{"gene":"TG_249990","gene_symbol":"GRA70","run_date":"2026-09-08T15:31:38+00:00","timeline":{"discoveries":[{"year":2023,"finding":"GRA70 is a dense granule protein that co-immunoprecipitates with GRA32 and two other GRA32 homologs, suggesting formation of a multi-protein complex; deletion of individual members leads to early parasite egress in IFNγ-stimulated human fibroblasts.","method":"Co-immunoprecipitation, gene knockout, growth assay in IFNγ-stimulated primary human fibroblasts","journal":"mBio","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP and KO phenotype in single study, single lab, two orthogonal methods","pmids":["36916910"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2023,"finding":"GRA70 forms a heterotrimeric complex with GRA57 and GRA71 that enhances parasite survival in IFNγ-activated human foreskin fibroblasts; knockout of GRA70 reduces parasite persistence in IFNγ-activated human cells.","method":"Targeted CRISPR screen (two Toxoplasma strains), gene knockout, survival assay in IFNγ-activated human foreskin fibroblasts","journal":"PLoS biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — CRISPR screen validated by KO in two strains, single lab, two orthogonal approaches (screen + KO phenotype)","pmids":["37459303"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2023,"finding":"Vacuoles from GRA70 knockout parasites display reduced host-mediated ubiquitination compared to wild-type, but this reduced ubiquitination does not account for the IFNγ resistance phenotype mediated by the GRA57/GRA70/GRA71 complex (i.e., ubiquitination change is a secondary consequence of complex deletion).","method":"Gene knockout, ubiquitination assay of parasite vacuoles in IFNγ-activated human cells","journal":"PLoS biology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, single method, and authors note this is likely a secondary (negative/dissociative) result","pmids":["37459303"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]}],"current_model":"GRA70 is a Toxoplasma dense granule effector protein that forms a heterotrimeric complex with GRA57 and GRA71 (and associates with GRA32); this complex is required for parasite survival in IFNγ-activated human cells, at least in part by preventing premature parasite egress, independently of host-cell ubiquitination of the parasitophorous vacuole."},"narrative":{"mechanistic_narrative":"GRA70 is a Toxoplasma gondii dense granule effector that supports parasite survival within IFNγ-activated human cells [PMID:36916910, PMID:37459303]. It assembles into a multi-protein complex, forming a heterotrimeric complex with GRA57 and GRA71 and co-immunoprecipitating with GRA32 and additional GRA32 homologs [PMID:36916910, PMID:37459303]. Loss of GRA70 causes premature parasite egress from IFNγ-stimulated human fibroblasts and reduces parasite persistence in IFNγ-activated cells, establishing the complex as a determinant of intracellular survival under host restriction [PMID:36916910, PMID:37459303]. GRA70-knockout vacuoles show reduced host-mediated ubiquitination, but this change does not account for the IFNγ resistance phenotype and represents a secondary consequence of complex disruption [PMID:37459303]. Beyond these findings, the molecular activity by which GRA70 prevents egress and confers IFNγ resistance has not been characterized in the available corpus.","teleology":[{"year":2023,"claim":"Establishing that GRA70 is a dense granule protein embedded in a multi-protein complex answered where it acts and with which partners, framing it as a candidate effector rather than an isolated protein.","evidence":"Reciprocal co-immunoprecipitation and gene knockout with growth assays in IFNγ-stimulated primary human fibroblasts","pmids":["36916910"],"confidence":"Medium","gaps":["The stoichiometry and molecular architecture of the GRA70/GRA32-homolog association are not resolved","The mechanism by which complex loss triggers early egress is unknown","No enzymatic or biochemical activity assigned to GRA70"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2023,"claim":"Defining GRA70 as part of a heterotrimeric GRA57/GRA70/GRA71 complex required for survival identified the functional unit and its role in resisting IFNγ-mediated host restriction.","evidence":"Targeted CRISPR screen across two Toxoplasma strains with knockout survival assays in IFNγ-activated human foreskin fibroblasts","pmids":["37459303"],"confidence":"Medium","gaps":["The host target(s) or pathway the complex neutralizes is unidentified","Whether GRA70 contributes catalytic versus structural function within the complex is unknown","The relationship between the GRA57/GRA70/GRA71 heterotrimer and the GRA32-homolog association is not defined"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2023,"claim":"Testing whether altered vacuolar ubiquitination explains the phenotype ruled out this mechanism, showing the IFNγ resistance conferred by the complex is independent of host-mediated ubiquitination of the vacuole.","evidence":"Gene knockout and ubiquitination assay of parasite vacuoles in IFNγ-activated human cells","pmids":["37459303"],"confidence":"Low","gaps":["Single-lab, single-method dissociative result interpreted as secondary; the causal mechanism of IFNγ resistance remains unidentified","The direct molecular effector activity preventing premature egress is not established"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":null,"claim":"The molecular mechanism by which the GRA57/GRA70/GRA71 complex prevents premature egress and confers IFNγ resistance remains unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["No host target or biochemical activity has been assigned to GRA70","How the complex mechanistically stabilizes the parasite against IFNγ-driven egress is unknown"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]}],"mechanism_profile":{"molecular_activity":[],"localization":[],"pathway":[],"complexes":["Heterotrimeric GRA57/GRA70/GRA71 complex [#1]","GRA70 association with GRA32 and additional GRA32 homologs [#0]"],"partners":["GRA57 [#1]","GRA71 [#1]","GRA32 [#0]","GRA32 HOMOLOGS [#0]"],"other_free_text":["Dense granule protein of Toxoplasma gondii [#0]","Required for parasite survival and persistence in IFNγ-activated human fibroblasts [#0, #1]","Loss causes premature parasite egress in IFNγ-stimulated human cells [#0]","IFNγ resistance phenotype is independent of host-mediated vacuolar ubiquitination [#2]"]},"strain_conflicts":[]},"prefetch_data":{},"corpus_meta":[{"pmid":"36916910","id":"PMC_36916910","title":"CRISPR Screens Identify Toxoplasma Genes That Determine Parasite Fitness in Interferon Gamma-Stimulated Human Cells.","date":"2023","source":"mBio","url":"https://pubmed.ncbi.nlm.nih.gov/36916910","citation_count":29,"is_preprint":false},{"pmid":"37459303","id":"PMC_37459303","title":"A heterotrimeric complex of Toxoplasma proteins promotes parasite survival in interferon gamma-stimulated human cells.","date":"2023","source":"PLoS biology","url":"https://pubmed.ncbi.nlm.nih.gov/37459303","citation_count":24,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":3545,"output_tokens":840,"usd":0.011617,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":5654,"output_tokens":1953,"usd":0.038547,"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\": 2023,\n      \"finding\": \"GRA70 is a dense granule protein that co-immunoprecipitates with GRA32 and two other GRA32 homologs, suggesting formation of a multi-protein complex; deletion of individual members leads to early parasite egress in IFNγ-stimulated human fibroblasts.\",\n      \"method\": \"Co-immunoprecipitation, gene knockout, growth assay in IFNγ-stimulated primary human fibroblasts\",\n      \"journal\": \"mBio\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP and KO phenotype in single study, single lab, two orthogonal methods\",\n      \"pmids\": [\"36916910\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"GRA70 forms a heterotrimeric complex with GRA57 and GRA71 that enhances parasite survival in IFNγ-activated human foreskin fibroblasts; knockout of GRA70 reduces parasite persistence in IFNγ-activated human cells.\",\n      \"method\": \"Targeted CRISPR screen (two Toxoplasma strains), gene knockout, survival assay in IFNγ-activated human foreskin fibroblasts\",\n      \"journal\": \"PLoS biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — CRISPR screen validated by KO in two strains, single lab, two orthogonal approaches (screen + KO phenotype)\",\n      \"pmids\": [\"37459303\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Vacuoles from GRA70 knockout parasites display reduced host-mediated ubiquitination compared to wild-type, but this reduced ubiquitination does not account for the IFNγ resistance phenotype mediated by the GRA57/GRA70/GRA71 complex (i.e., ubiquitination change is a secondary consequence of complex deletion).\",\n      \"method\": \"Gene knockout, ubiquitination assay of parasite vacuoles in IFNγ-activated human cells\",\n      \"journal\": \"PLoS biology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, single method, and authors note this is likely a secondary (negative/dissociative) result\",\n      \"pmids\": [\"37459303\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    }\n  ],\n  \"current_model\": \"GRA70 is a Toxoplasma dense granule effector protein that forms a heterotrimeric complex with GRA57 and GRA71 (and associates with GRA32); this complex is required for parasite survival in IFNγ-activated human cells, at least in part by preventing premature parasite egress, independently of host-cell ubiquitination of the parasitophorous vacuole.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"GRA70 is a Toxoplasma gondii dense granule effector that supports parasite survival within IFN\\u03b3-activated human cells [#0, #1]. It assembles into a multi-protein complex, forming a heterotrimeric complex with GRA57 and GRA71 and co-immunoprecipitating with GRA32 and additional GRA32 homologs [#0, #1]. Loss of GRA70 causes premature parasite egress from IFN\\u03b3-stimulated human fibroblasts and reduces parasite persistence in IFN\\u03b3-activated cells, establishing the complex as a determinant of intracellular survival under host restriction [#0, #1]. GRA70-knockout vacuoles show reduced host-mediated ubiquitination, but this change does not account for the IFN\\u03b3 resistance phenotype and represents a secondary consequence of complex disruption [#2]. Beyond these findings, the molecular activity by which GRA70 prevents egress and confers IFN\\u03b3 resistance has not been characterized in the available corpus.\",\n  \"teleology\": [\n    {\n      \"year\": 2023,\n      \"claim\": \"Establishing that GRA70 is a dense granule protein embedded in a multi-protein complex answered where it acts and with which partners, framing it as a candidate effector rather than an isolated protein.\",\n      \"evidence\": \"Reciprocal co-immunoprecipitation and gene knockout with growth assays in IFN\\u03b3-stimulated primary human fibroblasts\",\n      \"pmids\": [\n        \"36916910\"\n      ],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"The stoichiometry and molecular architecture of the GRA70/GRA32-homolog association are not resolved\",\n        \"The mechanism by which complex loss triggers early egress is unknown\",\n        \"No enzymatic or biochemical activity assigned to GRA70\"\n      ],\n      \"attribution\": [\n        {\n          \"species\": \"Toxoplasma gondii\",\n          \"strain\": \"\"\n        }\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Defining GRA70 as part of a heterotrimeric GRA57/GRA70/GRA71 complex required for survival identified the functional unit and its role in resisting IFN\\u03b3-mediated host restriction.\",\n      \"evidence\": \"Targeted CRISPR screen across two Toxoplasma strains with knockout survival assays in IFN\\u03b3-activated human foreskin fibroblasts\",\n      \"pmids\": [\n        \"37459303\"\n      ],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"The host target(s) or pathway the complex neutralizes is unidentified\",\n        \"Whether GRA70 contributes catalytic versus structural function within the complex is unknown\",\n        \"The relationship between the GRA57/GRA70/GRA71 heterotrimer and the GRA32-homolog association is not defined\"\n      ],\n      \"attribution\": [\n        {\n          \"species\": \"Toxoplasma gondii\",\n          \"strain\": \"\"\n        }\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Testing whether altered vacuolar ubiquitination explains the phenotype ruled out this mechanism, showing the IFN\\u03b3 resistance conferred by the complex is independent of host-mediated ubiquitination of the vacuole.\",\n      \"evidence\": \"Gene knockout and ubiquitination assay of parasite vacuoles in IFN\\u03b3-activated human cells\",\n      \"pmids\": [\n        \"37459303\"\n      ],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"Single-lab, single-method dissociative result interpreted as secondary; the causal mechanism of IFN\\u03b3 resistance remains unidentified\",\n        \"The direct molecular effector activity preventing premature egress is not established\"\n      ],\n      \"attribution\": [\n        {\n          \"species\": \"Toxoplasma gondii\",\n          \"strain\": \"\"\n        }\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The molecular mechanism by which the GRA57/GRA70/GRA71 complex prevents premature egress and confers IFN\\u03b3 resistance remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No host target or biochemical activity has been assigned to GRA70\",\n        \"How the complex mechanistically stabilizes the parasite against IFN\\u03b3-driven egress is unknown\"\n      ],\n      \"attribution\": [\n        {\n          \"species\": \"Toxoplasma gondii\",\n          \"strain\": \"\"\n        }\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [],\n    \"localization\": [],\n    \"pathway\": [],\n    \"complexes\": [\n      \"Heterotrimeric GRA57/GRA70/GRA71 complex [#1]\",\n      \"GRA70 association with GRA32 and additional GRA32 homologs [#0]\"\n    ],\n    \"partners\": [\n      \"GRA57 [#1]\",\n      \"GRA71 [#1]\",\n      \"GRA32 [#0]\",\n      \"GRA32 homologs [#0]\"\n    ],\n    \"other_free_text\": [\n      \"Dense granule protein of Toxoplasma gondii [#0]\",\n      \"Required for parasite survival and persistence in IFN\\u03b3-activated human fibroblasts [#0, #1]\",\n      \"Loss causes premature parasite egress in IFN\\u03b3-stimulated human cells [#0]\",\n      \"IFN\\u03b3 resistance phenotype is independent of host-mediated vacuolar ubiquitination [#2]\"\n    ]\n  },\n  \"strain_conflicts\": []\n}\n```","audit_flag":null,"strains":[{"strain":"ME49","gene_id":"TGME49_249990","name":"GRA70","product":"dense granule protein GRA70","uniprot_acc":"A0A125YQ73","chromosome":"TGME49_chrXII","start_pos":4972033},{"strain":"GT1","gene_id":"TGGT1_249990","name":null,"product":"hypothetical protein","uniprot_acc":null,"chromosome":"TGGT1_chrXII","start_pos":4909167},{"strain":"RH88","gene_id":"TGRH88_063740","name":null,"product":"unspecified product","uniprot_acc":null,"chromosome":"CM023093","start_pos":5031133},{"strain":"VEG","gene_id":"TGVEG_249990","name":null,"product":"putative microtubule-binding protein","uniprot_acc":"B6KHK8","chromosome":"TGVEG_chrXII","start_pos":4948353}]}