Probing the integrin-actin linkage using high-resolution protein velocity mapping

• DOI: 10.1242/jcs.03321
• Publication Date: 2006-12-11T22:44:32Z
• Authors: Claire M. Brown; Benedict Hebert; David L. Kolin; Jessica Zareno; Leanna Whitmore; Alan Rick Horwitz; Paul W. Wiseman
• Keywords: 0301 basic medicine; Cytoplasm; Integrins; Green Fluorescent Proteins; 3T3 Cells; CHO Cells; Models, Biological; Actins; Cytoskeletal Proteins; Luminescent Proteins; Mice; 03 medical and health sciences; Cricetulus; Cricetinae; Image Interpretation, Computer-Assisted; Protein Interaction Mapping; Cell Adhesion; Image Processing, Computer-Assisted; Animals; Algorithms; Cells, Cultured; Cytoskeleton

Cell migration is regulated in part by the connection between the substratum and the actin cytoskeleton. However, the very large number of proteins involved in this linkage and their complex network of interactions make it difficult to assess their role in cell migration. We apply a novel image analysis tool, spatio-temporal image correlation spectroscopy (STICS), to quantify the directed movements of adhesion-related proteins and actin in protrusions of migrating cells. The STICS technique reveals protein dynamics even when protein densities are very low or very high, and works in the presence of large, static molecular complexes. Detailed protein velocity maps for actin and the adhesion-related proteins α-actinin, α5-integrin, talin, paxillin, vinculin and focal adhesion kinase are presented. The data show that there are differences in the efficiency of the linkage between integrin and actin among different cell types and on the same cell type grown on different substrate densities. We identify potential mechanisms that regulate efficiency of the linkage, or clutch, and identify two likely points of disconnect, one at the integrin and the other at α-actinin or actin. The data suggests that the efficiency of the linkage increases as actin and adhesions become more organized showing the importance of factors that regulate the efficiency in adhesion signaling and dynamics.