Differential localization of Acanthamoeba myosin I isoforms.
Acanthamoeba myosins IA and IB were localized by immunofluorescence and immunoelectron microscopy in vegetative and phagocytosing cells and the total cell contents of myosins IA, IB, and IC were quantified by immunoprecipitation. The quantitative distributions of the three myosin I isoforms were then calculated from these data and the previously determined localization of myosin IC. Myosin IA occurs almost exclusively in the cytoplasm, where it accounts for approximately 50% of the total myosin I, in the cortex beneath phagocytic cups and in association with small cytoplasmic vesicles. Myosin IB is the predominant isoform associated with the plasma membrane, large vacuole membranes and phagocytic membranes and accounts for almost half of the total myosin I in the cytoplasm. Myosin IC accounts for a significant fraction of the total myosin I associated with the plasma membrane and large vacuole membranes and is the only myosin I isoform associated with the contractile vacuole membrane. These data suggest that myosin IA may function in cytoplasmic vesicle transport and myosin I-mediated cortical contraction, myosin IB in pseudopod extension and phagocytosis, and myosin IC in contractile vacuole function. In addition, endogenous and exogenously added myosins IA and IB appeared to be associated with the cytoplasmic surface of different subpopulations of purified plasma membranes implying that the different myosin I isoforms are targeted to specific membrane domains through a mechanism that involves more than the affinity of the myosins for anionic phospholipids.
Baines
I C
IC
Laboratory of Cell Biology, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892.
Brzeska
H
H
Korn
E D
ED
D000048:Acanthamoeba / Q000737:chemistry* / Q000648:ultrastructure*; D000818:Animals; D001692:Biological Transport; D002451:Cell Compartmentation; D002462:Cell Membrane / Q000737:chemistry; D005455:Fluorescent Antibody Technique; D014644:Genetic Variation; D007150:Immunohistochemistry; D007425:Intracellular Membranes / Q000737:chemistry; D008565:Membrane Proteins / Q000302:isolation & purification; D016253:Microscopy, Immunoelectron; D009218:Myosins / Q000276:immunology / Q000302:isolation & purification*; D010587:Phagocytosis; D014617:Vacuoles / Q000737:chemistry
D008565:Membrane Proteins; D009218:Myosins