title: Measurements of Myosin-II Motor Activity During Cytokinesis in Fission Yeast.
abstract: Fission yeast myosin-II (Myo2p) represents the critical actin-based motor protein that drives actomyosin ring assembly and constriction during cytokinesis. We detail three different methods to measure Myo2p motor function. Actin-activated ATPases provide a readout of actomyosin ATPase motor activity in a bulk assay; actin filament motility assays reveal the speed and efficiency of myosin-driven actin filament gliding (when motors are anchored); myosin-bead motility assays reveal the speed and efficiency of myosin ensembles traveling along actin filaments (when actin is anchored). Collectively, these methods allow us to combine the standard in vivo approaches common to fission yeast with in vitro biochemical methods to learn more about the mechanistic action of myosin-II during cytokinesis.
authors.affiliation: Department of Molecular Physiology & Biophysics, University of Vermont, Burlington, VT, 05405, USA.
authors.forename: Qing
authors.initials: Q
authors.lastname: Tang
authors.affiliation: Department of Molecular Physiology & Biophysics, University of Vermont, Burlington, VT, 05405, USA.
authors.forename: Luther W
authors.initials: LW
authors.lastname: Pollard
authors.affiliation: Department of Molecular Physiology & Biophysics, University of Vermont, Burlington, VT, 05405, USA. matthew.lord@uvm.edu.
authors.forename: Matthew
authors.initials: M
authors.lastname: Lord
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D048749:Cytokinesis / Q000235:genetics; D015870:Gene Expression; D008856:Microscopy, Fluorescence; D057054:Molecular Imaging / Q000379:methods; D024462:Myosin Type II / Q000235:genetics / Q000378:metabolism*; D012568:Schizosaccharomyces / Q000502:physiology*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D029702:Schizosaccharomyces pombe Proteins; D024462:Myosin Type II
keywords: Actin -activated ATPase assay; Actin filament motility assay; Colorimetric assay; Epifluorescence microscopy; Myosin-bead motility assay s; Protein-fluorophore s; Total internal reflection fluorescence microscopy