Myosin heavy chain phosphorylation sites regulate myosin localization during cytokinesis in live cells.
Conventional myosin II plays a fundamental role in the process of cytokinesis where, in the form of bipolar thick filaments, it is thought to be the molecular motor that generates the force necessary to divide the cell. In Dictyostelium, the formation of thick filaments is regulated by the phosphorylation of three threonine residues in the tail region of the myosin heavy chain. We report here on the effects of this regulation on the localization of myosin in live cells undergoing cytokinesis. We imaged fusion proteins of the green-fluorescent protein with wild-type myosin and with myosins where the three critical threonines had been changed to either alanine or aspartic acid. We provide evidence that thick filament formation is required for the accumulation of myosin in the cleavage furrow and that if thick filaments are overproduced, this accumulation is markedly enhanced. This suggests that myosin localization in dividing cells is regulated by myosin heavy chain phosphorylation.
Sabry
J H
JH
Department of Biochemistry, Stanford University School of Medicine, Stanford, CA 94305, USA.
Moores
S L
SL
Ryan
S
S
Zang
J H
JH
Spudich
J A
JA
D000199:Actins / Q000378:metabolism; D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D001692:Biological Transport; D002455:Cell Division; D002465:Cell Movement; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival; D002478:Cells, Cultured; D004023:Dictyostelium / Q000166:cytology* / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D049452:Green Fluorescent Proteins; D007700:Kinetics; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018995:Myosin Heavy Chains / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009218:Myosins / Q000378:metabolism*; D010766:Phosphorylation; D010769:Phosphothreonine / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013912:Threonine / Q000235:genetics / Q000378:metabolism
D000199:Actins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D010769:Phosphothreonine; D049452:Green Fluorescent Proteins; D013912:Threonine; D018995:Myosin Heavy Chains; D009218:Myosins