PMID- 10579722
OWN - NLM
STAT- MEDLINE
DCOM- 19991230
LR  - 20190508
IS  - 0021-9525 (Print)
IS  - 0021-9525 (Linking)
VI  - 147
IP  - 5
DP  - 1999 Nov 29
TI  - Phosphorylation of myosin-binding subunit (MBS) of myosin phosphatase by
      Rho-kinase in vivo.
PG  - 1023-38
AB  - Rho-associated kinase (Rho-kinase), which is activated by the small GTPase Rho,
      phosphorylates myosin-binding subunit (MBS) of myosin phosphatase and thereby
      inactivates the phosphatase activity in vitro. Rho-kinase is thought to regulate 
      the phosphorylation state of the substrates including myosin light chain (MLC),
      ERM (ezrin/radixin/moesin) family proteins and adducin by their direct
      phosphorylation and by the inactivation of myosin phosphatase. Here we identified
      the sites of phosphorylation of MBS by Rho-kinase as Thr-697, Ser-854 and several
      residues, and prepared antibody that specifically recognized MBS phosphorylated
      at Ser-854. We found by use of this antibody that the stimulation of MDCK
      epithelial cells with tetradecanoylphorbol-13-acetate (TPA) or hepatocyte growth 
      factor (HGF) induced the phosphorylation of MBS at Ser-854 under the conditions
      in which membrane ruffling and cell migration were induced. Pretreatment of the
      cells with Botulinum C3 ADP-ribosyltransferase (C3), which is thought to
      interfere with Rho functions, or Rho-kinase inhibitors inhibited the TPA- or
      HGF-induced MBS phosphorylation. The TPA stimulation enhanced the
      immunoreactivity of phosphorylated MBS in the cytoplasm and membrane ruffling
      area of MDCK cells. In migrating MDCK cells, phosphorylated MBS as well as
      phosphorylated MLC at Ser-19 were localized in the leading edge and posterior
      region. Phosphorylated MBS was localized on actin stress fibers in REF52
      fibroblasts. The microinjection of C3 or dominant negative Rho-kinase disrupted
      stress fibers and weakened the accumulation of phosphorylated MBS in REF52 cells.
      During cytokinesis, phosphorylated MBS, MLC and ERM family proteins accumulated
      at the cleavage furrow, and the phosphorylation level of MBS at Ser-854 was
      increased. Taken together, these results indicate that MBS is phosphorylated by
      Rho-kinase downstream of Rho in vivo, and suggest that myosin phosphatase and
      Rho-kinase spatiotemporally regulate the phosphorylation state of Rho-kinase
      substrates including MLC and ERM family proteins in vivo in a cooperative manner.
FAU - Kawano, Y
AU  - Kawano Y
AD  - Division of Signal Transduction, Nara Institute of Science and Technology, 8916-5
      Takayama, Ikoma 630-0101, Japan.
FAU - Fukata, Y
AU  - Fukata Y
FAU - Oshiro, N
AU  - Oshiro N
FAU - Amano, M
AU  - Amano M
FAU - Nakamura, T
AU  - Nakamura T
FAU - Ito, M
AU  - Ito M
FAU - Matsumura, F
AU  - Matsumura F
FAU - Inagaki, M
AU  - Inagaki M
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 (Actins)
RN  - 0 (Intracellular Signaling Peptides and Proteins)
RN  - 0 (Membrane Proteins)
RN  - 0 (Myosin Light Chains)
RN  - 0 (Neurofibromin 2)
RN  - 0 (Peptides)
RN  - EC 2.7.11.1 (Protein-Serine-Threonine Kinases)
RN  - EC 2.7.11.1 (rho-Associated Kinases)
RN  - EC 3.1.3.16 (Phosphoprotein Phosphatases)
RN  - EC 3.1.3.53 (Myosin-Light-Chain Phosphatase)
RN  - EC 3.6.4.1 (Myosins)
SB  - IM
MH  - Actins/metabolism
MH  - Amino Acid Sequence
MH  - Animals
MH  - Cell Division/physiology
MH  - Cell Line
MH  - Cell Movement/physiology
MH  - Dogs
MH  - Intracellular Signaling Peptides and Proteins
MH  - Membrane Proteins/metabolism
MH  - Molecular Sequence Data
MH  - Myosin Light Chains/metabolism
MH  - Myosin-Light-Chain Phosphatase
MH  - Myosins/*metabolism
MH  - Neurofibromin 2
MH  - Peptides/metabolism
MH  - Phosphoprotein Phosphatases/analysis/*metabolism
MH  - Phosphorylation
MH  - Protein-Serine-Threonine Kinases/*metabolism
MH  - Sequence Alignment
MH  - rho-Associated Kinases
PMC - PMC2169354
EDAT- 1999/12/01 00:00
MHDA- 1999/12/01 00:01
CRDT- 1999/12/01 00:00
PHST- 1999/12/01 00:00 [pubmed]
PHST- 1999/12/01 00:01 [medline]
PHST- 1999/12/01 00:00 [entrez]
AID - 10.1083/jcb.147.5.1023 [doi]
PST - ppublish
SO  - J Cell Biol. 1999 Nov 29;147(5):1023-38. doi: 10.1083/jcb.147.5.1023.