title: Dense granule trafficking in Toxoplasma gondii requires a unique class 27 myosin and actin filaments.
abstract: The survival of Toxoplasma gondii within its host cell requires protein release from secretory vesicles, called dense granules, to maintain the parasite's intracellular replicative niche. Despite the importance of DGs, nothing is known about the mechanisms underlying their transport. In higher eukaryotes, secretory vesicles are transported to the plasma membrane by molecular motors moving on their respective cytoskeletal tracks (i.e., microtubules and actin). Because the organization of these cytoskeletal structures differs substantially in T. gondii, the molecular motor dependence of DG trafficking is far from certain. By imaging the motions of green fluorescent protein-tagged DGs in intracellular parasites with high temporal and spatial resolution, we show through a combination of molecular genetics and chemical perturbations that directed DG transport is independent of microtubules and presumably their kinesin/dynein motors. However, directed DG transport is dependent on filamentous actin and a unique class 27 myosin, TgMyoF, which has structural similarity to myosin V, the prototypical cargo transporter. Actomyosin DG transport was unexpected, since filamentous parasite actin has yet to be visualized in vivo due in part to the prevailing model that parasite actin forms short, unstable filaments. Thus our data uncover new critical roles for these essential proteins in the lytic cycle of this devastating pathogen.
authors.affiliation: Department of Molecular Physiology and Biophysics, University of Vermont, Burlington, VT 05405 aheaslip@uvm.edu david.warshaw@uvm.edu.
authors.forename: Aoife T
authors.initials: AT
authors.lastname: Heaslip
authors.affiliation: Department of Molecular Physiology and Biophysics, University of Vermont, Burlington, VT 05405.
authors.forename: Shane R
authors.initials: SR
authors.lastname: Nelson
authors.affiliation: Department of Molecular Physiology and Biophysics, University of Vermont, Burlington, VT 05405 aheaslip@uvm.edu david.warshaw@uvm.edu.
authors.forename: David M
authors.initials: DM
authors.lastname: Warshaw
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002465:Cell Movement; D022162:Cytoplasmic Vesicles / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D016547:Kinesins / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism; D024701:Myosin Type V / Q000378:metabolism*; D009218:Myosins; D021381:Protein Transport; D022142:Secretory Vesicles / Q000378:metabolism / Q000502:physiology; D062211:Spatio-Temporal Analysis; D014122:Toxoplasma / Q000235:genetics / Q000378:metabolism*
chemical_list: D000199:Actins; D024701:Myosin Type V; D009218:Myosins; D016547:Kinesins