title: Interaction of crk with Myosin-1c participates in fibronectin-induced cell spreading.
abstract: We previously reported a novel interaction between v-Crk and myosin-1c, and demonstrated that this interaction is essential for cell migration, even in the absence of p130CAS. We here demonstrate a role for Crk-myosin-1c interaction in cell adhesion and spreading. Crk-knockout (Crk ⁻/⁻) mouse embryo fibroblasts (MEFs) exhibited significantly decreased cell spreading and reduced Rac1 activity. A stroboscopic analysis of cell dynamics during cell spreading revealed that the cell-spreading deficiency in Crk⁻/⁻ MEFs was due to the short protrusion/retraction distances and long persistence times of membrane extensions. The low activity of Rac1 in Crk⁻/⁻ MEFs, which led to delayed cell spreading in these cells, is consistent with the observed defects in membrane dynamics. Reintroduction of v-Crk into Crk⁻/⁻ MEFs rescued these defects, restoring cell-spreading activity and membrane dynamics to Crk⁺/⁺ MEF levels, and normalizing Rac1 activity. Knockdown of myosin-1c by introduction of small interfering RNA resulted in a delay in cell spreading and reduced Rac1 activity to low levels, suggesting that myosin-1c also plays an essential role in cell adhesion and spreading. In addition, deletion of the v-Crk SH3 domain, which interacts with the myosin-1c tail, led to defects in cell spreading. Overexpression of the GFP-myosin-1c tail domain effectively inhibited the v-Crk-myosin-1c interaction and led to a slight decrease in cell spreading and cell surface area. Collectively, these findings suggest that the v-Crk-myosin-1c interaction, which modulates membrane dynamics by regulating Rac1 activity, is crucial for cell adhesion and spreading.
authors: Bio Imaging and Cell Dynamics Research Center, School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 500-712, Korea.
authors: Hyejin
authors: H
authors: Oh
authors: Hwan
authors: H
authors: Kim
authors: Baehyun
authors: B
authors: Shin
authors: Kun Ho
authors: KH
authors: Lee
authors: Myeong Gu
authors: MG
authors: Yeo
authors: Woo Keun
authors: WK
authors: Song
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000235:genetics / Q000502:physiology*; D002462:Cell Membrane / Q000502:physiology*; D002465:Cell Movement / Q000235:genetics / Q000502:physiology*; D017931:DNA Primers / Q000235:genetics; D005347:Fibroblasts; D005353:Fibronectins / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D015151:Immunoblotting; D007150:Immunohistochemistry; D051379:Mice; D018345:Mice, Knockout; D024461:Myosin Type I / Q000235:genetics / Q000378:metabolism*; D009479:Neuropeptides / Q000378:metabolism; D050737:Proto-Oncogene Proteins c-crk / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D049489:Stroboscopy; D020830:rac1 GTP-Binding Protein / Q000378:metabolism
chemical_list: C476798:Crk protein, mouse; D017931:DNA Primers; D005353:Fibronectins; C470422:Myo1c protein, mouse; D009479:Neuropeptides; D050737:Proto-Oncogene Proteins c-crk; D034741:RNA, Small Interfering; C484602:Rac1 protein, mouse; D024461:Myosin Type I; D020830:rac1 GTP-Binding Protein
keywords: cell adhesion; cell spreading; myosin-1c; v-Crk
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