title: Convergent use of RhoGAP toxins by eukaryotic parasites and bacterial pathogens.
abstract: Inactivation of host Rho GTPases is a widespread strategy employed by bacterial pathogens to manipulate mammalian cellular functions and avoid immune defenses. Some bacterial toxins mimic eukaryotic Rho GTPase-activating proteins (GAPs) to inactivate mammalian GTPases, probably as a result of evolutionary convergence. An intriguing question remains whether eukaryotic pathogens or parasites may use endogenous GAPs as immune-suppressive toxins to target the same key genes as bacterial pathogens. Interestingly, a RhoGAP domain-containing protein, LbGAP, was recently characterized from the parasitoid wasp Leptopilina boulardi, and shown to protect parasitoid eggs from the immune response of Drosophila host larvae. We demonstrate here that LbGAP has structural characteristics of eukaryotic RhoGAPs but that it acts similarly to bacterial RhoGAP toxins in mammals. First, we show by immunocytochemistry that LbGAP enters Drosophila immune cells, plasmatocytes and lamellocytes, and that morphological changes in lamellocytes are correlated with the quantity of LbGAP they contain. Demonstration that LbGAP displays a GAP activity and specifically interacts with the active, GTP-bound form of the two Drosophila Rho GTPases Rac1 and Rac2, both required for successful encapsulation of Leptopilina eggs, was then achieved using biochemical tests, yeast two-hybrid analysis, and GST pull-down assays. In addition, we show that the overall structure of LbGAP is similar to that of eukaryotic RhoGAP domains, and we identify distinct residues involved in its interaction with Rac GTPases. Altogether, these results show that eukaryotic parasites can use endogenous RhoGAPs as virulence factors and that despite their differences in sequence and structure, eukaryotic and bacterial RhoGAP toxins are similarly used to target the same immune pathways in insects and mammals.
authors.affiliation: UMR 1112 UNSA-INRA Résponses des Organismes aux Stress Environnementaux, Sophia-Antipolis, France.
authors.forename: Dominique
authors.initials: D
authors.lastname: Colinet
authors.forename: Antonin
authors.initials: A
authors.lastname: Schmitz
authors.forename: Delphine
authors.initials: D
authors.lastname: Depoix
authors.forename: Didier
authors.initials: D
authors.lastname: Crochard
authors.forename: Marylène
authors.initials: M
authors.lastname: Poirié
mesh_terms: D000818:Animals; D001419:Bacteria / Q000235:genetics; D029721:Drosophila Proteins / Q000378:metabolism; D004331:Drosophila melanogaster / Q000276:immunology / Q000382:microbiology / Q000469:parasitology*; D019143:Evolution, Molecular; D020690:GTPase-Activating Proteins / Q000235:genetics / Q000378:metabolism*; D006434:Hemocytes / Q000382:microbiology / Q000469:parasitology; D054884:Host-Pathogen Interactions; D007814:Larva / Q000235:genetics / Q000254:growth & development / Q000276:immunology; D008969:Molecular Sequence Data; D016296:Mutagenesis; D020798:Two-Hybrid System Techniques; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism*; D014863:Wasps / Q000235:genetics / Q000254:growth & development* / Q000276:immunology; D020744:rac GTP-Binding Proteins / Q000378:metabolism; D000097766:RAC2 GTP-Binding Protein
chemical_list: D029721:Drosophila Proteins; D020690:GTPase-Activating Proteins; D037521:Virulence Factors; D020744:rac GTP-Binding Proteins; D000097766:RAC2 GTP-Binding Protein; C090146:Rac1 protein, Drosophila; C085379:rho GTPase-activating protein