Myosin II transport, organization, and phosphorylation: evidence for cortical flow/solation-contraction coupling during cytokinesis and cell locomotion

RL DeBiasio, GM LaRocca, PL Post and DL Taylor

Division of Molecular Sciences, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213, USA.

The mechanism of cytokinesis has been difficult to define because of the short duration and the temporal-spatial dynamics involved in the formation, activation, force production, and disappearance of the cleavage furrow. We have investigated the structural and chemical dynamics of myosin II in living Swiss 3T3 cells from prometaphase through the separation and migration of daughter cells. The structural and chemical dynamics of myosin II have been defined using the semiautomated, multimode light microscope, together with a fluorescent analogue of myosin II and a fluorescent biosensor of myosin II regulatory light chain (RLC) phosphorylation at serine 19. The correlation of image data from live cells using different modes of light microscopy allowed interpretations not possible from single-mode investigations. Myosin II transported toward the equatorial plane from adjacent regions, forming three-dimensional fibers that spanned the volume of the equator during anaphase and telophase. A global phosphorylation of myosin II at serine 19 of the RLC was initiated at anaphase when cortical myosin II transport started. The phosphorylation of myosin II remained high near the equatorial plane through telophase and into cytokinesis, whereas the phosphorylation of myosin II at serine 19 of the RLC decreased at the poles. The timing and pattern of phosphorylation was the same as the shortening of myosin II-based fibers in the cleavage furrow. Myosin II-based fibers shortened and transported out of the cleavage furrow into the tails of the two daughter cells late in cytokinesis. The patterns of myosin II transport, phosphorylation, and shortening of fibers in the migrating daughter cells were similar to that previously defined for cells migrating in a wound in vitro. The temporal-spatial patterns and dynamics of myosin II transport, phosphorylation at serine 19 of the RLC, and the shortening and disappearance of myosin II-based fibers support the proposal that a combination of the cortical flow hypothesis and the solation-contraction coupling hypothesis explain key aspects of cytokinesis and polarized cell locomotion.

Volume 7, Issue 8, pp. 1259-1282, 08/01/1996
Copyright © 1996 by The American Society for Cell Biology

This article has been cited by other articles:

				S. R. Wylie and P. D. Chantler Myosin IIC: A Third Molecular Motor Driving Neuronal Dynamics Mol. Biol. Cell, September 1, 2008; 19(9): 3956 - 3968. [Abstract] [Full Text] [PDF]

				K. Murthy and P. Wadsworth Dual role for microtubules in regulating cortical contractility during cytokinesis J. Cell Sci., July 15, 2008; 121(14): 2350 - 2359. [Abstract] [Full Text] [PDF]

				G. R.X. Hickson and P. H. O'Farrell Rho-dependent control of anillin behavior during cytokinesis J. Cell Biol., January 28, 2008; 180(2): 285 - 294. [Abstract] [Full Text] [PDF]

				M. Zhou and Y.-L. Wang Distinct Pathways for the Early Recruitment of Myosin II and Actin to the Cytokinetic Furrow Mol. Biol. Cell, January 1, 2008; 19(1): 318 - 326. [Abstract] [Full Text] [PDF]

				E. Paluch, J. van der Gucht, and C. Sykes Cracking up: symmetry breaking in cellular systems J. Cell Biol., December 4, 2006; 175(5): 687 - 692. [Abstract] [Full Text] [PDF]

				S. S. Jana, S. Kawamoto, and R. S. Adelstein A Specific Isoform of Nonmuscle Myosin II-C Is Required for Cytokinesis in a Tumor Cell Line J. Biol. Chem., August 25, 2006; 281(34): 24662 - 24670. [Abstract] [Full Text] [PDF]

				D. Saul, L. Fabian, A. Forer, and J. A. Brill Continuous phosphatidylinositol metabolism is required for cleavage of crane fly spermatocytes J. Cell Sci., August 1, 2004; 117(17): 3887 - 3896. [Abstract] [Full Text] [PDF]

				N. G. Dulyaninova, Y. V. Patskovsky, and A. R. Bresnick The N-terminus of the long MLCK induces a disruption in normal spindle morphology and metaphase arrest J. Cell Sci., March 15, 2004; 117(8): 1481 - 1493. [Abstract] [Full Text] [PDF]

				T. Chitapanarux, S. L. Chen, H. Lee, A. C. Melton, and H. F. Yee Jr. C-type natriuretic peptide induces human colonic myofibroblast relaxation Am J Physiol Gastrointest Liver Physiol, January 1, 2004; 286(1): G31 - G36. [Abstract] [Full Text] [PDF]

				H.-H. Kong and T. D. Pollard Intracellular localization and dynamics of myosin-II and myosin-IC in live Acanthamoeba by transient transfection of EGFP fusion proteins J. Cell Sci., March 14, 2003; 115(24): 4993 - 5002. [Abstract] [Full Text] [PDF]

				A. Royou, W. Sullivan, and R. Karess Cortical recruitment of nonmuscle myosin II in early syncytial Drosophila embryos: its role in nuclear axial expansion and its regulation by Cdc2 activity J. Cell Biol., July 8, 2002; 158(1): 127 - 137. [Abstract] [Full Text] [PDF]

				A. J. Piekny and P. E. Mains Rho-binding kinase (LET-502) and myosin phosphatase (MEL-11) regulate cytokinesis in the early Caenorhabditis elegans embryo J. Cell Sci., January 6, 2002; 115(11): 2271 - 2282. [Abstract] [Full Text] [PDF]

				S. Yumura Myosin II dynamics and cortical flow during contractile ring formation in Dictyostelium cells J. Cell Biol., July 9, 2001; 154(1): 137 - 146. [Abstract] [Full Text] [PDF]

				Poperechnaya, Varlamova, Lin, J. Stull, and A. Bresnick Localization and Activity of Myosin Light Chain Kinase Isoforms during the Cell Cycle J. Cell Biol., October 30, 2000; 151(3): 697 - 708. [Abstract] [Full Text] [PDF]

				R. Fukui, M. Amakawa, M. Hoshiga, N. Shibata, E. Kohbayashi, M. Seto, Y. Sasaki, T. Ueno, N. Negoro, T. Nakakoji, et al. Increased migration in late G1 phase in cultured smooth muscle cells Am J Physiol Cell Physiol, October 1, 2000; 279(4): C999 - C1007. [Abstract] [Full Text] [PDF]

				H. A. Benink, C. A. Mandato, and W. M. Bement Analysis of Cortical Flow Models In Vivo Mol. Biol. Cell, August 1, 2000; 11(8): 2553 - 2563. [Abstract] [Full Text]

				F Motegi, K Nakano, and I Mabuchi Molecular mechanism of myosin-II assembly at the division site in Schizosaccharomyces pombe J. Cell Sci., January 5, 2000; 113(10): 1813 - 1825. [Abstract] [PDF]

				K. Burton, J. H. Park, and D. L. Taylor Keratocytes Generate Traction Forces in Two Phases Mol. Biol. Cell, November 1, 1999; 10(11): 3745 - 3769. [Abstract] [Full Text]

				C. B. Shuster and D. R. Burgess Parameters that Specify the Timing of Cytokinesis J. Cell Biol., September 6, 1999; 146(5): 981 - 992. [Abstract] [Full Text] [PDF]

				M. A. Sells, J. T. Boyd, and J. Chernoff p21-Activated Kinase 1 (Pak1) Regulates Cell Motility in Mammalian Fibroblasts J. Cell Biol., May 17, 1999; 145(4): 837 - 849. [Abstract] [Full Text] [PDF]

				G. Totsukawa, Y. Yamakita, S. Yamashiro, H. Hosoya, D. J. Hartshorne, and F. Matsumura Activation of Myosin Phosphatase Targeting Subunit by Mitosis-specific Phosphorylation J. Cell Biol., February 22, 1999; 144(4): 735 - 744. [Abstract] [Full Text] [PDF]

				S Shu, R. Lee, J. LeBlanc-Straceski, and T. Uyeda Role of myosin II tail sequences in its function and localization at the cleavage furrow in Dictyostelium J. Cell Sci., January 7, 1999; 112(13): 2195 - 2201. [Abstract] [PDF]

				L. A. Silveira, J. L. Smith, J. L. Tan, and J. A. Spudich MLCK-A, an unconventional myosin light chain kinase from Dictyostelium, is activated by a cGMP-dependent pathway PNAS, October 27, 1998; 95(22): 13000 - 13005. [Abstract] [Full Text] [PDF]

				F. Matsumura, S. Ono, Y. Yamakita, G. Totsukawa, and S. Yamashiro Specific Localization of Serine 19 Phosphorylated Myosin II during Cell Locomotion and Mitosis of Cultured Cells J. Cell Biol., January 12, 1998; 140(1): 119 - 129. [Abstract] [Full Text] [PDF]

				R. Neujahr, C. Heizer, R. Albrecht, M. Ecke, J.-M. Schwartz, I. Weber, and G. Gerisch Three-dimensional Patterns and Redistribution of Myosin II and Actin in Mitotic Dictyostelium Cells J. Cell Biol., December 29, 1997; 139(7): 1793 - 1804. [Abstract] [Full Text] [PDF]

				J.-H. Zang, G. Cavet, J. H. Sabry, P. Wagner, S. L. Moores, and J. A. Spudich On the Role of Myosin-II in Cytokinesis: Division of Dictyostelium Cells under Adhesive and Nonadhesive Conditions Mol. Biol. Cell, December 1, 1997; 8(12): 2617 - 2629. [Abstract] [Full Text]

				K. A. Giuliano, R. L. DeBiasio, R. T. Dunlay, A. Gough, J. M. Volosky, J. Zock, G. N. Pavlakis, and D. L. Taylor High-Content Screening: A New Approach to Easing Key Bottlenecks in the Drug Discovery Process J Biomol Screen, June 1, 1997; 2(4): 249 - 259. [Abstract] [PDF]

				B. F. Schmidt, J. Chao, Z. Zhu, R. L. DeBiasio, and G. Fisher Signal Amplification in the Detection of Single-copy DNA and RNA by Enzyme-catalyzed Deposition (CARD) of the Novel Fluorescent Reporter Substrate Cy3.29-Tyramide J. Histochem. Cytochem., March 1, 1997; 45(3): 365 - 374. [Abstract] [Full Text] [PDF]

				R Neujahr, C Heizer, and G Gerisch Myosin II-independent processes in mitotic cells of Dictyostelium discoideum: redistribution of the nuclei, re-arrangement of the actin system and formation of the cleavage furrow J. Cell Sci., January 1, 1997; 110(2): 123 - 137. [Abstract] [PDF]

				C. A. Mandato and W. M. Bement Contraction and polymerization cooperate to assemble and close actomyosin rings around Xenopus oocyte wounds J. Cell Biol., August 20, 2001; 154(4): 785 - 798. [Abstract] [Full Text] [PDF]