title: Rhomboid 4 (ROM4) affects the processing of surface adhesins and facilitates host cell invasion by Toxoplasma gondii.
abstract: Host cell attachment by Toxoplasma gondii is dependent on polarized secretion of apical adhesins released from the micronemes. Subsequent translocation of these adhesive complexes by an actin-myosin motor powers motility and host cell invasion. Invasion and motility are also accompanied by shedding of surface adhesins by intramembrane proteolysis. Several previous studies have implicated rhomboid proteases in this step; however, their precise roles in vivo have not been elucidated. Using a conditional knockout strategy, we demonstrate that TgROM4 participates in processing of surface adhesins including MIC2, AMA1, and MIC3. Suppression of TgROM4 led to decreased release of the adhesin MIC2 into the supernatant and concomitantly increased the surface expression of this and a subset of other adhesins. Suppression of TgROM4 resulted in disruption of normal gliding, with the majority of parasites twirling on their posterior ends. Parasites lacking TgROM4 bound better to host cells, but lost the ability to apically orient and consequently most failed to generate a moving junction; hence, invasion was severely impaired. Our findings indicate that TgROM4 is involved in shedding of micronemal proteins from the cell surface. Down regulation of TgROM4 disrupts the normal apical-posterior gradient of adhesins that is important for efficient cell motility and invasion of host cells by T. gondii.
authors.affiliation: Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri, United States of America.
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Buguliskis
authors.forename: Fabien
authors.initials: F
authors.lastname: Brossier
authors.forename: Joel
authors.initials: J
authors.lastname: Shuman
authors.forename: L David
authors.initials: LD
authors.lastname: Sibley
mesh_terms: D000953:Antigens, Protozoan / Q000378:metabolism; D002469:Cell Separation; D002478:Cells, Cultured; D005347:Fibroblasts / Q000469:parasitology; D005434:Flow Cytometry; D055786:Gene Knockout Techniques; D006790:Host-Parasite Interactions / Q000502:physiology*; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014122:Toxoplasma / Q000378:metabolism / Q000472:pathogenicity*
chemical_list: C506564:AMA1 protein, Toxoplasma gondii; D000953:Antigens, Protozoan; C408878:MIC2 protein, Toxoplasma gondii; C421382:MIC3 protein, Toxoplasma gondii; D008565:Membrane Proteins; D015800:Protozoan Proteins