p160ROCK, a Rho-associated coiled-coil forming protein kinase, works downstream of Rho and induces focal adhesions.
p160ROCK is a serine/threonine protein kinase that binds selectively to GTP-Rho and is activated by this binding. To identify its function, we transfected HeLa cells with wild type and mutants of p160ROCK and examined morphology of the transfected cells. Transfection with wild type and mutants containing the kinase domain and the coiled-coil forming region induced focal adhesions and stress fibers, while no induction was observed with a kinase-defective mutant or a mutant containing only the kinase domain. Furthermore, Rho-induced formation of focal adhesions and stress fibers was inhibited by co-expression of a mutant defective in both kinase and Rho-binding activities. Rho, however, still induced an increase in F-actin content in these cells. These results suggest that p160ROCK works downstream of Rho to induce formation of focal adhesions and that Rho-induced actin polymerization is mediated by other effector(s).
Ishizaki
T
T
Department of Pharmacology, Kyoto University Faculty of Medicine, Japan.
Naito
M
M
Fujisawa
K
K
Maekawa
M
M
Watanabe
N
N
Saito
Y
Y
Narumiya
S
S
D000409:Alanine; D001483:Base Sequence; D002448:Cell Adhesion; D017931:DNA Primers; D020558:GTP Phosphohydrolases / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008239:Lysine; D018613:Microscopy, Confocal; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014162:Transfection; D054460:rho-Associated Kinases
D017931:DNA Primers; D047908:Intracellular Signaling Peptides and Proteins; D011993:Recombinant Fusion Proteins; D017346:Protein Serine-Threonine Kinases; D054460:rho-Associated Kinases; D020558:GTP Phosphohydrolases; D008239:Lysine; D000409:Alanine