Tim Davies, Shawn N Jordan, Vandana Chand, Jennifer A Sees, Kimberley Laband, Ana X Carvalho, Mimi Shirasu-Hiza, David R Kovar, Julien Dumont, Julie C Canman
Developmental cell 2014 Jul 28To 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. Copyright © 2014 Elsevier Inc. All rights reserved.
Tim Davies, Shawn N Jordan, Vandana Chand, Jennifer A Sees, Kimberley Laband, Ana X Carvalho, Mimi Shirasu-Hiza, David R Kovar, Julien Dumont, Julie C Canman. High-resolution temporal analysis reveals a functional timeline for the molecular regulation of cytokinesis. Developmental cell. 2014 Jul 28;30(2):209-23
PMID: 25073157
View Full Text