Human p21-activated kinase (Pak1) regulates actin organization in mammalian cells

• Published in: Current biology 1997-03, Vol.7 (3), p.202-210
• Main Authors: Sells, Mary Ann, Knaus, Ulla G, Bagrodia, Shubha, Ambrose, Diane M, Bokoch, Gary M, Chernoff, Jonathan
• Format: Article
• Language: English
• Subjects: 3T3 Cells - drug effects; 3T3 Cells - ultrastructure; Actin Cytoskeleton - metabolism; Actin Cytoskeleton - ultrastructure; Actins - metabolism; Actins - ultrastructure; Adaptor Proteins, Signal Transducing; Animals; cdc42 GTP-Binding Protein, Saccharomyces cerevisiae; Cell Cycle Proteins - metabolism; Cell Membrane - ultrastructure; Cell Movement; COS Cells; Cytoskeleton - metabolism; Cytoskeleton - ultrastructure; Glutathione Transferase - metabolism; GTP-Binding Proteins - metabolism; Humans; Mice; Microinjections; Models, Biological; Oncogene Proteins - metabolism; p21-Activated Kinases; Protein Binding; Protein-Serine-Threonine Kinases - pharmacology; Protein-Serine-Threonine Kinases - physiology; rac GTP-Binding Proteins; Recombinant Fusion Proteins - metabolism; src Homology Domains; Vinculin - metabolism
• ISSN: 0960-9822; 1879-0445
• DOI: 10.1016/S0960-9822(97)70091-5

Background: The Rho family GTPases Cdc42, Rac1 and RhoA regulate the reorganization of the actin cytoskeleton induced by extracellular signals such as growth factors. In mammalian cells, Cdc42 regulates the formation of filopodia, whereas Rac regulates lamellipodia formation and membrane ruffling, and RhoA regulates the formation of stress fibers. Recently, the serine/threonine protein kinase p65pak autophosphorylates, thereby increasing its catalytic activity towards exogenous substrates. This kinase is therefore a candidate effector for the changes in cell shape induced by growth factors. Results: Here, we report that the microinjection of activated Pak1 protein into quiescent Swiss 3T3 cells induces the rapid formation of polarized filopodia and membrane ruffles. The prolonged overexpression of Pak1 amino-terminal mutants that are unable to bind Cdc42 or Rac1 results in the accumulation of filamentous actin in large, polarized membrane ruffles and the formation of vinculin-containing focal complexes within these structures. This phenotype resembles that seen in motile fibroblasts. The amino-terminal Pak1 mutant displays enhanced binding to the adaptor protein Nck, which contains three Src-homology 3 (SH3) domains. Mutation of a proline residue within a conserved SH3-binding region at the amino terminus of Pak1 interferes with SH3-protein binding and alters the effects of Pak1 on the cytoskeleton. Conclusions: These results indicate that Pak1, acting through a protein that contains an SH3 domain, regulates the structure of the actin cytoskeleton in mammalian cells, and may serve as an effector for Cdc42 and/or Rac1 in promoting cell motility.