rivera, Gonzalo M. [1 ]
机构
[1] Texas A&M Univ, Dept Vet Pathobiol, College Stn, TX 77843 USA
[2] Texas A&M Univ, Dept Vet Integrat Biosci, College Stn, TX 77843 USA
[3] Univ N Carolina, Dept Biomed Engn, Chapel Hill, NC 27599 USA
[4] Texas A&M Univ, Hlth Sci Ctr, Dept Syst Biol & Translat Med, College Stn, TX 77843 USA
基金
美国国家科学基金会;
关键词
Nck; Cell adhesion; Actin cytoskeleton; Cell polarity; Directional migration; RECEPTOR TYROSINE KINASES; ACTIN-BASED MOTILITY; N-WASP; ADAPTER PROTEINS; MYOSIN-II; RUFFLE FORMATION; CRUCIAL ROLE; ACTIVATION; COMPLEX; BETA;
D O I
10.1242/jcs.119610
中图分类号
Q2 [细胞生物学];
学科分类号
071013 [干细胞生物学];
摘要
Directional migration requires the coordination of cytoskeletal changes essential for cell polarization and adhesion turnover. Extracellular signals that alter tyrosine phosphorylation drive directional migration by inducing reorganization of the actin cytoskeleton. It is recognized that Nck is an important link between tyrosine phosphorylation and actin dynamics; however, the role of Nck in cytoskeletal remodeling during directional migration and the underlying molecular mechanisms remain largely undetermined. In this study, a combination of molecular genetics and quantitative live cell microscopy was used to show that Nck is essential in the establishment of front-back polarity and directional migration of endothelial cells. Time-lapse differential interference contrast and total internal reflection fluorescence microscopy showed that Nck couples the formation of polarized membrane protrusions with their stabilization through the assembly and maturation of cell-substratum adhesions. Measurements by atomic force microscopy showed that Nck also modulates integrin alpha 5 beta 1-fibronectin adhesion force and cell stiffness. Fluorescence resonance energy transfer imaging revealed that Nck depletion results in delocalized and increased activity of Cdc42 and Rac. By contrast, the activity of RhoA and myosin II phosphorylation were reduced by Nck knockdown. Thus, this study identifies Nck as a key coordinator of cytoskeletal changes that enable cell polarization and directional migration, which are crucial processes in development and disease.
引用
收藏
页码:1637 / 1649
页数:13
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