Publications [#112878] of G Vann V. Bennett

Papers Published

1. Y Fukata, N Oshiro, N Kinoshita, Y Kawano, Y Matsuoka, V Bennett, Y Matsuura, K Kaibuchi, Phosphorylation of adducin by Rho-kinase plays a crucial role in cell motility., The Journal of cell biology, UNITED STATES, vol. 145 no. 2 (April, 1999), pp. 347-61, ISSN 0021-9525
   (last updated on 2003/02/18)

   Abstract:
   Adducin is a membrane skeletal protein that binds to actin filaments (F-actin) and thereby promotes the association of spectrin with F-actin to form a spectrin-actin meshwork beneath plasma membranes such as ruffling membranes. Rho-associated kinase (Rho- kinase), which is activated by the small guanosine triphosphatase Rho, phosphorylates alpha-adducin and thereby enhances the F-actin-binding activity of alpha-adducin in vitro. Here we identified the sites of phosphorylation of alpha-adducin by Rho-kinase as Thr445 and Thr480. We prepared antibody that specifically recognized alpha-adducin phosphorylated at Thr445, and found by use of this antibody that Rho-kinase phosphorylated alpha-adducin at Thr445 in COS7 cells in a Rho-dependent manner. Phosphorylated alpha-adducin accumulated in the membrane ruffling area of Madin-Darby canine kidney (MDCK) epithelial cells and the leading edge of scattering cells during the action of tetradecanoylphorbol-13-acetate (TPA) or hepatocyte growth factor (HGF). The microinjection of Botulinum C3 ADP-ribosyl-transferase, dominant negative Rho-kinase, or alpha-adducinT445A,T480A (substitution of Thr445 and Thr480 by Ala) inhibited the TPA-induced membrane ruffling in MDCK cells and wound-induced migration in NRK49F cells. alpha-AdducinT445D,T480D (substitution of Thr445 and Thr480 by Asp), but not alpha-adducinT445A,T480A, counteracted the inhibitory effect of the dominant negative Rho-kinase on the TPA-induced membrane ruffling in MDCK cells. Taken together, these results indicate that Rho-kinase phosphorylates alpha-adducin downstream of Rho in vivo, and that the phosphorylation of adducin by Rho-kinase plays a crucial role in the regulation of membrane ruffling and cell motility.

   Keywords:
   Amino Acid Sequence • Animals • Calmodulin-Binding Proteins • Cell Line • Cell Membrane • Cell Movement • Cytoskeletal Proteins • Cytoskeleton • Dogs • GTP-Binding Proteins • GTPase-Activating Proteins* • Hepatocyte Growth Factor • Humans • Intracellular Signaling Peptides and Proteins • Kidney • Models, Biological • Molecular Sequence Data • Oligopeptides • Peptide Fragments • Phosphorylation • Protein-Serine-Threonine Kinases • Recombinant Proteins • Tetradecanoylphorbol Acetate • Threonine • chemical synthesis • chemistry • drug effects • isolation & purification • metabolism • metabolism* • pharmacology • physiology • physiology* • ultrastructure