title: High-resolution temporal analysis reveals a functional timeline for the molecular regulation of cytokinesis.
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.
authors: Department of Pathology and Cell Biology, Columbia University, New York, NY 10032, USA.
authors: Tim
authors: T
authors: Davies
authors: Department of Pathology and Cell Biology, Columbia University, New York, NY 10032, USA.
authors: Shawn N
authors: SN
authors: Jordan
authors: Department of Pathology and Cell Biology, Columbia University, New York, NY 10032, USA.
authors: Vandana
authors: V
authors: Chand
authors: Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, IL 60637, USA.
authors: Jennifer A
authors: JA
authors: Sees
authors: CNRS, Institut Jacques Monod, Univ. P7, 75205 Paris CEDEX 13, France.
authors: Kimberley
authors: K
authors: Laband
authors: Molecular and Cellular Biology Unit, Instituto de Biologia Molecular e Celular (IBMC), 4150-180 Porto, Portugal.
authors: Ana X
authors: AX
authors: Carvalho
authors: Department of Genetics and Development, Columbia University, New York, NY 10032, USA.
authors: Mimi
authors: M
authors: Shirasu-Hiza
authors: 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.
authors: David R
authors: DR
authors: Kovar
authors: CNRS, Institut Jacques Monod, Univ. P7, 75205 Paris CEDEX 13, France.
authors: Julien
authors: J
authors: Dumont
authors: Department of Pathology and Cell Biology, Columbia University, New York, NY 10032, USA. Electronic address: jcc2210@columbia.edu.
authors: Julie C
authors: JC
authors: Canman
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D064107:Aurora Kinase B / Q000378:metabolism; D017173:Caenorhabditis elegans / Q000166:cytology / Q000378:metabolism* / Q000502:physiology; D029742:Caenorhabditis elegans Proteins / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D048749:Cytokinesis; D005326:Fetal Proteins / Q000378:metabolism; D000080865:Formins; D006361:Heating / Q000379:methods; D006358:Hot Temperature; D008840:Microfilament Proteins / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D024462:Myosin Type II / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D061848:Optical Imaging / Q000379:methods; D055550:Protein Stability
chemical_list: D000199:Actins; D029742:Caenorhabditis elegans Proteins; D002868:Chromosomal Proteins, Non-Histone; D005326:Fetal Proteins; D000080865:Formins; C490517:MKLP1 protein, C elegans; D008840:Microfilament Proteins; D008869:Microtubule-Associated Proteins; D009687:Nuclear Proteins; D064107:Aurora Kinase B; D024462:Myosin Type II