© The Rockefeller University Press,
0021-9525/1998//409 $5.00
The Journal of Cell Biology, Volume 141, Number 2,
, 1998 409-418
Association of the Myosin-binding Subunit of Myosin Phosphatase and Moesin: Dual Regulation of Moesin Phosphorylation by Rho-associated Kinase and Myosin Phosphatase
Yuko Fukata*,
Kazushi Kimura*,
Noriko Oshiro*,
Hideyuki Saya
,
Yoshiharu Matsuura
, and
Kozo Kaibuchi*
* Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan;
Department of Oncology, Kumamoto University School of Medicine, Kumamoto 860-0811, Japan; and
Department of Virology II, National Institute of Infectious Diseases, Tokyo 162-0052, Japan
The small GTPase Rho is believed to regulate the actin cytoskeleton and cell adhesion through its specific targets. We previously identified the Rho targets: protein kinase N, Rho-associated kinase (Rho- kinase), and the myosin-binding subunit (MBS) of myosin phosphatase. We found that in MDCK epithelial cells, MBS accumulated at the tetradecanoylphorbol-13-acetate (TPA)-induced membrane ruffling area, where moesin, a member of the ERM (ezrin, radixin, and moesin) family, was localized. Neither membrane ruffling nor an accumulation of moesin and MBS at the free-end plasma membrane was induced when MDCK cells were stimulated with TPA after the microinjection of C3, which ADP-ribosylates and inactivates Rho. MBS was colocalized with moesin at the cell–cell contact sites in MDCK cells. We also found that moesin was coimmunoprecipitated with MBS from MDCK cells. Recombinant MBS interacted with the amino-terminal domains of moesin and ezrin. Myosin phosphatase composed of the catalytic subunit and MBS showed phosphatase activity toward moesin, which was phosphorylated by Rho-kinase. The phosphatase activity was inhibited when MBS was phosphorylated by Rho-kinase. These results suggest that MBS is recruited with moesin to the plasma membrane and that myosin phosphatase and Rho-kinase regulate the phosphorylation state of moesin downstream of Rho.
Abbreviations used in this paper: aa, amino acids; A-PMSF, (p-amidinophenyl)-methanesulfonyl fluoride; C-moesin, COOH-terminal domain of moesin; ERM, ezrin, radixin, and moesin; GDI, GDP dissociation inhibitor; GST, glutathione-S-transferase; HA, hemagglutinin; MBS, myosin-binding subunit; MLC, myosin light chain; 4,5-PIP2, phosphatidylinositol 4,5-diphosphate; PI5-kinase, phosphatidylinositol 5-kinase; Rho-kinase, Rho-associated kinase; TPA, tetradecanoylphorbol-13-acetate.
This investigation was supported by grants-in-aid for Scientific Research and for Cancer Research from the Ministry of Education, Science, and Culture, Japan (1997) and by grants from the Mitsubishi Foundation and Kirin Brewery Company Limited.
Address all correspondence to Kozo Kaibuchi, M.D. and Ph.D., Division of Signal Transduction, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma 630-0101, Japan, Tel.: 81-743-72-5440. Fax: 81-743-72-5449. E-mail: kaibuchi{at}bs.aist-nara.ac.jp

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-
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-
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[Abstract]
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-
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-
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-
Gautreau, A., Louvard, D., Arpin, M.
(2000). Morphogenic Effects of Ezrin Require a Phosphorylation-Induced Transition from Oligomers to Monomers at the Plasma Membrane. JCB
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[Abstract]
[Full Text]
-
Terry-Lorenzo, R. T., Inoue, M., Connor, J. H., Haystead, T. A. J., Armbruster, B. N., Gupta, R. P., Oliver, C. J., Shenolikar, S.
(2000). Neurofilament-L Is a Protein Phosphatase-1-binding Protein Associated with Neuronal Plasma Membrane and Post-synaptic Density. J. Biol. Chem.
275: 2439-2446
[Abstract]
[Full Text]
-
Feng, J., Ito, M., Ichikawa, K., Isaka, N., Nishikawa, M., Hartshorne, D. J., Nakano, T.
(1999). Inhibitory Phosphorylation Site for Rho-associated Kinase on Smooth Muscle Myosin Phosphatase. J. Biol. Chem.
274: 37385-37390
[Abstract]
[Full Text]
-
Tsukita, S., Yonemura, S.
(1999). Cortical Actin Organization: Lessons from ERM (Ezrin/Radixin/Moesin) Proteins. J. Biol. Chem.
274: 34507-34510
[Full Text]
-
Inada, H., Togashi, H., Nakamura, Y., Kaibuchi, K., Nagata, K.-i., Inagaki, M.
(1999). Balance between Activities of Rho Kinase and Type 1 Protein Phosphatase Modulates Turnover of Phosphorylation and Dynamics of Desmin/Vimentin Filaments. J. Biol. Chem.
274: 34932-34939
[Abstract]
[Full Text]
-
Kawano, Y., Fukata, Y., Oshiro, N., Amano, M., Nakamura, T., Ito, M., Matsumura, F., Inagaki, M., Kaibuchi, K.
(1999). Phosphorylation of Myosin-Binding Subunit (Mbs) of Myosin Phosphatase by Rho-Kinase in Vivo. JCB
147: 1023-1038
[Abstract]
[Full Text]
-
Hishiya, A., Ohnishi, M., Tamura, S., Nakamura, F.
(1999). Protein Phosphatase 2C Inactivates F-actin Binding of Human Platelet Moesin. J. Biol. Chem.
274: 26705-26712
[Abstract]
[Full Text]
-
Okamoto, I., Kawano, Y., Matsumoto, M., Suga, M., Kaibuchi, K., Ando, M., Saya, H.
(1999). Regulated CD44 Cleavage under the Control of Protein Kinase C, Calcium Influx, and the Rho Family of Small G Proteins. J. Biol. Chem.
274: 25525-25534
[Abstract]
[Full Text]
-
Seasholtz, T. M., Majumdar, M., Brown, J. H.
(1999). MINIREVIEW: Rho as a Mediator of G Protein-Coupled Receptor Signaling. Mol. Pharmacol.
55: 949-956
[Full Text]
-
Huang, L., Wong, T. Y. W., Lin, R. C. C., Furthmayr, H.
(1999). Replacement of Threonine 558, a Critical Site of Phosphorylation of Moesin in Vivo, with Aspartate Activates F-actin Binding of Moesin. REGULATION BY CONFORMATIONAL CHANGE. J. Biol. Chem.
274: 12803-12810
[Abstract]
[Full Text]
-
Fukata, Y., Oshiro, N., Kinoshita, N., Kawano, Y., Matsuoka, Y., Bennett, V., Matsuura, Y., Kaibuchi, K.
(1999). Phosphorylation of Adducin by Rho-Kinase Plays a Crucial Role in Cell Motility. JCB
145: 347-361
[Abstract]
[Full Text]
-
Feng, J., Ito, M., Kureishi, Y., Ichikawa, K., Amano, M., Isaka, N., Okawa, K., Iwamatsu, A., Kaibuchi, K., Hartshorne, D. J., Nakano, T.
(1999). Rho-associated Kinase of Chicken Gizzard Smooth Muscle. J. Biol. Chem.
274: 3744-3752
[Abstract]
[Full Text]
-
Amieva, M., Litman, P, Huang, L, Ichimaru, E, Furthmayr, H
(1999). Disruption of dynamic cell surface architecture of NIH3T3 fibroblasts by the N-terminal domains of moesin and ezrin: in vivo imaging with GFP fusion proteins. J. Cell Sci.
112: 111-125
[Abstract]
-
Oshiro, N., Fukata, Y., Kaibuchi, K.
(1998). Phosphorylation of Moesin by Rho-associated Kinase (Rho-kinase) Plays a Crucial Role in the Formation of Microvilli-like Structures. J. Biol. Chem.
273: 34663-34666
[Abstract]
[Full Text]
-
Yasui, Y., Amano, M., Nagata, K.-i., Inagaki, N., Nakamura, H., Saya, H., Kaibuchi, K., Inagaki, M.
(1998). Roles of Rho-associated Kinase in Cytokinesis; Mutations in Rho-associated Kinase Phosphorylation Sites Impair Cytokinetic Segregation of Glial Filaments. JCB
143: 1249-1258
[Abstract]
[Full Text]
-
Xiao, G.-H., Beeser, A., Chernoff, J., Testa, J. R.
(2002). p21-activated Kinase Links Rac/Cdc42 Signaling to Merlin. J. Biol. Chem.
277: 883-886
[Abstract]
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-
Togashi, H., Nagata, K.-i., Takagishi, M., Saitoh, N., Inagaki, M.
(2000). Functions of a Rho-specific Guanine Nucleotide Exchange Factor in Neurite Retraction. POSSIBLE ROLE OF A PROLINE-RICH MOTIF OF KIAA0380 IN LOCALIZATION. J. Biol. Chem.
275: 29570-29578
[Abstract]
[Full Text]
-
Tan, I., Ng, C. H., Lim, L., Leung, T.
(2001). Phosphorylation of a Novel Myosin Binding Subunit of Protein Phosphatase 1 Reveals a Conserved Mechanism in the Regulation of Actin Cytoskeleton. J. Biol. Chem.
276: 21209-21216
[Abstract]
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-
Yamamoto, N., Yamamoto, S.-i., Inagaki, F., Kawaichi, M., Fukamizu, A., Kishi, N., Matsuno, K., Nakamura, K., Weinmaster, G., Okano, H., Nakafuku, M.
(2001). Role of Deltex-1 as a Transcriptional Regulator Downstream of the Notch Receptor. J. Biol. Chem.
276: 45031-45040
[Abstract]
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-
Broustas, C. G., Grammatikakis, N., Eto, M., Dent, P., Brautigan, D. L., Kasid, U.
(2002). Phosphorylation of the Myosin-binding Subunit of Myosin Phosphatase by Raf-1 and Inhibition of Phosphatase Activity. J. Biol. Chem.
277: 3053-3059
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