Abstract

Apicomplexan parasites, including Toxoplasma gondii and Plasmodium sp., are obligate intracellular protozoa. They enter into a host cell by attaching to and then creating an invagination in the host cell plasma membrane. Contact between parasite and host plasma membranes occurs in the form of a ring-shaped moving junction that begins at the anterior end of the parasite and then migrates posteriorly. The resulting invagination of host plasma membrane creates a parasitophorous vacuole that completely envelops the now intracellular parasite. At the start of this process, apical membrane antigen 1 (AMA1) is released onto the parasite surface from specialized secretory organelles called micronemes. The T. gondii version of this protein, TgAMA1, has been shown to be essential for invasion but its exact role has not previously been determined. We identify here a trio of proteins that associate with TgAMA1, at least one of which associates with TgAMA1 at the moving junction. Surprisingly, these new proteins derive not from micronemes, but from the anterior secretory organelles known as rhoptries and specifically, for at least two, from the neck portion of these club-shaped structures. Homologues for these AMA1-associated proteins are found throughout the Apicomplexa strongly suggesting that this moving junction apparatus is a conserved feature of this important class of parasites. Differences between the contributing proteins in different species may, in part, be the result of selective pressure from the different niches occupied by these parasites.

Highlights

  • Toxoplasma gondii is an obligate intracellular parasite that belongs to the phylum Apicomplexa

  • The specificity of the T. gondii version of AMA1 (TgAMA1)-associating protein (AAP) association was further demonstrated by immunoblotting the material that co-precipitated with anti-TgAMA1 and probing for the highly abundant MIC2, GRA7, ROP1, or ROP2/3/4 proteins

  • We describe the coordinated secretion of rhoptry necks and micronemes to form a complex of apparently four proteins, TgAMA1, RON4, RON2 and Ts4705, at least of two of which interact at the moving junction (MJ) of invading parasites

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Summary

Introduction

Toxoplasma gondii is an obligate intracellular parasite that belongs to the phylum Apicomplexa. All members of this phylum exhibit a similar process for invading into host cells [1]. The parasites establish an intimate association, involving reorientation to put the apical secretory structures in contact with the host membrane [2]. This reorientation is coincident with a transient spike in host plasma membrane conductance, consistent with a break in the bilayer [3]. A moving junction (MJ), where the host and parasite plasma membranes are in intimate contact, is formed

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