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Rho-kinase contributes to diphosphorylation of myosin II regulatory light chain in nonmuscle cells

Oncogene volume 21, pages 5852–5860 (2002)Cite this article

Abstract

Phosphorylation of myosin II regulatory light chain (MRLC) is important for cell motility and cytokinesis in nonmuscle cells. Although the regulation of monophosphorylated MRLC at serine 19 throughout the cell cycle was examined in detail, MRLC diphosphorylation at both threonine 18 and serine 19 is still unclear. Here we found that Rho-kinase has an activity for MRLC diphosphorylation in nonmuscle cells using sequential column chromatographies. Transfection of Rho-kinase-EGFP induced the excess diphosphorylated MRLC and the bundling of the actin filaments. Conversely, the treatment of cells with a specific inhibitor of Rho-kinase, Y-27632, resulted in the decrease of endogenous diphosphorylated MRLC and actin stress fibers. Immunolocalization studies showed that both diphosphorylated MRLC and Rho-kinase accumulated and colocalized at the contractile ring and the midbody in dividing cells. Taken together, it is suggested that Rho-kinase contributes to MRLC diphosphorylation and reorganization of actin filaments in nonmuscle cells.

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Acknowledgements

We are grateful to Welfide Corporation for providing Y-27632, Dr E Majima (Apro Science, Tokushima, Japan) for help with amino acid sequence analysis, Dr Y Yamamoto (Faculty of Applied Biological Science, Hiroshima University) for giving us chicken gizzards and Dr K Kohama (Gunma University, School of Medicine, Maebashi, Japan) for skilled advice in preparing myosin II from chicken gizzards. We also thank Mr T Iwasaki for providing GST-MRLC fusion proteins. This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan to H Hosoya.

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  1. Maki Murata-Hori

    Present address: University of Massachusetts Medical School Biotech Four, Suite 327 377 Plantation Street, Worcester, 01605, MA, USA

Authors and Affiliations

  1. Department of Biological Science, Graduate School of Science, Hiroshima University, Higashi-Hiroshima, 739-8526, Japan

    Kozue Ueda, Maki Murata-Hori & Hiroshi Hosoya

  2. Department of Molecular Radiobiology, Research Institution for Radiation Biology and Medicine, Hiroshima University, Hiroshima, 734-8553

    Masaaki Tatsuka

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  1. Kozue Ueda
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Correspondence to Hiroshi Hosoya.

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Ueda, K., Murata-Hori, M., Tatsuka, M. et al. Rho-kinase contributes to diphosphorylation of myosin II regulatory light chain in nonmuscle cells. Oncogene 21, 5852–5860 (2002). https://doi.org/10.1038/sj.onc.1205747

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