期刊
EXPERIMENTAL CELL RESEARCH
卷 316, 期 11, 页码 1829-1844出版社
ELSEVIER INC
DOI: 10.1016/j.yexcr.2010.02.007
关键词
Actin; Intermediate filaments; Cell adhesion; Cell guidance
资金
- Canadian Institutes of Health Research [MGP-37783]
- Group and Research Resource
- Heart and Stroke Foundation of Canada
- MiCInn, Spain [SAF2006-03489]
Cell adhesion and spreading on collagen, which are essential processes for development and wound healing in mammals, are mediated by beta 1 integrins and the actin and intermediate filament cytoskeletons. The mechanisms by which these separate cytoskeletal systems interact to regulate beta 1 integrins and cell spreading are poorly defined. We previously reported that the actin cross-linking protein filamin A binds the intermediate filament protein vimentin and that these two proteins co-regulate cell spreading. Here we used deletional mutants of filamin A to define filamin A-vimentin interactions and the subsequent phosphorylation and re-distribution of vimentin during cell spreading on collagen. Imaging of fixed and live cell preparations showed that phosphorylated vimentin is translocated to the cell membrane during spreading. Knockdown of filamin A inhibited cell spreading and the phosphorylation and re-distribution of vimentin. Knockdown of filamin A and/or vimentin reduced the cell surface expression and activation of beta 1 integrins, as indicated by immunoblotting of plasma membrane-associated proteins and shear force assays. In vitro pull-down assays using filamin A mutants showed that both vimentin and protein kinase C epsilon bind to repeats 1-8 of filamin A. Reconstitution of filamin-A-deficient cells with full-length filamin A or filamin A repeats 1-8 restored cell spreading, vimentin phosphorylation, and the cell surface expression of beta 1 integrins. We conclude that the binding of filamin A to vimentin and protein kinase C epsilon is an essential regulatory step for the trafficking and activation of beta 1 integrins and cell spreading on collagen. (C) 2010 Elsevier Inc. All rights reserved.
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