title: Actin depolymerizing factor controls actin turnover and gliding motility in Toxoplasma gondii.
abstract: Apicomplexan parasites rely on actin-based gliding motility to move across the substratum, cross biological barriers, and invade their host cells. Gliding motility depends on polymerization of parasite actin filaments, yet ∼98% of actin is nonfilamentous in resting parasites. Previous studies suggest that the lack of actin filaments in the parasite is due to inherent instability, leaving uncertain the role of actin-binding proteins in controlling dynamics. We have previously shown that the single allele of Toxoplasma gondii actin depolymerizing factor (TgADF) has strong actin monomer-sequestering and weak filament-severing activities in vitro. Here we used a conditional knockout strategy to investigate the role of TgADF in vivo. Suppression of TgADF led to accumulation of actin-rich filaments that were detected by immunofluorescence and electron microscopy. Parasites deficient in TgADF showed reduced speed of motility, increased aberrant patterns of motion, and inhibition of sustained helical gliding. Lack of TgADF also led to severe defects in entry and egress from host cells, thus blocking infection in vitro. These studies establish that the absence of stable actin structures in the parasite are not simply the result of intrinsic instability, but that TgADF is required for the rapid turnover of parasite actin filaments, gliding motility, and cell invasion.
authors.affiliation: Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Simren
authors.initials: S
authors.lastname: Mehta
authors.forename: L David
authors.initials: LD
authors.lastname: Sibley
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism*; D000199:Actins / Q000378:metabolism; D018929:Cell Culture Techniques; D002465:Cell Movement; D003599:Cytoskeleton / Q000378:metabolism; D051345:Destrin / Q000235:genetics / Q000378:metabolism*; D005347:Fibroblasts / Q000469:parasitology; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D055786:Gene Knockout Techniques; D006790:Host-Parasite Interactions; D006801:Humans; D008297:Male; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism*; D008854:Microscopy, Electron; D058105:Polymerization; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D014122:Toxoplasma / Q000502:physiology
chemical_list: D000199:Actins; D051345:Destrin; D008840:Microfilament Proteins; D015800:Protozoan Proteins