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PMID: 23770677 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

In vitro contraction of cytokinetic ring depends on myosin II but not on actin dynamics.

Nature cell biology ·Vol. 15 ·No. 7 ·2013-07-00 ·Pages 853-9

Mishra M, Kashiwazaki J, Takagi T, Srinivasan R, Huang Y, Balasubramanian MK, Mabuchi I

Abstract

Cytokinesis in many eukaryotes involves the contraction of an actomyosin-based contractile ring. However, the detailed mechanism of contractile ring contraction is not fully understood. Here, we establish an experimental system to study contraction of the ring to completion in vitro. We show that the contractile ring of permeabilized fission yeast cells undergoes rapid contraction in an ATP- and myosin-II-dependent manner in the absence of other cytoplasmic constituents. Surprisingly, neither actin polymerization nor its disassembly is required for contraction of the contractile ring, although addition of exogenous actin-crosslinking proteins blocks ring contraction. Using contractile rings generated from fission yeast cytokinesis mutants, we show that not all proteins required for assembly of the ring are required for its contraction in vitro. Our work provides the beginnings of the definition of a minimal contraction-competent cytokinetic ring apparatus.

MeSH Terms
Actins/metabolism Adenosine Triphosphate/metabolism Cell Division Cell Membrane/metabolism Cytokinesis/physiology Green Fluorescent Proteins/metabolism Myosin Type II/metabolism Schizosaccharomyces/cytology,metabolism Schizosaccharomyces pombe Proteins/metabolism
Chemicals
Actins Schizosaccharomyces pombe Proteins Green Fluorescent Proteins Adenosine Triphosphate Myosin Type II
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Mishra Mithilesh
Temasek Life Sciences Laboratory, The National University of Singapore, 1 Research Link, Singapore 117604, Singapore.
Kashiwazaki Jun
Takagi Tomoko
Srinivasan Ramanujam
Huang Yinyi
Balasubramanian Mohan K
Mabuchi Issei
References (39)
39 references, click to expand
  1. Identification of Myo3, a second type-II myosin heavy chain in the fission yeast Schizosaccharomyces pombe.
    FEBS Lett. 1997 Dec 29;420(2-3):161-6 PMID: 9459302
  2. Heterologous modules for efficient and versatile PCR-based gene targeting in Schizosaccharomyces pombe.
    Yeast. 1998 Jul;14(10):943-51 PMID: 9717240
  3. UCS protein Rng3p activates actin filament gliding by fission yeast myosin-II.
    J Cell Biol. 2004 Oct 25;167(2):315-25 PMID: 15504913
  4. Actin dynamics in the contractile ring during cytokinesis in fission yeast.
    Nature. 2002 Sep 5;419(6902):82-6 PMID: 12214236
  5. Biochemical aspects of cytokinesis.
    Int Rev Cytol. 1986;101:175-213 PMID: 3516914
  6. Actin depolymerization drives actomyosin ring contraction during budding yeast cytokinesis.
    Dev Cell. 2012 Jun 12;22(6):1247-60 PMID: 22698284
  7. Tropomyosin and myosin-II cellular levels promote actomyosin ring assembly in fission yeast.
    Mol Biol Cell. 2010 Mar 15;21(6):989-1000 PMID: 20110347
  8. Identification of a second myosin-II in Schizosaccharomyces pombe: Myp2p is conditionally required for cytokinesis.
    Mol Biol Cell. 1997 Dec;8(12):2693-705 PMID: 9398685
  9. Synthesis of biotinylated xestoquinone that retains inhibitory activity against Ca2+ ATPase of skeletal muscle myosin.
    Bioorg Med Chem. 2003 Jul 17;11(14):3077-82 PMID: 12818670
  10. A new tropomyosin essential for cytokinesis in the fission yeast S. pombe.
    Nature. 1992 Nov 5;360(6399):84-7 PMID: 1436080
  11. Fission yeast IQGAP arranges actin filaments into the cytokinetic contractile ring.
    EMBO J. 2009 Oct 21;28(20):3117-31 PMID: 19713940
  12. Type II myosin heavy chain encoded by the myo2 gene composes the contractile ring during cytokinesis in Schizosaccharomyces pombe.
    J Cell Biol. 1997 Jun 16;137(6):1309-19 PMID: 9182664
  13. The fission yeast cytokinesis formin Cdc12p is a barbed end actin filament capping protein gated by profilin.
    J Cell Biol. 2003 Jun 9;161(5):875-87 PMID: 12796476
  14. Molecular genetic analysis of fission yeast Schizosaccharomyces pombe.
    Methods Enzymol. 1991;194:795-823 PMID: 2005825
  15. Isolation and characterization of new fission yeast cytokinesis mutants.
    Genetics. 1998 Jul;149(3):1265-75 PMID: 9649519
  16. Actin-depolymerizing protein Adf1 is required for formation and maintenance of the contractile ring during cytokinesis in fission yeast.
    Mol Biol Cell. 2006 Apr;17(4):1933-45 PMID: 16467379
  17. Rng2p, a protein required for cytokinesis in fission yeast, is a component of the actomyosin ring and the spindle pole body.
    Curr Biol. 1998 May 21;8(11):611-21 PMID: 9635188
  18. Dissecting temporal and spatial control of cytokinesis with a myosin II Inhibitor.
    Science. 2003 Mar 14;299(5613):1743-7 PMID: 12637748
  19. Mechanics of cytokinesis in eukaryotes.
    Curr Opin Cell Biol. 2010 Feb;22(1):50-6 PMID: 20031383
  20. Split decisions: coordinating cytokinesis in yeast.
    Trends Cell Biol. 2005 Jan;15(1):10-8 PMID: 15653073
  21. cdc12p, a protein required for cytokinesis in fission yeast, is a component of the cell division ring and interacts with profilin.
    J Cell Biol. 1997 Apr 7;137(1):169-82 PMID: 9105045
  22. Directionality of F-actin cables changes during the fission yeast cell cycle.
    Nat Cell Biol. 2005 Sep;7(9):916-7 PMID: 16136186
  23. Dephosphorylation of F-BAR protein Cdc15 modulates its conformation and stimulates its scaffolding activity at the cell division site.
    Mol Cell. 2010 Jul 9;39(1):86-99 PMID: 20603077
  24. Isolation and partial purification of the Saccharomyces cerevisiae cytokinetic apparatus.
    Cytoskeleton (Hoboken). 2010 Jan;67(1):13-22 PMID: 19790107
  25. Interactions among a fimbrin, a capping protein, and an actin-depolymerizing factor in organization of the fission yeast actin cytoskeleton.
    Mol Biol Cell. 2001 Nov;12(11):3515-26 PMID: 11694585
  26. Schizosaccharomyces pombe cdc4+ gene encodes a novel EF-hand protein essential for cytokinesis.
    J Cell Biol. 1995 Aug;130(3):651-60 PMID: 7622565
  27. Three-dimensional arrangement of F-actin in the contractile ring of fission yeast.
    J Cell Biol. 2007 Aug 27;178(5):765-71 PMID: 17724118
  28. Understanding cytokinesis: lessons from fission yeast.
    Nat Rev Mol Cell Biol. 2010 Feb;11(2):149-55 PMID: 20094054
  29. The molecular requirements for cytokinesis.
    Science. 2005 Mar 18;307(5716):1735-9 PMID: 15774750
  30. Cleavage furrow isolated from newt eggs: contraction, organization of the actin filaments, and protein components of the furrow.
    Proc Natl Acad Sci U S A. 1988 Aug;85(16):5966-70 PMID: 3413069
  31. F-actin ring formation and the role of F-actin cables in the fission yeast Schizosaccharomyces pombe.
    J Cell Sci. 2002 Mar 1;115(Pt 5):887-98 PMID: 11870208
  32. Cylindrical cellular geometry ensures fidelity of division site placement in fission yeast.
    J Cell Sci. 2012 Aug 15;125(Pt 16):3850-7 PMID: 22505610
  33. Mechanics and regulation of cytokinesis.
    Curr Opin Cell Biol. 2004 Apr;16(2):182-8 PMID: 15196562
  34. The effect of myosin antibody on the division of starfish blastomeres.
    J Cell Biol. 1977 Jul;74(1):251-63 PMID: 141455
  35. Spatial and temporal pathway for assembly and constriction of the contractile ring in fission yeast cytokinesis.
    Dev Cell. 2003 Nov;5(5):723-34 PMID: 14602073
  36. Fission yeast Rng3p: an UCS-domain protein that mediates myosin II assembly during cytokinesis.
    J Cell Sci. 2000 Jul;113 ( Pt 13):2421-32 PMID: 10852821
  37. Actin is associated with the formation of the cell wall in reverting protoplasts of the fission yeast Schizosaccharomyces pombe.
    J Cell Sci. 1989 Dec;94 ( Pt 4):635-46 PMID: 2630560
  38. Identification of two type V myosins in fission yeast, one of which functions in polarized cell growth and moves rapidly in the cell.
    Mol Biol Cell. 2001 May;12(5):1367-80 PMID: 11359928
  39. Subcellular localization and possible function of actin, tropomyosin and actin-related protein 3 (Arp3) in the fission yeast Schizosaccharomyces pombe.
    Eur J Cell Biol. 1998 Aug;76(4):288-95 PMID: 9765059
Article Info
Journal
Nature cell biology
Abbr.
Nat Cell Biol
ISSN
1476-4679
Published
2013-07-00
Epub
2013-00-16
Pages
853-9
Language
English
Region
England
NLM ID
100890575
Subset
IM
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