PMID- 10209029
OWN - NLM
STAT- MEDLINE
DCOM- 19990517
LR  - 20190508
IS  - 0021-9525 (Print)
IS  - 0021-9525 (Linking)
VI  - 145
IP  - 2
DP  - 1999 Apr 19
TI  - Phosphorylation of adducin by Rho-kinase plays a crucial role in cell motility.
PG  - 347-61
AB  - 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.
FAU - Fukata, Y
AU  - Fukata Y
AD  - Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 
      630-0101, Japan.
FAU - Oshiro, N
AU  - Oshiro N
FAU - Kinoshita, N
AU  - Kinoshita N
FAU - Kawano, Y
AU  - Kawano Y
FAU - Matsuoka, Y
AU  - Matsuoka Y
FAU - Bennett, V
AU  - Bennett V
FAU - Matsuura, Y
AU  - Matsuura Y
FAU - Kaibuchi, K
AU  - Kaibuchi K
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Cell Biol
JT  - The Journal of cell biology
JID - 0375356
RN  - 0 (Calmodulin-Binding Proteins)
RN  - 0 (Cytoskeletal Proteins)
RN  - 0 (GTPase-Activating Proteins)
RN  - 0 (Intracellular Signaling Peptides and Proteins)
RN  - 0 (Oligopeptides)
RN  - 0 (Peptide Fragments)
RN  - 0 (Recombinant Proteins)
RN  - 0 (adducin)
RN  - 0 (rho GTPase-activating protein)
RN  - 2ZD004190S (Threonine)
RN  - 67256-21-7 (Hepatocyte Growth Factor)
RN  - EC 2.7.11.1 (Protein-Serine-Threonine Kinases)
RN  - EC 2.7.11.1 (rho-Associated Kinases)
RN  - EC 3.6.1.- (GTP-Binding Proteins)
RN  - NI40JAQ945 (Tetradecanoylphorbol Acetate)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Calmodulin-Binding Proteins/chemistry/*metabolism
MH  - Cell Line
MH  - Cell Membrane/drug effects/physiology/ultrastructure
MH  - Cell Movement
MH  - Cytoskeletal Proteins/metabolism
MH  - Cytoskeleton/drug effects/*physiology/ultrastructure
MH  - Dogs
MH  - GTP-Binding Proteins/metabolism
MH  - *GTPase-Activating Proteins
MH  - Hepatocyte Growth Factor/pharmacology
MH  - Humans
MH  - Intracellular Signaling Peptides and Proteins
MH  - Kidney
MH  - Models, Biological
MH  - Molecular Sequence Data
MH  - Oligopeptides/chemical synthesis/chemistry/metabolism
MH  - Peptide Fragments/chemistry/isolation & purification
MH  - Phosphorylation
MH  - Protein-Serine-Threonine Kinases/*metabolism
MH  - Recombinant Proteins/chemistry/metabolism
MH  - Tetradecanoylphorbol Acetate/pharmacology
MH  - Threonine
MH  - rho-Associated Kinases
PMC - PMC2133101
EDAT- 1999/04/20 00:00
MHDA- 1999/04/20 00:01
CRDT- 1999/04/20 00:00
PHST- 1999/04/20 00:00 [pubmed]
PHST- 1999/04/20 00:01 [medline]
PHST- 1999/04/20 00:00 [entrez]
AID - 10.1083/jcb.145.2.347 [doi]
PST - ppublish
SO  - J Cell Biol. 1999 Apr 19;145(2):347-61. doi: 10.1083/jcb.145.2.347.