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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.

SUBMITTER: Buguliskis JS

PROVIDER: S-EPMC2858701 | biostudies-literature | 2010 Apr

REPOSITORIES: biostudies-literature

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Rhomboid 4 (ROM4) affects the processing of surface adhesins and facilitates host cell invasion by Toxoplasma gondii.

Buguliskis Jeffrey S JS Brossier Fabien F Shuman Joel J Sibley L David LD

PLoS pathogens 20100422 4

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 condi ...[more]

PMID: 20421941

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Project description:Host cell invasion by Toxoplasma gondii and other apicomplexan parasites requires transmembrane adhesins that mediate binding to receptors on the substrate and host cell to facilitate motility and invasion. Rhomboid proteases (ROMs) are thought to cleave adhesins within their transmembrane segments, thus allowing the parasite to disengage from receptors and completely enter the host cell. To examine the specific roles of individual ROMs during invasion, we generated single, double, and triple knockouts for the three ROMs expressed in T. gondii tachyzoites. Analysis of these mutants demonstrated that ROM4 is the primary protease involved in adhesin processing and host cell invasion, whereas ROM1 or ROM5 plays negligible roles in these processes. Deletion of ROM4 blocked the shedding of adhesins such as MIC2 (microneme protein 2), causing them to accumulate on the surface of extracellular parasites. Increased surface adhesins led to nonproductive attachment, altered gliding motility, impaired moving junction formation, and reduced invasion efficiency. Despite the importance of ROM4 for efficient invasion, mutants lacking all three ROMs were viable and MIC2 was still efficiently removed from the surface of invaded mutant parasites, implying the existence of ROM-independent mechanisms for adhesin removal during invasion. Collectively, these results suggest that although ROM processing of adhesins is not absolutely essential, it is important for efficient host cell invasion by T. gondii. Importance: Apicomplexan parasites such as Toxoplasma gondii express surface proteins that bind host cell receptors to aid invasion. Many of these adhesins are subject to cleavage by rhomboid proteases (ROMs) within their transmembrane segments during invasion. Previous studies have demonstrated the importance of adhesin cleavage for parasite invasion and proposed that the ROMs responsible for processing would be essential for parasite survival. In T. gondii, ROM5 was thought to be the critical ROM for adhesin shedding due to its robust protease activity in vitro and posterior localization on the parasite surface. Here, we knocked out all three ROMs in T. gondii tachyzoites and found that ROM4, but not ROM5, was key for adhesin cleavage. However, none of the ROMs individually or in combination was essential for cell entry, further emphasizing that essential pathways such as invasion typically rely on redundant pathways to ensure survival.

Dataset Information

Rhomboid 4 (ROM4) affects the processing of surface adhesins and facilitates host cell invasion by Toxoplasma gondii.

Publications

Rhomboid 4 (ROM4) affects the processing of surface adhesins and facilitates host cell invasion by Toxoplasma gondii.

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