{"gene":"TG_237015","gene_symbol":"GRA43","run_date":"2026-09-08T15:31:32+00:00","timeline":{"discoveries":[{"year":2019,"finding":"GRA43 is a parasitophorous vacuole-localized dense granule protein individually required for induction of Lewis rat macrophage pyroptosis; deletion of GRA43 greatly reduces macrophage cell death and enhances parasite replication. Epistasis analysis (single, double, and triple deletion mutants showing similar viability levels) places GRA35, GRA42, and GRA43 in the same pathway.","method":"Chemical mutagenesis screen, whole-genome sequencing, CRISPR/knockout (Δgra43), cell viability assays, in vivo infection of Lewis rats","journal":"mBio","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean KO with defined cellular phenotype, genetic epistasis via single/double/triple deletion, replicated across two rat strains, two independent labs (PMID:30622189 and PMID:38376248)","pmids":["30622189"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2019,"finding":"Deletion of GRA43 (or GRA42) causes GRA35 and other dense granule proteins to be retained inside the parasitophorous vacuole instead of localizing to the parasitophorous vacuole membrane, indicating that GRA43 mediates correct trafficking/localization of GRAs to the PVM.","method":"Knockout parasites (Δgra42, Δgra43) combined with fluorescence localization assays of GRA35 and other GRAs","journal":"mBio","confidence":"High","confidence_rationale":"Tier 2 / Moderate — clean KO with direct localization readout, two independent knockouts (Δgra42 and Δgra43) giving same phenotype, single lab","pmids":["30622189"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2019,"finding":"GRA43 (along with GRA35 and GRA42) is required for parasite in vivo fitness in the susceptible F344 rat strain independently of its role in NLRP1 inflammasome/pyroptosis activation, as Δgra43 parasites showed reduced cyst numbers in F344 rats.","method":"In vivo infection of F344 rats with Δgra43 parasites; brain cyst counting","journal":"mBio","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — clean KO with defined in vivo phenotype, but single lab and single method for the F344 fitness phenotype","pmids":["30622189"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2024,"finding":"Genome-wide CRISPR screen confirmed GRA43 (together with GRA35 and GRA42) as Toxoplasma effectors mediating cell death (pyroptosis) in Lewis rat macrophages. GRA43's role is upstream of GRA35's interaction with host ITCH E3 ubiquitin ligase at the PVM, consistent with its function in PVM localization of GRA35.","method":"Genome-wide CRISPR screen in Lewis rat macrophages","journal":"mBio","confidence":"High","confidence_rationale":"Tier 2 / Strong — genome-wide CRISPR screen with independent validation, replicates prior mutagenesis findings from PMID:30622189","pmids":["38376248"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2021,"finding":"GRA43 secreted by T. gondii triggers high levels of reactive oxygen species (ROS) production in iNOS-knockout SD rat peritoneal macrophages, which damages the parasitophorous vacuole membrane and mitochondrial membranes, leading to mitochondrial superoxide dismutase 2 depletion and macrophage pyroptosis/cell death.","method":"iNOS-knockout rat macrophage infection assays; ROS measurement; organelle membrane integrity assays; GRA43-attributed triggering by parasite secretion","journal":"iScience","confidence":"Medium","confidence_rationale":"Tier 3 / Weak — single lab, single study; mechanistic attribution of ROS production specifically to GRA43 relies on correlation with GRA43 secretion rather than direct GRA43 KO control in this system","pmids":["34765911"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2023,"finding":"GRA43 (as a PVM-targeting factor for dense granules) represses the CD8 T cell IFNγ differentiation response, suggesting that PVM-associated GRAs are important for CD8 T cell activation, as determined by genetic analysis of Δgra43 parasites in macrophage-CD8 T cell co-culture assays.","method":"Genetic mapping (QTL screen with F1 progeny), co-culture of Δgra43-infected macrophages with antigen-specific CD8 T cells (T57 transnuclear cells), IFNγ measurement","journal":"Frontiers in cellular and infection microbiology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, single method; GRA43's role described as a noted observation during ROCTR search rather than primary experimental focus","pmids":["37287466"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]}],"current_model":"GRA43 is a Toxoplasma gondii dense granule protein that localizes to the parasitophorous vacuole and is required for correct trafficking of GRA35 and other dense granule proteins to the parasitophorous vacuole membrane (PVM); through this PVM-organizing function it acts in the same pathway as GRA35 and GRA42 to enable GRA35-mediated activation of the NLRP1 inflammasome and pyroptosis in Lewis rat macrophages, and it also contributes to parasite fitness in vivo and represses host CD8 T cell IFNγ responses."},"narrative":{"mechanistic_narrative":"GRA43 is a Toxoplasma gondii dense granule protein that localizes to the parasitophorous vacuole and functions as a trafficking factor required for delivering GRA35 and other dense granule proteins to the parasitophorous vacuole membrane (PVM) [PMID:30622189]. Loss of GRA43 causes GRA35 and other GRAs to be retained inside the vacuole rather than reaching the PVM [PMID:30622189], and genetic epistasis places GRA43 in a common pathway with GRA35 and GRA42 [PMID:30622189]. Through this PVM-organizing role GRA43 acts upstream of GRA35's engagement of the host ITCH E3 ubiquitin ligase at the PVM to drive pyroptotic macrophage death in Lewis rats, such that GRA43 deletion strongly reduces cell death and enhances parasite replication [PMID:30622189, PMID:38376248]. GRA43 also contributes to parasite in vivo fitness in susceptible F344 rats independently of its inflammasome role [PMID:30622189].","teleology":[{"year":2019,"claim":"Establishing whether a specific parasite effector controls host macrophage death, this work identified GRA43 as a PV-localized dense granule protein individually required for Lewis rat macrophage pyroptosis and placed it in a shared pathway with GRA35 and GRA42.","evidence":"Chemical mutagenesis screen with whole-genome sequencing, CRISPR knockout, viability assays, and single/double/triple deletion epistasis","pmids":["30622189"],"confidence":"High","gaps":["Molecular mechanism by which GRA43 promotes trafficking not defined","No direct binding partners of GRA43 itself identified"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2019,"claim":"To explain how GRA43 enables downstream pyroptosis, deletion showed GRA43 is needed to traffic GRA35 and other dense granule proteins to the PVM rather than retaining them in the vacuole, defining its role as a PVM-targeting factor.","evidence":"Δgra42 and Δgra43 knockout parasites with fluorescence localization of GRA35 and other GRAs","pmids":["30622189"],"confidence":"High","gaps":["Whether GRA43 physically escorts cargo or organizes a translocation machinery is unknown","Full set of GRA43-dependent cargo not enumerated"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2019,"claim":"To separate GRA43's inflammasome role from other functions, in vivo infection revealed GRA43 is required for parasite fitness in susceptible F344 rats independently of NLRP1/pyroptosis activation.","evidence":"In vivo infection of F344 rats with Δgra43 parasites and brain cyst counting","pmids":["30622189"],"confidence":"Medium","gaps":["Single lab and single method for the F344 fitness phenotype","Mechanistic basis of the fitness defect not resolved"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2021,"claim":"Probing how effector secretion converts to macrophage death, GRA43 secretion was linked to ROS induction that damages PVM and mitochondrial membranes, depleting SOD2 and driving pyroptosis in iNOS-knockout rat macrophages.","evidence":"iNOS-knockout rat macrophage infection, ROS measurement, and organelle membrane integrity assays","pmids":["34765911"],"confidence":"Medium","gaps":["ROS attribution relies on correlation with GRA43 secretion rather than a direct Δgra43 control in this system","How ROS mechanistically connects to the GRA35/GRA42 pathway is unclear"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2023,"claim":"Extending GRA43's immune impact beyond pyroptosis, genetic analysis indicated GRA43 represses the CD8 T cell IFNγ differentiation response, implicating PVM-associated GRAs in modulating CD8 T cell activation.","evidence":"QTL mapping with F1 progeny and co-culture of Δgra43-infected macrophages with antigen-specific CD8 T cells with IFNγ readout","pmids":["37287466"],"confidence":"Low","gaps":["Observed as a secondary note rather than primary experimental focus; single lab and single method","Direct mechanism linking GRA43-dependent PVM localization to CD8 T cell suppression not defined"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2024,"claim":"Independently confirming and ordering the pathway, a genome-wide CRISPR screen validated GRA43 (with GRA35 and GRA42) as a cell-death effector acting upstream of GRA35's interaction with host ITCH at the PVM.","evidence":"Genome-wide CRISPR screen in Lewis rat macrophages","pmids":["38376248"],"confidence":"High","gaps":["Direct biochemical interaction between GRA43 and any host or parasite factor still not shown","Structural basis of GRA43 trafficking function unresolved"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":null,"claim":"The molecular mechanism by which GRA43 directs dense granule proteins to the PVM — its direct binding partners, cargo recognition, and biochemical activity — remains uncharacterized in the available corpus.","evidence":"","pmids":[],"confidence":"Low","gaps":["No direct GRA43 interactome","No structural model","Mechanism of cargo selection and PVM targeting unknown"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]}],"mechanism_profile":{"molecular_activity":[],"localization":[],"pathway":[],"complexes":[],"partners":["GRA35 (DENSE GRANULE PROTEIN TRAFFICKED TO PVM IN A GRA43-DEPENDENT MANNER) [#1, #3]","GRA42 (ACTS IN THE SAME PVM-TRAFFICKING PATHWAY) [#0, #1]"],"other_free_text":["GRA43 acts upstream of GRA35's engagement of host ITCH E3 ubiquitin ligase at the PVM [#3].","GRA43 deletion reduces macrophage pyroptosis and enhances parasite replication [#0].","GRA43 contributes to parasite fitness in F344 rats independently of the inflammasome role [#2]."]},"strain_conflicts":[]},"prefetch_data":{},"corpus_meta":[{"pmid":"30622189","id":"PMC_30622189","title":"Three Toxoplasma gondii Dense Granule Proteins Are Required for Induction of Lewis Rat Macrophage Pyroptosis.","date":"2019","source":"mBio","url":"https://pubmed.ncbi.nlm.nih.gov/30622189","citation_count":63,"is_preprint":false},{"pmid":"34765911","id":"PMC_34765911","title":"High resistance to Toxoplasma gondii infection in inducible nitric oxide synthase knockout rats.","date":"2021","source":"iScience","url":"https://pubmed.ncbi.nlm.nih.gov/34765911","citation_count":14,"is_preprint":false},{"pmid":"38029261","id":"PMC_38029261","title":"Protective immunity induced by DNA vaccine containing TgGRA35, TgGRA42, and TgGRA43 against Toxoplasma gondii infection in Kunming mice.","date":"2023","source":"Frontiers in cellular and infection microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/38029261","citation_count":13,"is_preprint":false},{"pmid":"38376248","id":"PMC_38376248","title":"Host E3 ubiquitin ligase ITCH mediates Toxoplasma gondii effector GRA35-triggered NLRP1 inflammasome activation and cell-autonomous immunity.","date":"2024","source":"mBio","url":"https://pubmed.ncbi.nlm.nih.gov/38376248","citation_count":11,"is_preprint":false},{"pmid":"37287466","id":"PMC_37287466","title":"Variation in CD8 T cell IFNγ differentiation to strains of Toxoplasma gondii is characterized by small effect QTLs with contribution from ROP16.","date":"2023","source":"Frontiers in cellular and infection microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/37287466","citation_count":1,"is_preprint":false},{"pmid":"38168400","id":"PMC_38168400","title":"Host E3 ubiquitin ligase ITCH mediates Toxoplasma gondii effector GRA35-triggered NLRP1 inflammasome activation and cell-autonomous immunity.","date":"2023","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/38168400","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":6338,"output_tokens":1748,"usd":0.022617,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":6964,"output_tokens":2895,"usd":0.053597,"stage2_stop_reason":"end_turn"},"total_usd":0.0,"stage1_batch_id":"msgbatch_01XFXJD6iZecXyWn9uU5Qom9","stage2_batch_id":"msgbatch_013GToKgs4AKSCRE4yvueviH","note":"batch pricing = 50% of standard","skipped":true},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2019,\n      \"finding\": \"GRA43 is a parasitophorous vacuole-localized dense granule protein individually required for induction of Lewis rat macrophage pyroptosis; deletion of GRA43 greatly reduces macrophage cell death and enhances parasite replication. Epistasis analysis (single, double, and triple deletion mutants showing similar viability levels) places GRA35, GRA42, and GRA43 in the same pathway.\",\n      \"method\": \"Chemical mutagenesis screen, whole-genome sequencing, CRISPR/knockout (Δgra43), cell viability assays, in vivo infection of Lewis rats\",\n      \"journal\": \"mBio\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean KO with defined cellular phenotype, genetic epistasis via single/double/triple deletion, replicated across two rat strains, two independent labs (PMID:30622189 and PMID:38376248)\",\n      \"pmids\": [\"30622189\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Deletion of GRA43 (or GRA42) causes GRA35 and other dense granule proteins to be retained inside the parasitophorous vacuole instead of localizing to the parasitophorous vacuole membrane, indicating that GRA43 mediates correct trafficking/localization of GRAs to the PVM.\",\n      \"method\": \"Knockout parasites (Δgra42, Δgra43) combined with fluorescence localization assays of GRA35 and other GRAs\",\n      \"journal\": \"mBio\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean KO with direct localization readout, two independent knockouts (Δgra42 and Δgra43) giving same phenotype, single lab\",\n      \"pmids\": [\"30622189\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"GRA43 (along with GRA35 and GRA42) is required for parasite in vivo fitness in the susceptible F344 rat strain independently of its role in NLRP1 inflammasome/pyroptosis activation, as Δgra43 parasites showed reduced cyst numbers in F344 rats.\",\n      \"method\": \"In vivo infection of F344 rats with Δgra43 parasites; brain cyst counting\",\n      \"journal\": \"mBio\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — clean KO with defined in vivo phenotype, but single lab and single method for the F344 fitness phenotype\",\n      \"pmids\": [\"30622189\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Genome-wide CRISPR screen confirmed GRA43 (together with GRA35 and GRA42) as Toxoplasma effectors mediating cell death (pyroptosis) in Lewis rat macrophages. GRA43's role is upstream of GRA35's interaction with host ITCH E3 ubiquitin ligase at the PVM, consistent with its function in PVM localization of GRA35.\",\n      \"method\": \"Genome-wide CRISPR screen in Lewis rat macrophages\",\n      \"journal\": \"mBio\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genome-wide CRISPR screen with independent validation, replicates prior mutagenesis findings from PMID:30622189\",\n      \"pmids\": [\"38376248\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"GRA43 secreted by T. gondii triggers high levels of reactive oxygen species (ROS) production in iNOS-knockout SD rat peritoneal macrophages, which damages the parasitophorous vacuole membrane and mitochondrial membranes, leading to mitochondrial superoxide dismutase 2 depletion and macrophage pyroptosis/cell death.\",\n      \"method\": \"iNOS-knockout rat macrophage infection assays; ROS measurement; organelle membrane integrity assays; GRA43-attributed triggering by parasite secretion\",\n      \"journal\": \"iScience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, single study; mechanistic attribution of ROS production specifically to GRA43 relies on correlation with GRA43 secretion rather than direct GRA43 KO control in this system\",\n      \"pmids\": [\"34765911\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"GRA43 (as a PVM-targeting factor for dense granules) represses the CD8 T cell IFNγ differentiation response, suggesting that PVM-associated GRAs are important for CD8 T cell activation, as determined by genetic analysis of Δgra43 parasites in macrophage-CD8 T cell co-culture assays.\",\n      \"method\": \"Genetic mapping (QTL screen with F1 progeny), co-culture of Δgra43-infected macrophages with antigen-specific CD8 T cells (T57 transnuclear cells), IFNγ measurement\",\n      \"journal\": \"Frontiers in cellular and infection microbiology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, single method; GRA43's role described as a noted observation during ROCTR search rather than primary experimental focus\",\n      \"pmids\": [\"37287466\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    }\n  ],\n  \"current_model\": \"GRA43 is a Toxoplasma gondii dense granule protein that localizes to the parasitophorous vacuole and is required for correct trafficking of GRA35 and other dense granule proteins to the parasitophorous vacuole membrane (PVM); through this PVM-organizing function it acts in the same pathway as GRA35 and GRA42 to enable GRA35-mediated activation of the NLRP1 inflammasome and pyroptosis in Lewis rat macrophages, and it also contributes to parasite fitness in vivo and represses host CD8 T cell IFNγ responses.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"GRA43 is a Toxoplasma gondii dense granule protein that localizes to the parasitophorous vacuole and functions as a trafficking factor required for delivering GRA35 and other dense granule proteins to the parasitophorous vacuole membrane (PVM) [#0, #1]. Loss of GRA43 causes GRA35 and other GRAs to be retained inside the vacuole rather than reaching the PVM [#1], and genetic epistasis places GRA43 in a common pathway with GRA35 and GRA42 [#0]. Through this PVM-organizing role GRA43 acts upstream of GRA35's engagement of the host ITCH E3 ubiquitin ligase at the PVM to drive pyroptotic macrophage death in Lewis rats, such that GRA43 deletion strongly reduces cell death and enhances parasite replication [#0, #3]. GRA43 also contributes to parasite in vivo fitness in susceptible F344 rats independently of its inflammasome role [#2].\",\n  \"teleology\": [\n    {\n      \"year\": 2019,\n      \"claim\": \"Establishing whether a specific parasite effector controls host macrophage death, this work identified GRA43 as a PV-localized dense granule protein individually required for Lewis rat macrophage pyroptosis and placed it in a shared pathway with GRA35 and GRA42.\",\n      \"evidence\": \"Chemical mutagenesis screen with whole-genome sequencing, CRISPR knockout, viability assays, and single/double/triple deletion epistasis\",\n      \"pmids\": [\"30622189\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Molecular mechanism by which GRA43 promotes trafficking not defined\",\n        \"No direct binding partners of GRA43 itself identified\"\n      ],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"To explain how GRA43 enables downstream pyroptosis, deletion showed GRA43 is needed to traffic GRA35 and other dense granule proteins to the PVM rather than retaining them in the vacuole, defining its role as a PVM-targeting factor.\",\n      \"evidence\": \"Δgra42 and Δgra43 knockout parasites with fluorescence localization of GRA35 and other GRAs\",\n      \"pmids\": [\"30622189\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether GRA43 physically escorts cargo or organizes a translocation machinery is unknown\",\n        \"Full set of GRA43-dependent cargo not enumerated\"\n      ],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"To separate GRA43's inflammasome role from other functions, in vivo infection revealed GRA43 is required for parasite fitness in susceptible F344 rats independently of NLRP1/pyroptosis activation.\",\n      \"evidence\": \"In vivo infection of F344 rats with Δgra43 parasites and brain cyst counting\",\n      \"pmids\": [\"30622189\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Single lab and single method for the F344 fitness phenotype\",\n        \"Mechanistic basis of the fitness defect not resolved\"\n      ],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Probing how effector secretion converts to macrophage death, GRA43 secretion was linked to ROS induction that damages PVM and mitochondrial membranes, depleting SOD2 and driving pyroptosis in iNOS-knockout rat macrophages.\",\n      \"evidence\": \"iNOS-knockout rat macrophage infection, ROS measurement, and organelle membrane integrity assays\",\n      \"pmids\": [\"34765911\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"ROS attribution relies on correlation with GRA43 secretion rather than a direct Δgra43 control in this system\",\n        \"How ROS mechanistically connects to the GRA35/GRA42 pathway is unclear\"\n      ],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Extending GRA43's immune impact beyond pyroptosis, genetic analysis indicated GRA43 represses the CD8 T cell IFNγ differentiation response, implicating PVM-associated GRAs in modulating CD8 T cell activation.\",\n      \"evidence\": \"QTL mapping with F1 progeny and co-culture of Δgra43-infected macrophages with antigen-specific CD8 T cells with IFNγ readout\",\n      \"pmids\": [\"37287466\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"Observed as a secondary note rather than primary experimental focus; single lab and single method\",\n        \"Direct mechanism linking GRA43-dependent PVM localization to CD8 T cell suppression not defined\"\n      ],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Independently confirming and ordering the pathway, a genome-wide CRISPR screen validated GRA43 (with GRA35 and GRA42) as a cell-death effector acting upstream of GRA35's interaction with host ITCH at the PVM.\",\n      \"evidence\": \"Genome-wide CRISPR screen in Lewis rat macrophages\",\n      \"pmids\": [\"38376248\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Direct biochemical interaction between GRA43 and any host or parasite factor still not shown\",\n        \"Structural basis of GRA43 trafficking function unresolved\"\n      ],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The molecular mechanism by which GRA43 directs dense granule proteins to the PVM — its direct binding partners, cargo recognition, and biochemical activity — remains uncharacterized in the available corpus.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No direct GRA43 interactome\",\n        \"No structural model\",\n        \"Mechanism of cargo selection and PVM targeting unknown\"\n      ],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140318\", \"supporting_discovery_ids\": [1], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0020005\", \"supporting_discovery_ids\": [0, 1], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]},\n      {\"term_id\": \"GO:0020003\", \"supporting_discovery_ids\": [0], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0070269\", \"supporting_discovery_ids\": [0, 3], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]},\n      {\"term_id\": \"GO:0052167\", \"supporting_discovery_ids\": [5], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"GRA35 (dense granule protein trafficked to PVM in a GRA43-dependent manner) [#1, #3]\",\n      \"GRA42 (acts in the same PVM-trafficking pathway) [#0, #1]\"\n    ],\n    \"other_free_text\": [\n      \"GRA43 acts upstream of GRA35's engagement of host ITCH E3 ubiquitin ligase at the PVM [#3].\",\n      \"GRA43 deletion reduces macrophage pyroptosis and enhances parasite replication [#0].\",\n      \"GRA43 contributes to parasite fitness in F344 rats independently of the inflammasome role [#2].\"\n    ]\n  },\n  \"strain_conflicts\": []\n}\n```","audit_flag":null,"strains":[{"strain":"ME49","gene_id":"TGME49_237015","name":"GRA43","product":"dense granule protein GRA43","uniprot_acc":"S8ET88","chromosome":"TGME49_chrX","start_pos":5673387},{"strain":"GT1","gene_id":"TGGT1_237015","name":null,"product":"hypothetical protein","uniprot_acc":null,"chromosome":"TGGT1_chrX","start_pos":5566838},{"strain":"RH88","gene_id":"TGRH88_040910","name":null,"product":"unspecified product","uniprot_acc":null,"chromosome":"CM023091","start_pos":1860765},{"strain":"VEG","gene_id":"TGVEG_237015","name":null,"product":"putative transmembrane protein","uniprot_acc":"B9Q6H9","chromosome":"TGVEG_chrX","start_pos":5600651}]}