{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,19]],"date-time":"2026-09-19T05:15:16Z","timestamp":1789794916000,"version":"4.0.1"},"reference-count":47,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,7,9]],"date-time":"2025-07-09T00:00:00Z","timestamp":1752019200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["STE 2348\/2-1 , STE 2348\/2-2"],"award-info":[{"award-number":["STE 2348\/2-1 , STE 2348\/2-2"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:p>\n                    <jats:italic>Toxoplasma gondii<\/jats:italic>\n                    (\n                    <jats:italic>T. gondii<\/jats:italic>\n                    ) virulence in mice depends on different multiprotein complexes that assemble at the parasitophorous vacuole membrane (PVM) of the parasite. Individual rhoptry proteins within these complexes inhibit different Immunity-Related GTPases (IRG proteins). The rhoptry pseudokinase ROP5 is a central element to achieve IRG-specific rhoptry kinase activity and\/or efficient complex formation. The\n                    <jats:italic>rop5<\/jats:italic>\n                    locus of each of the canonical\n                    <jats:italic>T. gondii<\/jats:italic>\n                    strains encodes three major isoforms, ROP5A, ROP5B and ROP5C, and was shown to have the largest impact on virulence. By reverse genetics, we have generated\n                    <jats:italic>T. gondii<\/jats:italic>\n                    strains expressing either ROP5A, ROP5B or ROP5C in a RH\u0394\n                    <jats:italic>rop5<\/jats:italic>\n                    genetic background and demonstrate that ROP5B is mainly responsible for heightened virulence of type I\n                    <jats:italic>T. gondii<\/jats:italic>\n                    in laboratory strains of mice.\n                    <jats:italic>In vivo<\/jats:italic>\n                    virulence correlates with diminished vacuolar IRG protein loading and parasite control\n                    <jats:italic>in vitro<\/jats:italic>\n                    only in presence of ROP5B but not ROP5A or ROP5C. Our results suggest that ROP5A and ROP5C isoforms might have co-evolved with IRG proteins or other host cell resistance factors in evolutionarily important intermediate hosts beyond\n                    <jats:italic>Mus musculus<\/jats:italic>\n                    . The same parasite effectors that inhibit IRG protein accumulation and function reduce the vacuolar amount of Guanylate Binding Proteins (GBP proteins). However, a parasite effector targeting a GBP protein at the PVM has not been described yet. Using two different approaches, Yeast Two-Hybrid analysis and Protein-fragment complementation assay, we here identified three heterologous IRG:GBP pairs, GBP6:Irgb10, GBP5:Irgb10, GBP5:Irgb6, and demonstrate that the accumulation of these GTPases at the PVM is interdependent. Our results offer a novel perspective on the IRG and GBP protein-mediated control of\n                    <jats:italic>T. gondii<\/jats:italic>\n                    infections and may further advance the investigation of GBP-specific\n                    <jats:italic>T. gondii<\/jats:italic>\n                    effectors.\n                  <\/jats:p>","DOI":"10.3389\/fimmu.2025.1593785","type":"journal-article","created":{"date-parts":[[2025,7,10]],"date-time":"2025-07-10T05:46:09Z","timestamp":1752126369000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":6,"title":["Toxoplasma gondii virulence in mice is determined by the pseudokinase ROP5B and countered by an IRG-GBP protein 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