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Dev Cell. 2014 Jul 28;30(2):209-23. doi: 10.1016/j.devcel.2014.05.009.

High-resolution temporal analysis reveals a functional timeline for the molecular regulation of cytokinesis.

Author information

1
Department of Pathology and Cell Biology, Columbia University, New York, NY 10032, USA.
2
Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, IL 60637, USA.
3
CNRS, Institut Jacques Monod, Univ. P7, 75205 Paris CEDEX 13, France.
4
Molecular and Cellular Biology Unit, Instituto de Biologia Molecular e Celular (IBMC), 4150-180 Porto, Portugal.
5
Department of Genetics and Development, Columbia University, New York, NY 10032, USA.
6
Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, IL 60637, USA; Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL 60637, USA.
7
Department of Pathology and Cell Biology, Columbia University, New York, NY 10032, USA. Electronic address: jcc2210@columbia.edu.

Abstract

To take full advantage of fast-acting temperature-sensitive mutations, thermal control must be extremely rapid. We developed the Therminator, a device capable of shifting sample temperature in ~17 s while simultaneously imaging cell division in vivo. Applying this technology to six key regulators of cytokinesis, we found that each has a distinct temporal requirement in the Caenorhabditis elegans zygote. Specifically, myosin-II is required throughout cytokinesis until contractile ring closure. In contrast, formin-mediated actin nucleation is only required during assembly and early contractile ring constriction. Centralspindlin is required to maintain division after ring closure, although its GAP activity is only required until just prior to closure. Finally, the chromosomal passenger complex is required for cytokinesis only early in mitosis, but not during metaphase or cytokinesis. Together, our results provide a precise functional timeline for molecular regulators of cytokinesis using the Therminator, a powerful tool for ultra-rapid protein inactivation.

PMID:
25073157
PMCID:
PMC4245203
DOI:
10.1016/j.devcel.2014.05.009
[Indexed for MEDLINE]
Free PMC Article

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