The Conoid Associated Motor MyoH Is Indispensable for Toxoplasma gondii Entry and Exit from Host Cells

PLoS Pathog. 2016 Jan 13;12(1):e1005388. doi: 10.1371/journal.ppat.1005388. eCollection 2016 Jan.

Abstract

Many members of the phylum of Apicomplexa have adopted an obligate intracellular life style and critically depend on active invasion and egress from the infected cells to complete their lytic cycle. Toxoplasma gondii belongs to the coccidian subgroup of the Apicomplexa, and as such, the invasive tachyzoite contains an organelle termed the conoid at its extreme apex. This motile organelle consists of a unique polymer of tubulin fibres and protrudes in both gliding and invading parasites. The class XIV myosin A, which is conserved across the Apicomplexa phylum, is known to critically contribute to motility, invasion and egress from infected cells. The MyoA-glideosome is anchored to the inner membrane complex (IMC) and is assumed to translocate the components of the circular junction secreted by the micronemes and rhoptries, to the rear of the parasite. Here we comprehensively characterise the class XIV myosin H (MyoH) and its associated light chains. We show that the 3 alpha-tubulin suppressor domains, located in MyoH tail, are necessary to anchor this motor to the conoid. Despite the presence of an intact MyoA-glideosome, conditional disruption of TgMyoH severely compromises parasite motility, invasion and egress from infected cells. We demonstrate that MyoH is necessary for the translocation of the circular junction from the tip of the parasite, where secretory organelles exocytosis occurs, to the apical position where the IMC starts. This study attributes for the first time a direct function of the conoid in motility and invasion, and establishes the indispensable role of MyoH in initiating the first step of motility along this unique organelle, which is subsequently relayed by MyoA to enact effective gliding and invasion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western
  • Chlorocebus aethiops
  • Fluorescent Antibody Technique, Indirect
  • Gene Knockout Techniques
  • Host-Parasite Interactions / physiology*
  • Humans
  • Immunoprecipitation
  • Microscopy, Confocal
  • Microscopy, Electron, Transmission
  • Myosin Heavy Chains / metabolism*
  • Organelles
  • Protozoan Proteins / metabolism*
  • Toxoplasma / cytology
  • Toxoplasma / pathogenicity*
  • Toxoplasmosis / metabolism*
  • Transfection
  • Vero Cells

Substances

  • Protozoan Proteins
  • Myosin Heavy Chains

Grants and funding

This study was supported by the Swiss National Foundation (recipient: DSF, http://p3.snf.ch/project-147118). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.