Tropomyosin and myosin-II cellular levels promote actomyosin ring assembly in fission yeast.
Myosin-II (Myo2p) and tropomyosin are essential for contractile ring formation and cytokinesis in fission yeast. Here we used a combination of in vivo and in vitro approaches to understand how these proteins function at contractile rings. We find that ring assembly is delayed in Myo2p motor and tropomyosin mutants, but occurs prematurely in cells engineered to express two copies of myo2. Thus, the timing of ring assembly responds to changes in Myo2p cellular levels and motor activity, and the emergence of tropomyosin-bound actin filaments. Doubling Myo2p levels suppresses defects in ring assembly associated with a tropomyosin mutant, suggesting a role for tropomyosin in maximizing Myo2p function. Correspondingly, tropomyosin increases Myo2p actin affinity and ATPase activity and promotes Myo2p-driven actin filament gliding in motility assays. Tropomyosin achieves this by favoring the strong actin-bound state of Myo2p. This mode of regulation reflects a role for tropomyosin in specifying and stabilizing actomyosin interactions, which facilitates contractile ring assembly in the fission yeast system.
Stark
Benjamin C
BC
Department of Molecular Physiology and Biophysics, University of Vermont, Burlington, VT 05405, USA.
Sladewski
Thomas E
TE
Pollard
Luther W
LW
Lord
Matthew
M
D000205:Actomyosin / Q000235:genetics / Q000378:metabolism*; D048749:Cytokinesis / Q000502:physiology*; D036681:Fluorescence Recovery After Photobleaching; D018995:Myosin Heavy Chains / Q000235:genetics / Q000378:metabolism*; D024462:Myosin Type II / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012568:Schizosaccharomyces / Q000166:cytology* / Q000502:physiology; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000378:metabolism*; D014335:Tropomyosin / Q000235:genetics / Q000378:metabolism*
C434211:MYO2 protein, S pombe; D011993:Recombinant Fusion Proteins; D029702:Schizosaccharomyces pombe Proteins; D014335:Tropomyosin; D000205:Actomyosin; D024462:Myosin Type II; D018995:Myosin Heavy Chains