Actin filament polymerization regulates gliding motility by apicomplexan parasites.
Host cell entry by Toxoplasma gondii depends critically on actin filaments in the parasite, yet paradoxically, its actin is almost exclusively monomeric. In contrast to the absence of stable filaments in conventional samples, rapid-freeze electron microscopy revealed that actin filaments were formed beneath the plasma membrane of gliding parasites. To investigate the role of actin filaments in motility, we treated parasites with the filament-stabilizing drug jasplakinolide (JAS) and monitored the distribution of actin in live and fixed cells using yellow fluorescent protein (YFP)-actin. JAS treatment caused YFP-actin to redistribute to the apical and posterior ends, where filaments formed a spiral pattern subtending the plasma membrane. Although previous studies have suggested that JAS induces rigor, videomicroscopy demonstrated that JAS treatment increased the rate of parasite gliding by approximately threefold, indicating that filaments are rate limiting for motility. However, JAS also frequently reversed the normal direction of motility, disrupting forward migration and cell entry. Consistent with this alteration, subcortical filaments in JAS-treated parasites occurred in tangled plaques as opposed to the straight, roughly parallel orientation observed in control cells. These studies reveal that precisely controlled polymerization of actin filaments imparts the correct timing, duration, and directionality of gliding motility in the Apicomplexa.
Wetzel
D M
DM
Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Håkansson
S
S
Hu
K
K
Roos
D
D
Sibley
L D
LD
D008841:Actin Cytoskeleton / Q000378:metabolism*; D000199:Actins / Q000378:metabolism*; D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism; D015139:Blotting, Southern; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D020285:Cryoelectron Microscopy; D003599:Cytoskeleton / Q000648:ultrastructure; D003600:Cytosol / Q000378:metabolism; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D008840:Microfilament Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D018715:Microscopy, Video; D009068:Movement; D010957:Plasmids / Q000378:metabolism; D011817:Rabbits; D013997:Time Factors; D014122:Toxoplasma / Q000378:metabolism / Q000472:pathogenicity*
D000199:Actins; D001426:Bacterial Proteins; D008164:Luminescent Proteins; D008840:Microfilament Proteins; C054562:yellow fluorescent protein, Bacteria