The role of myosin II motor activity in distributing myosin asymmetrically and coupling protrusive activity to cell translocation.
Nonmuscle myosin IIA and IIB distribute preferentially toward opposite ends of migrating endothelial cells. To understand the mechanism and function of this behavior, myosin II was examined in cells treated with the motor inhibitor, blebbistatin. Blebbistatin at > or = 30 microM inhibited anterior redistribution of myosin IIA, with 100 microM blebbistatin causing posterior accumulation. Posterior accumulation of myosin IIB was unaffected. Time-lapse cinemicrography showed myosin IIA entering lamellipodia shortly after their formation, but failing to move into lamellipodia in blebbistatin. Thus, myosin II requires motor activity to move forward onto F-actin in protrusions. However, this movement is inhibited by myosin filament assembly, because whole myosin was delayed relative to a tailless fragment. Inhibiting myosin's forward movement reduced coupling between protrusive activity and translocation of the cell body: In untreated cells, body movement followed advancing lamellipodia, whereas blebbistatin-treated cells extended protrusions without displacement of the body or with a longer delay before movement. Anterior cytoplasm of blebbistatin-treated cells contained disorganized bundles of parallel microfilaments, but anterior F-actin bundles in untreated cells were mostly oriented perpendicular to movement. Myosin II may ordinarily move anteriorly on actin filaments and pull crossed filaments into antiparallel bundles, with the resulting realignment pulling the cell body forward.
Kolega
John
J
Department of Pathology and Anatomical Sciences, Division of Anatomy and Cell Biology, State University of New York at Buffalo School of Medicine and Biomedical Sciences, Buffalo, NY 14214, USA. kolega@buffalo.edu
D008841:Actin Cytoskeleton / Q000187:drug effects / Q000648:ultrastructure; D000199:Actins / Q000378:metabolism; D000818:Animals; D002417:Cattle; D002465:Cell Movement; D016764:Cell Polarity; D002478:Cells, Cultured; D003593:Cytoplasm / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000502:physiology / Q000648:ultrastructure; D006576:Heterocyclic Compounds, 4 or More Rings / Q000494:pharmacology*; D008954:Models, Biological; D024462:Myosin Type II / Q000378:metabolism* / Q000502:physiology*; D020033:Protein Isoforms
D000199:Actins; D006576:Heterocyclic Compounds, 4 or More Rings; D020033:Protein Isoforms; C472645:blebbistatin; D024462:Myosin Type II