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MBC in Press, published online ahead of print July 19, 2006
Mol. Biol. Cell 10.1091/mbc.E06-06-0496

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Submitted on June 6, 2006
Revised on June 26, 2006
Accepted on July 11, 2006

Initiation of Attachment and Generation of Mature Focal Adhesions by Integrin-containing Filopodia in Cell Spreading

Michael A. Partridge and Eugene E. Marcantonio

Department of Pathology and Cell Biology, College of Physicians and Surgeons, Columbia University, New York, NY 10032

Monitoring Editor: Richard Assoian

Integrin receptors, and associated cytoplasmic proteins mediate adhesion, cell signaling and connections to the cytoskeleton. Using fluorescent protein chimeras, we analyzed initial integrin adhesion in spreading fibroblasts with Total Internal Reflection Fluorescence (TIRF) microscopy. Surprisingly, sequential radial projection of integrin and actin containing filopodia formed the initial cell-matrix contacts. These Cdc42-dependent, integrin-containing projections recruited cytoplasmic focal adhesion (FA) proteins in a hierarchical manner; initially talin with integrin and subsequently FAK and paxillin. Radial FA structures then anchored cortical actin bridges between them and subsequently cells reorganized their actin, a process promoted by Src, and characterized by lateral FA reorientation to provide anchor points for actin stress fibers. Finally, the nascent adhesions coalesced until they formed mature FAs.

Address correspondence to: Eugene E. Marcantonio (eem2{at}columbia.edu)




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