Abstract
Cytokinesis is the final stage of cell division, through which cellular constituents of mother cells are partitioned into two daughter cells resulting in the increase in cell number. In animal and fungal cells cytokinesis is mediated by an actomyosin contractile ring, which is attached to the overlying cell membrane. Contraction of this ring after chromosome segregation physically severs the mother cell into two daughters. Here we describe methods for the isolation and partial purification of the actomyosin ring from the fission yeast Schizosaccharomyces pombe and the budding yeast Saccharomyces cerevisiae, which can serve as in vitro systems to facilitate biochemical and ultrastructural analysis of cytokinesis in these genetically tractable model systems.
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References
Wu JQ, Pollard TD (2005) Counting cytokinesis proteins globally and locally in fission yeast. Science 310(5746):310–314. doi:10.1126/science.1113230
Pollard TD, Wu JQ (2010) Understanding cytokinesis: lessons from fission yeast. Nat Rev Mol Cell Biol 11(2):149–155. doi:10.1038/nrm2834
Glotzer M (2005) The molecular requirements for cytokinesis. Science 307(5716):1735–1739. doi:10.1126/science.1096896
Huang J, Huang Y, Yu H, Subramanian D, Padmanabhan A, Thadani R, Tao Y, Tang X, Wedlich-Soldner R, Balasubramanian MK (2012) Nonmedially assembled F-actin cables incorporate into the actomyosin ring in fission yeast. J Cell Biol 199(5):831–847. doi:10.1083/jcb.201209044
D’Avino PP (2009) How to scaffold the contractile ring for a safe cytokinesis - lessons from Anillin-related proteins. J Cell Sci 122(Pt 8):1071–1079. doi:10.1242/jcs.034785
van den Heuvel S (2005) The C. elegans cell cycle: overview of molecules and mechanisms. Methods Mol Biol 296:51–67
Young BA, Buser C, Drubin DG (2010) Isolation and partial purification of the Saccharomyces cerevisiae cytokinetic apparatus. Cytoskeleton 67(1):13–22. doi:10.1002/cm.20412
Mishra M, Kashiwazaki J, Takagi T, Srinivasan R, Huang Y, Balasubramanian MK, Mabuchi I (2013) In vitro contraction of cytokinetic ring depends on myosin II but not on actin dynamics. Nat Cell Biol 15(7):853–859. doi:10.1038/ncb2781
Walker GR, Kane R, Burgess DR (1994) Isolation and characterization of a sea urchin zygote cortex that supports in vitro contraction and reactivation of furrowing. J Cell Sci 107(Pt 8):2239–2248
Fujimoto H, Mabuchi I (1997) Isolation of cleavage furrows from eggs of regular sea urchins and identification of furrow-specific proteins. J Biochem 122(3):518–524
Scott JH, Schekman R (1980) Lyticase: endoglucanase and protease activities that act together in yeast cell lysis. J Bacteriol 142(2):414–423
Acknowledgments
We thank Dr. David Drubin for kindly providing the S. cerevisiae strains. This work was supported by research funds from the Wellcome Trust, Royal Society, and the University of Warwick. M.M. was supported by funds from the Wellcome Trust India Alliance and the Tata Institute of Fundamental Research, Mumbai, India.
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Huang, J., Mishra, M., Palani, S., Chew, T.G., Balasubramanian, M.K. (2016). Isolation of Cytokinetic Actomyosin Rings from Saccharomyces cerevisiae and Schizosaccharomyces pombe . In: Sanchez-Diaz, A., Perez, P. (eds) Yeast Cytokinesis. Methods in Molecular Biology, vol 1369. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3145-3_10
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DOI: https://doi.org/10.1007/978-1-4939-3145-3_10
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3144-6
Online ISBN: 978-1-4939-3145-3
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