In vivo phosphorylation of regulatory light chain of myosin II during mitosis of cultured cells

doi:10.1083/jcb.124.1.129

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In vivo phosphorylation of regulatory light chain of myosin II during mitosis of cultured cells

Y Yamakita, S Yamashiro and F Matsumura
Department of Molecular Biology and Biochemistry, Rutgers University, Piscataway, New Jersey 08855-1059.

Phosphorylation of the regulatory light chain of myosin II (MLC) controls the contractility of actomyosin in nonmuscle and muscle cells. It has been reported that cdc2 phosphorylates MLC in vitro at Ser-1 or Ser-2 and Thr-9 which protein kinase C phosphorylates (Satterwhite, L. L., M. J. Lohka, K. L. Wilson, T. Y. Scherson, L. K. Cisek, J. L. Corden, and T. D. Pollard. 1992 J. Cell Biol. 118:595-605). We have examined in vivo phosphorylation of MLC during mitosis and after the release of mitotic arrest. Phosphate incorporation of MLC in mitotic cells is found to be 6-12 times greater than that in nonmitotic cells. Phosphopeptide maps have revealed that the MLC from mitotic cells is phosphorylated at Ser-1 and/or Ser-2 (Ser-1/2), but not at Thr-9. MLC is also phosphorylated to a much lesser extent at Ser-19 which myosin light chain kinase phosphorylates. On the other hand, MLC of nonmitotic cells is phosphorylated at Ser-19 but not at Ser-1/2. The extent of phosphate incorporation is doubled at 30 min after the release of mitotic arrest when some cells start cytokinesis. Phosphopeptide analyses have revealed that the phosphorylation at Ser-19 is increased 20 times, while the phosphorylation at Ser-1/2 is decreased by half. This high extent of MLC phosphorylation at Ser-19 is maintained for another 30 min and gradually decreased to near the level of interphase cells as cells complete spreading at 180 min. On the other hand, phosphorylation at Ser-1/2 is decreased to 18% at 60 min, and is practically undetectable at 180 min after the release of mitotic arrest. The stoichiometry of MLC phosphorylation has been determined by quantitation of phosphorylated and unphosphorylated forms of MLC separated on 2D gels. The molar ratio of phosphorylated MLC to total MLC is found to be 0.16 +/- 0.06 and 0.31 +/- 0.05 in interphase and mitotic cells, respectively. The ratio is increased to 0.49 +/- 0.05 at 30 min after the release of mitotic arrest. These results suggest that the change in the phosphorylation site from Ser-1/2 to Ser-19 plays an important role in signaling cytokinesis.
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Asiedu, M., Wu, D., Matsumura, F., Wei, Q. (2009). Centrosome/Spindle Pole-associated Protein Regulates Cytokinesis via Promoting the Recruitment of MyoGEF to the Central Spindle. Mol. Biol. Cell 20: 1428-1440 [Abstract] [Full Text]
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Webb, S. E, Li, W. M., Miller, A. L (2008). Calcium signalling during the cleavage period of zebrafish development. Phil Trans R Soc B 363: 1363-1369 [Abstract] [Full Text]
Komatsu, S., Ikebe, M. (2007). The Phosphorylation of Myosin II at the Ser1 and Ser2 Is Critical for Normal Platelet-derived Growth Factor induced Reorganization of Myosin Filaments. Mol. Biol. Cell 18: 5081-5090 [Abstract] [Full Text]
Han, Y.-J., Hu, W.-Y., Chernaya, O., Antic, N., Gu, L., Gupta, M., Piano, M., de Lanerolle, P. (2006). Increased Myosin Light Chain Kinase Expression in Hypertension: Regulation by Serum Response Factor via an Insertion Mutation in the Promoter. Mol. Biol. Cell 17: 4039-4050 [Abstract] [Full Text]
Zhao, W.-m., Fang, G. (2005). Anillin Is a Substrate of Anaphase-promoting Complex/Cyclosome (APC/C) That Controls Spatial Contractility of Myosin during Late Cytokinesis. J. Biol. Chem. 280: 33516-33524 [Abstract] [Full Text]
Zhao, W.-m., Fang, G. (2005). MgcRacGAP controls the assembly of the contractile ring and the initiation of cytokinesis. Proc. Natl. Acad. Sci. USA 102: 13158-13163 [Abstract] [Full Text]
Fazal, F., Gu, L., Ihnatovych, I., Han, Y., Hu, W., Antic, N., Carreira, F., Blomquist, J. F., Hope, T. J., Ucker, D. S., de Lanerolle, P. (2005). Inhibiting Myosin Light Chain Kinase Induces Apoptosis In Vitro and In Vivo. Mol. Cell. Biol. 25: 6259-6266 [Abstract] [Full Text]
Straight, A. F., Field, C. M., Mitchison, T. J. (2005). Anillin Binds Nonmuscle Myosin II and Regulates the Contractile Ring. Mol. Biol. Cell 16: 193-201 [Abstract] [Full Text]
Dulyaninova, N. G., Patskovsky, Y. V., Bresnick, A. R. (2004). The N-terminus of the long MLCK induces a disruption in normal spindle morphology and metaphase arrest. J. Cell Sci. 117: 1481-1493 [Abstract] [Full Text]
Yamashiro, S., Totsukawa, G., Yamakita, Y., Sasaki, Y., Madaule, P., Ishizaki, T., Narumiya, S., Matsumura, F. (2003). Citron Kinase, a Rho-dependent Kinase, Induces Di-phosphorylation of Regulatory Light Chain of Myosin II. Mol. Biol. Cell 14: 1745-1756 [Abstract] [Full Text]
Maddox, A. S., Burridge, K. (2003). RhoA is required for cortical retraction and rigidity during mitotic cell rounding. JCB 160: 255-265 [Abstract] [Full Text]
Royou, A., Sullivan, W., Karess, R. (2002). Cortical recruitment of nonmuscle myosin II in early syncytial Drosophila embryos: its role in nuclear axial expansion and its regulation by Cdc2 activity. JCB 158: 127-137 [Abstract] [Full Text]
Guertin, D. A., Trautmann, S., McCollum, D. (2002). Cytokinesis in Eukaryotes. Microbiol. Mol. Biol. Rev. 66: 155-178 [Abstract] [Full Text]
Noguchi, T., Mabuchi, I. (2002). Localized Calcium Signals along the Cleavage Furrow of the Xenopus Egg Are Not Involved in Cytokinesis. Mol. Biol. Cell 13: 1263-1273 [Abstract] [Full Text]
Poperechnaya, A., Varlamova, O., Lin, P.-j., Stull, J. T., Bresnick, A. R. (2000). Localization and Activity of Myosin Light Chain Kinase Isoforms during the Cell Cycle. JCB 151: 697-708 [Abstract] [Full Text]
Wei, Q., Adelstein, R. S. (2000). Conditional Expression of a Truncated Fragment of Nonmuscle Myosin II-A Alters Cell Shape but Not Cytokinesis in HeLa Cells. Mol. Biol. Cell 11: 3617-3627 [Abstract] [Full Text]
Totsukawa, G., Yamakita, Y., Yamashiro, S., Hartshorne, D. J., Sasaki, Y., Matsumura, F. (2000). Distinct Roles of Rock (Rho-Kinase) and Mlck in Spatial Regulation of Mlc Phosphorylation for Assembly of Stress Fibers and Focal Adhesions in 3t3 Fibroblasts. JCB 150: 797-806 [Abstract] [Full Text]
Balasubramanian, M., McCollum, D, Surana, U (2000). Tying the knot: linking cytokinesis to the nuclear cycle. J. Cell Sci. 113: 1503-1513 [Abstract]
Motegi, F, Nakano, K, Mabuchi, I (2000). Molecular mechanism of myosin-II assembly at the division site in Schizosaccharomyces pombe. J. Cell Sci. 113: 1813-1825 [Abstract]
Foe, V., Field, C., Odell, G. (2000). Microtubules and mitotic cycle phase modulate spatiotemporal distributions of F-actin and myosin II in Drosophila syncytial blastoderm embryos. Development 127: 1767-1787 [Abstract]
Shuster, C. B., Burgess, D. R. (1999). Parameters That Specify the Timing of Cytokinesis. JCB 146: 981-992 [Abstract] [Full Text]
Turner, J. R., Angle, J. M., Black, E. D., Joyal, J. L., Sacks, D. B., Madara, J. L. (1999). PKC-dependent regulation of transepithelial resistance: roles of MLC and MLC kinase. Am. J. Physiol. Cell Physiol. 277: C554-C562 [Abstract] [Full Text]
McCollum, D., Feoktistova, A., Gould, K. L. (1999). Phosphorylation of the Myosin-II Light Chain Does Not Regulate the Timing of Cytokinesis in Fission Yeast. J. Biol. Chem. 274: 17691-17695 [Abstract] [Full Text]
Sanders, L. C., Matsumura, F., Bokoch, G. M., de Lanerolle, P. (1999). Inhibition of Myosin Light Chain Kinase by p21-Activated Kinase. Science 283: 2083-2085 [Abstract] [Full Text]
Totsukawa, G., Yamakita, Y., Yamashiro, S., Hosoya, H., Hartshorne, D. J., Matsumura, F. (1999). Activation of Myosin Phosphatase Targeting Subunit by Mitosis-specific Phosphorylation. JCB 144: 735-744 [Abstract] [Full Text]
Shannon, K. B., Li, R. (1999). The Multiple Roles of Cyk1p in the Assembly and Function of the Actomyosin Ring in Budding Yeast. Mol. Biol. Cell 10: 283-296 [Abstract] [Full Text]
Li, C., Heim, R, Lu, P, Pu, Y, Tsien, R., Chang, D. (1999). Dynamic redistribution of calmodulin in HeLa cells during cell division as revealed by a GFP-calmodulin fusion protein technique. J. Cell Sci. 112: 1567-1577 [Abstract]
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]
Wu, Y., Dowbenko, D., Lasky, L. A. (1998). PSTPIP 2,�a Second Tyrosine Phosphorylated, Cytoskeletal-associated Protein That Binds a PEST-type Protein-tyrosine Phosphatase. J. Biol. Chem. 273: 30487-30496 [Abstract] [Full Text]
Silveira, L. A., Smith, J. L., Tan, J. L., Spudich, J. A. (1998). MLCK-A, an unconventional myosin light chain kinase from Dictyostelium, is activated by a cGMP-dependent pathway. Proc. Natl. Acad. Sci. USA 95: 13000-13005 [Abstract] [Full Text]
Simerly, C., Nowak, G., de Lanerolle, P., Schatten, G. (1998). Differential Expression and Functions of Cortical Myosin IIA and IIB Isotypes during Meiotic Maturation, Fertilization, and Mitosis in Mouse Oocytes and Embryos. Mol. Biol. Cell 9: 2509-2525 [Abstract] [Full Text]
Lippincott, J., Li, R. (1998). Sequential Assembly of Myosin II, an IQGAP-like Protein, and Filamentous Actin to a Ring Structure Involved in Budding Yeast Cytokinesis. JCB 140: 355-366 [Abstract] [Full Text]
Matsumura, F., Ono, S., Yamakita, Y., Totsukawa, G., Yamashiro, S. (1998). Specific Localization of Serine 19 Phosphorylated Myosin II during Cell Locomotion and Mitosis of Cultured Cells. JCB 140: 119-129 [Abstract] [Full Text]
Sohrmann, M., Schmidt, S., Hagan, I., Simanis, V. (1998). Asymmetric segregation on spindle poles of the Schizosaccharomyces pombe septum-inducing protein kinase�Cdc7p. Genes Dev. 12: 84-94 [Abstract] [Full Text]
Jordan, P., Karess, R. (1997). Myosin Light Chain-activating Phosphorylation Sites Are Required for Oogenesis in Drosophila. JCB 139: 1805-1819 [Abstract] [Full Text]
Zang, J.-H., Cavet, G., Sabry, J. H., Wagner, P., Moores, S. L., Spudich, J. A. (1997). On the Role of Myosin-II in Cytokinesis: Division of Dictyostelium Cells under Adhesive and Nonadhesive Conditions. Mol. Biol. Cell 8: 2617-2629 [Abstract] [Full Text]
Gould, K. L., Simanis, V. (1997). The control of septum formation in fission�yeast. Genes Dev. 11: 2939-2951 [Full Text]
Spencer, S., Dowbenko, D., Cheng, J., Li, W., Brush, J., Utzig, S., Simanis, V., Lasky, L. A. (1997). PSTPIP: A Tyrosine Phosphorylated Cleavage Furrow-associated Protein that Is a Substrate for a PEST Tyrosine Phosphatase. JCB 138: 845-860 [Abstract] [Full Text]
Walker, G., Shuster, C., Burgess, D. (1997). Microtubule-entrained kinase activities associated with the cortical cytoskeleton during cytokinesis. J. Cell Sci. 110: 1373-1386 [Abstract]
Holmes, JenniferK., Solomon, MarkJ. (1996). A Predictive Scale for Evaluating Cyclin-dependent Kinase Substrates. A COMPARISON OF p34cdc2 AND p33cdk2. J. Biol. Chem. 271: 25240-25246 [Abstract] [Full Text]
Yamakita, Y., Ono, S., Matsumura, F., Yamashiro, S. (1996). Phosphorylation of Human Fascin Inhibits Its Actin Binding and Bundling Activities. J. Biol. Chem. 271: 12632-12638 [Abstract] [Full Text]
Kelley, C. A., Oberman, F., Yisraeli, J. K., Adelstein, R. S. (1995). A Xenopus Nonmuscle Myosin Heavy Chain Isoform Is Phosphorylated by Cyclin-p34[IMAGE] Kinase during Meiosis. J. Biol. Chem. 270: 1395-1401 [Abstract] [Full Text]
Mabuchi, I. (1994). Cleavage furrow: timing of emergence of contractile ring actin filaments and establishment of the contractile ring by filament bundling in sea urchin eggs. J. Cell Sci. 107: 1853-1862 [Abstract]
Komatsu, S., Yano, T., Shibata, M., Tuft, R. A., Ikebe, M. (2000). Effects of the Regulatory Light Chain Phosphorylation of Myosin II on Mitosis and Cytokinesis of Mammalian Cells. J. Biol. Chem. 275: 34512-34520 [Abstract] [Full Text]