title: Nck enables directional cell migration through the coordination of polarized membrane protrusion with adhesion dynamics.
abstract: 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 α5β1-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.
authors: Department of Veterinary Pathobiology, Texas A&M University, College Station, TX 77843-4467, USA.
authors: Sankar P
authors: SP
authors: Chaki
authors: Rola
authors: R
authors: Barhoumi
authors: Matthew E
authors: ME
authors: Berginski
authors: Harini
authors: H
authors: Sreenivasappa
authors: Andreea
authors: A
authors: Trache
authors: Shawn M
authors: SM
authors: Gomez
authors: Gonzalo M
authors: GM
authors: Rivera
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism*; D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D002448:Cell Adhesion / Q000235:genetics / Q000502:physiology; D002460:Cell Line; D002465:Cell Movement / Q000235:genetics / Q000502:physiology*; D016764:Cell Polarity / Q000235:genetics / Q000502:physiology*; D022001:Focal Adhesions / Q000378:metabolism; D006801:Humans; D039081:Integrin alpha5beta1 / Q000378:metabolism; D051379:Mice; D018625:Microscopy, Atomic Force; D008856:Microscopy, Fluorescence; D041681:NIH 3T3 Cells; D015513:Oncogene Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D039081:Integrin alpha5beta1; C078332:Nck protein; D015513:Oncogene Proteins
keywords: Actin cytoskeleton; Cell adhesion; Cell polarity; Directional migration; Nck