{"gene":"TG_321300","gene_symbol":"ATG12","run_date":"2026-09-08T15:15:14","timeline":{"discoveries":[{"year":2019,"finding":"In Toxoplasma gondii (and Plasmodium), ATG12 lacks the C-terminal glycine required for covalent conjugation and the E2-like enzyme ATG10 is absent; instead, TgATG12 forms a non-covalent complex with ATG5. This non-covalent ATG12-ATG5 complex retains the ability to facilitate ATG8-phosphatidylethanolamine conjugation, demonstrating an evolutionary transition from covalent conjugation to non-covalent interaction in the ATG12 system.","method":"Biochemical reconstitution, sequence/structural analysis, in vitro ATG8 lipidation assay","journal":"Nature structural & molecular biology","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution of non-covalent complex with functional ATG8 lipidation assay, supported by sequence analysis across multiple organisms","pmids":["30911187"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""},{"species":"Plasmodium","strain":""}]},{"year":2022,"finding":"TgATG12 forms a complex with TgATG5 and TgATG16L in Toxoplasma gondii. This TgATG12-TgATG5-TgATG16L complex localizes to isolated structures at the periphery of the apicoplast in a TgATG16L-dependent manner. Inducible depletion of TgATG12 caused loss of the apicoplast and impaired parasite growth, establishing a required role for TgATG12 in apicoplast biogenesis.","method":"Co-immunoprecipitation, inducible auxin-degron knockdown, fluorescence microscopy, parasite growth assay","journal":"Autophagy","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP, inducible depletion with specific apicoplast phenotype, multiple orthogonal methods in one study","pmids":["36095096"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2022,"finding":"The TgATG12-TgATG5-TgATG16L complex is required for TgATG8 localization to the apicoplast. A putative ubiquitin-interacting motif-docking site (UDS) on TgATG8 was identified; mutation of this UDS abolished TgATG8 localization on the apicoplast without affecting ATG8 lipidation, and a SNARE protein TgSNAP29 was required for TgATG8 vesicular translocation to the apicoplast.","method":"Site-directed mutagenesis of UDS, inducible knockdown (AID), fluorescence microscopy, lipidation assay","journal":"Autophagy","confidence":"High","confidence_rationale":"Tier 1 / Moderate — mutagenesis of functional site combined with inducible depletion and multiple readouts in one study","pmids":["36095096"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2014,"finding":"The host ATG12-ATG5-ATG16L1 complex (together with ATG7 and ATG3) is required for targeting LC3 and IFN-γ effectors to the parasitophorous vacuole membrane (PVM) of T. gondii and for subsequent disruption of the PVM. Pharmacological modulation of degradative autophagy or deletion of other essential autophagy genes had no substantial effect, indicating specificity of the LC3 conjugation system in this immune function.","method":"Genetic knockout of host autophagy genes, in vitro infection assay, in vivo mouse model, immunofluorescence microscopy","journal":"Immunity","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean genetic knockouts in multiple autophagy components with specific mechanistic readout (LC3/effector targeting to PVM), replicated in vitro and in vivo","pmids":["24931121"],"is_preprint":false,"attribution":[{"species":"Mus musculus","strain":""}]},{"year":2016,"finding":"The host ATG12-ATG5-ATG16L1 complex specifies the conjugation site of LC3 homologs (LC3, GABARAP, GABARAPL1, GABARAPL2) on the PVM of T. gondii. Experimental relocation of the ATG12-ATG5-ATG16L1 complex to alternative membranes redirected IFN-γ-inducible GTPase targeting to those new membranes rather than the PVM, demonstrating that ATG12-ATG5-ATG16L1 complex localization is necessary and sufficient to determine the membrane destination of LC3 homologs and downstream immune effectors.","method":"Knockout/knockdown of LC3 homologs, artificial membrane targeting of ATG12-ATG5-ATG16L1 complex, immunofluorescence, infection assay","journal":"Autophagy","confidence":"High","confidence_rationale":"Tier 2 / Strong — gain-of-function membrane retargeting experiment combined with loss-of-function genetics, multiple orthogonal methods, mechanistically definitive","pmids":["27172324"],"is_preprint":false,"attribution":[{"species":"Mus musculus","strain":""}]}],"current_model":"In Toxoplasma gondii, ATG12 lacks the canonical C-terminal glycine and the E2-like enzyme ATG10, so it forms a non-covalent (rather than covalent) complex with ATG5; this TgATG12–TgATG5–TgATG16L complex localizes to the apicoplast periphery, is essential for apicoplast biogenesis and parasite viability, and facilitates TgATG8–phosphatidylethanolamine conjugation; in the host cell, the orthologous ATG12–ATG5–ATG16L1 complex specifies where LC3 homologs are conjugated to membranes and thereby directs IFN-γ immune effectors to the parasitophorous vacuole membrane for its disruption."},"narrative":{"mechanistic_narrative":"ATG12 is a core component of the ATG12–ATG5–ATG16L conjugation machinery that specifies where ATG8/LC3-family proteins are conjugated to membranes, a function conserved from apicomplexan parasites to their mammalian hosts [PMID:30911187, PMID:36095096, PMID:24931121]. In Toxoplasma gondii, ATG12 has diverged from the canonical ubiquitin-like pathway: it lacks the C-terminal glycine required for covalent conjugation and its cognate E2-like enzyme ATG10 is absent, so TgATG12 instead forms a non-covalent complex with ATG5 that nonetheless retains the ability to promote ATG8–phosphatidylethanolamine conjugation [PMID:30911187]. This TgATG12–TgATG5–TgATG16L complex localizes to the apicoplast periphery in a TgATG16L-dependent manner and is required for apicoplast biogenesis and parasite growth [PMID:36095096], directing TgATG8 to the apicoplast via a ubiquitin-interacting-motif-docking site on TgATG8 and SNAP29-dependent vesicular translocation [PMID:36095096]. In the mammalian host, the orthologous ATG12–ATG5–ATG16L1 complex is required to target LC3 homologs and IFN-γ effectors to the T. gondii parasitophorous vacuole membrane (PVM) and disrupt it [PMID:24931121], and the complex's own membrane localization is both necessary and sufficient to determine where LC3 homologs and downstream IFN-γ-inducible GTPases are deposited [PMID:27172324].","teleology":[{"year":2014,"claim":"It was unknown which autophagy machinery directs anti-Toxoplasma IFN-γ immunity; genetic dissection showed the LC3 conjugation system, not degradative autophagy, targets LC3 and IFN-γ effectors to the PVM for its disruption, establishing a non-canonical immune role for ATG12–ATG5–ATG16L1.","evidence":"Genetic knockout of host autophagy genes with in vitro infection and in vivo mouse models plus immunofluorescence","pmids":["24931121"],"confidence":"High","gaps":["Did not resolve how the complex is recruited specifically to the PVM","Did not distinguish the roles of individual complex members"],"attribution":[{"species":"Mus musculus","strain":""}]},{"year":2016,"claim":"Whether ATG12–ATG5–ATG16L1 merely conjugates LC3 or dictates its destination was unresolved; retargeting the complex to alternative membranes redirected LC3 homologs and IFN-γ GTPases there, establishing that complex localization is necessary and sufficient to specify the conjugation site.","evidence":"Loss-of-function knockdown of LC3 homologs combined with gain-of-function artificial membrane targeting, immunofluorescence, and infection assays","pmids":["27172324"],"confidence":"High","gaps":["Did not identify the receptor that positions the complex on native target membranes","Restricted to the host anti-parasite context"],"attribution":[{"species":"Mus musculus","strain":""}]},{"year":2019,"claim":"It was unclear whether ATG12 function requires covalent conjugation in all lineages; reconstitution showed T. gondii and Plasmodium ATG12 lack the C-terminal glycine and ATG10, forming a non-covalent ATG12–ATG5 complex that still supports ATG8 lipidation, revealing an evolutionary shift from covalent to non-covalent assembly.","evidence":"Biochemical reconstitution, sequence/structural analysis, and in vitro ATG8 lipidation assay","pmids":["30911187"],"confidence":"High","gaps":["Did not establish the in-cell consequences of the non-covalent complex","Structural basis of the non-covalent interaction not resolved"],"attribution":[{"species":"Toxoplasma gondii","strain":""},{"species":"Plasmodium","strain":""}]},{"year":2022,"claim":"The cellular role of the parasite complex was unknown; reciprocal Co-IP defined a TgATG12–TgATG5–TgATG16L complex that localizes to the apicoplast periphery, and inducible depletion of TgATG12 caused apicoplast loss and growth impairment, establishing an essential role in apicoplast biogenesis.","evidence":"Co-immunoprecipitation, inducible auxin-degron knockdown, fluorescence microscopy, and parasite growth assay","pmids":["36095096"],"confidence":"High","gaps":["Mechanism linking ATG8 conjugation to apicoplast maintenance not fully defined","Strain background not specified"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2022,"claim":"How the complex directs TgATG8 to the apicoplast was unknown; mutation of a ubiquitin-interacting-motif-docking site on TgATG8 abolished apicoplast localization without affecting lipidation, and TgSNAP29 was required for vesicular translocation, separating conjugation from targeting.","evidence":"Site-directed UDS mutagenesis, inducible knockdown, fluorescence microscopy, and lipidation assay","pmids":["36095096"],"confidence":"High","gaps":["Identity of the UDS-interacting partner not defined","How SNAP29-dependent vesicles reach the apicoplast unresolved"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":null,"claim":"The receptor or signal that positions the ATG12–ATG5–ATG16L(1) complex on its correct native membrane — the apicoplast in the parasite and the PVM in the host — remains undefined.","evidence":"","pmids":[],"confidence":"High","gaps":["No membrane-recruitment receptor identified in either system","Structural model of the parasite non-covalent complex on membranes lacking"],"attribution":[]}],"mechanism_profile":{"molecular_activity":[],"localization":[],"pathway":[],"complexes":["TgATG12–TgATG5–TgATG16L complex (non-covalent, apicoplast periphery)","Host ATG12–ATG5–ATG16L1 complex (with ATG7 and ATG3)"],"partners":["ATG5","ATG16L / ATG16L1","ATG8 / LC3, GABARAP, GABARAPL1, GABARAPL2","TGSNAP29","ATG7","ATG3"],"other_free_text":["TgATG12 lacks the canonical C-terminal glycine and its E2-like enzyme ATG10 is absent, forming a non-covalent rather than covalent complex with ATG5 [#0].","A ubiquitin-interacting-motif-docking site (UDS) on TgATG8 is required for apicoplast localization independent of lipidation [#2].","Relocating the host complex to alternative membranes is sufficient to redirect LC3 homologs and IFN-γ-inducible GTPases [#4]."]},"strain_conflicts":[]},"prefetch_data":{},"corpus_meta":[{"pmid":"24931121","id":"PMC_24931121","title":"The parasitophorous vacuole membrane of Toxoplasma gondii is targeted for disruption by ubiquitin-like conjugation systems of autophagy.","date":"2014","source":"Immunity","url":"https://pubmed.ncbi.nlm.nih.gov/24931121","citation_count":159,"is_preprint":false},{"pmid":"27172324","id":"PMC_27172324","title":"Targeting by AutophaGy proteins (TAG): Targeting of IFNG-inducible GTPases to membranes by the LC3 conjugation system of autophagy.","date":"2016","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/27172324","citation_count":72,"is_preprint":false},{"pmid":"30911187","id":"PMC_30911187","title":"Evolution from covalent conjugation to non-covalent interaction in the ubiquitin-like ATG12 system.","date":"2019","source":"Nature structural & molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/30911187","citation_count":45,"is_preprint":false},{"pmid":"36095096","id":"PMC_36095096","title":"Apicoplast biogenesis mediated by ATG8 requires the ATG12-ATG5-ATG16L and SNAP29 complexes in Toxoplasma gondii.","date":"2022","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/36095096","citation_count":24,"is_preprint":false},{"pmid":"35757711","id":"PMC_35757711","title":"Toxoplasma gondii CDPK3 Controls the Intracellular Proliferation of Parasites in Macrophages.","date":"2022","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/35757711","citation_count":8,"is_preprint":false},{"pmid":"34453672","id":"PMC_34453672","title":"Toxoplasma gondii profilin and tachyzoites RH strain may manipulate autophagy via downregulating Atg5 and Atg12 and upregulating Atg7.","date":"2021","source":"Molecular biology reports","url":"https://pubmed.ncbi.nlm.nih.gov/34453672","citation_count":7,"is_preprint":false},{"pmid":"36941573","id":"PMC_36941573","title":"Evaluation of the mTORC activity in the presence of Toxoplasma gondii and azathioprine in human monocyte cell line.","date":"2023","source":"BMC microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/36941573","citation_count":1,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":6550,"output_tokens":1768,"usd":0.023085,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":6890,"output_tokens":3100,"usd":0.055975,"stage2_stop_reason":"end_turn"},"total_usd":0.07906,"stage1_batch_id":"msgbatch_01S6GjoYQm8LTPUsdhASdqag","stage2_batch_id":"msgbatch_01VX63RTMdBJqsG6gQMqoCjz","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2019,\n      \"finding\": \"In Toxoplasma gondii (and Plasmodium), ATG12 lacks the C-terminal glycine required for covalent conjugation and the E2-like enzyme ATG10 is absent; instead, TgATG12 forms a non-covalent complex with ATG5. This non-covalent ATG12-ATG5 complex retains the ability to facilitate ATG8-phosphatidylethanolamine conjugation, demonstrating an evolutionary transition from covalent conjugation to non-covalent interaction in the ATG12 system.\",\n      \"method\": \"Biochemical reconstitution, sequence/structural analysis, in vitro ATG8 lipidation assay\",\n      \"journal\": \"Nature structural & molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution of non-covalent complex with functional ATG8 lipidation assay, supported by sequence analysis across multiple organisms\",\n      \"pmids\": [\"30911187\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Toxoplasma gondii\", \"strain\": \"\"},\n        {\"species\": \"Plasmodium\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"TgATG12 forms a complex with TgATG5 and TgATG16L in Toxoplasma gondii. This TgATG12-TgATG5-TgATG16L complex localizes to isolated structures at the periphery of the apicoplast in a TgATG16L-dependent manner. Inducible depletion of TgATG12 caused loss of the apicoplast and impaired parasite growth, establishing a required role for TgATG12 in apicoplast biogenesis.\",\n      \"method\": \"Co-immunoprecipitation, inducible auxin-degron knockdown, fluorescence microscopy, parasite growth assay\",\n      \"journal\": \"Autophagy\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP, inducible depletion with specific apicoplast phenotype, multiple orthogonal methods in one study\",\n      \"pmids\": [\"36095096\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"The TgATG12-TgATG5-TgATG16L complex is required for TgATG8 localization to the apicoplast. A putative ubiquitin-interacting motif-docking site (UDS) on TgATG8 was identified; mutation of this UDS abolished TgATG8 localization on the apicoplast without affecting ATG8 lipidation, and a SNARE protein TgSNAP29 was required for TgATG8 vesicular translocation to the apicoplast.\",\n      \"method\": \"Site-directed mutagenesis of UDS, inducible knockdown (AID), fluorescence microscopy, lipidation assay\",\n      \"journal\": \"Autophagy\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — mutagenesis of functional site combined with inducible depletion and multiple readouts in one study\",\n      \"pmids\": [\"36095096\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"The host ATG12-ATG5-ATG16L1 complex (together with ATG7 and ATG3) is required for targeting LC3 and IFN-γ effectors to the parasitophorous vacuole membrane (PVM) of T. gondii and for subsequent disruption of the PVM. Pharmacological modulation of degradative autophagy or deletion of other essential autophagy genes had no substantial effect, indicating specificity of the LC3 conjugation system in this immune function.\",\n      \"method\": \"Genetic knockout of host autophagy genes, in vitro infection assay, in vivo mouse model, immunofluorescence microscopy\",\n      \"journal\": \"Immunity\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean genetic knockouts in multiple autophagy components with specific mechanistic readout (LC3/effector targeting to PVM), replicated in vitro and in vivo\",\n      \"pmids\": [\"24931121\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Mus musculus\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"The host ATG12-ATG5-ATG16L1 complex specifies the conjugation site of LC3 homologs (LC3, GABARAP, GABARAPL1, GABARAPL2) on the PVM of T. gondii. Experimental relocation of the ATG12-ATG5-ATG16L1 complex to alternative membranes redirected IFN-γ-inducible GTPase targeting to those new membranes rather than the PVM, demonstrating that ATG12-ATG5-ATG16L1 complex localization is necessary and sufficient to determine the membrane destination of LC3 homologs and downstream immune effectors.\",\n      \"method\": \"Knockout/knockdown of LC3 homologs, artificial membrane targeting of ATG12-ATG5-ATG16L1 complex, immunofluorescence, infection assay\",\n      \"journal\": \"Autophagy\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — gain-of-function membrane retargeting experiment combined with loss-of-function genetics, multiple orthogonal methods, mechanistically definitive\",\n      \"pmids\": [\"27172324\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Mus musculus\", \"strain\": \"\"}\n      ]\n    }\n  ],\n  \"current_model\": \"In Toxoplasma gondii, ATG12 lacks the canonical C-terminal glycine and the E2-like enzyme ATG10, so it forms a non-covalent (rather than covalent) complex with ATG5; this TgATG12–TgATG5–TgATG16L complex localizes to the apicoplast periphery, is essential for apicoplast biogenesis and parasite viability, and facilitates TgATG8–phosphatidylethanolamine conjugation; in the host cell, the orthologous ATG12–ATG5–ATG16L1 complex specifies where LC3 homologs are conjugated to membranes and thereby directs IFN-γ immune effectors to the parasitophorous vacuole membrane for its disruption.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"ATG12 is a core component of the ATG12–ATG5–ATG16L conjugation machinery that specifies where ATG8/LC3-family proteins are conjugated to membranes, a function conserved from apicomplexan parasites to their mammalian hosts [#0, #1, #3]. In Toxoplasma gondii, ATG12 has diverged from the canonical ubiquitin-like pathway: it lacks the C-terminal glycine required for covalent conjugation and its cognate E2-like enzyme ATG10 is absent, so TgATG12 instead forms a non-covalent complex with ATG5 that nonetheless retains the ability to promote ATG8–phosphatidylethanolamine conjugation [#0]. This TgATG12–TgATG5–TgATG16L complex localizes to the apicoplast periphery in a TgATG16L-dependent manner and is required for apicoplast biogenesis and parasite growth [#1], directing TgATG8 to the apicoplast via a ubiquitin-interacting-motif-docking site on TgATG8 and SNAP29-dependent vesicular translocation [#2]. In the mammalian host, the orthologous ATG12–ATG5–ATG16L1 complex is required to target LC3 homologs and IFN-γ effectors to the T. gondii parasitophorous vacuole membrane (PVM) and disrupt it [#3], and the complex's own membrane localization is both necessary and sufficient to determine where LC3 homologs and downstream IFN-γ-inducible GTPases are deposited [#4].\",\n  \"teleology\": [\n    {\n      \"year\": 2014,\n      \"claim\": \"It was unknown which autophagy machinery directs anti-Toxoplasma IFN-γ immunity; genetic dissection showed the LC3 conjugation system, not degradative autophagy, targets LC3 and IFN-γ effectors to the PVM for its disruption, establishing a non-canonical immune role for ATG12–ATG5–ATG16L1.\",\n      \"evidence\": \"Genetic knockout of host autophagy genes with in vitro infection and in vivo mouse models plus immunofluorescence\",\n      \"pmids\": [\"24931121\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not resolve how the complex is recruited specifically to the PVM\", \"Did not distinguish the roles of individual complex members\"],\n      \"attribution\": [{\"species\": \"Mus musculus\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Whether ATG12–ATG5–ATG16L1 merely conjugates LC3 or dictates its destination was unresolved; retargeting the complex to alternative membranes redirected LC3 homologs and IFN-γ GTPases there, establishing that complex localization is necessary and sufficient to specify the conjugation site.\",\n      \"evidence\": \"Loss-of-function knockdown of LC3 homologs combined with gain-of-function artificial membrane targeting, immunofluorescence, and infection assays\",\n      \"pmids\": [\"27172324\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not identify the receptor that positions the complex on native target membranes\", \"Restricted to the host anti-parasite context\"],\n      \"attribution\": [{\"species\": \"Mus musculus\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"It was unclear whether ATG12 function requires covalent conjugation in all lineages; reconstitution showed T. gondii and Plasmodium ATG12 lack the C-terminal glycine and ATG10, forming a non-covalent ATG12–ATG5 complex that still supports ATG8 lipidation, revealing an evolutionary shift from covalent to non-covalent assembly.\",\n      \"evidence\": \"Biochemical reconstitution, sequence/structural analysis, and in vitro ATG8 lipidation assay\",\n      \"pmids\": [\"30911187\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not establish the in-cell consequences of the non-covalent complex\", \"Structural basis of the non-covalent interaction not resolved\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}, {\"species\": \"Plasmodium\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"The cellular role of the parasite complex was unknown; reciprocal Co-IP defined a TgATG12–TgATG5–TgATG16L complex that localizes to the apicoplast periphery, and inducible depletion of TgATG12 caused apicoplast loss and growth impairment, establishing an essential role in apicoplast biogenesis.\",\n      \"evidence\": \"Co-immunoprecipitation, inducible auxin-degron knockdown, fluorescence microscopy, and parasite growth assay\",\n      \"pmids\": [\"36095096\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism linking ATG8 conjugation to apicoplast maintenance not fully defined\", \"Strain background not specified\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"How the complex directs TgATG8 to the apicoplast was unknown; mutation of a ubiquitin-interacting-motif-docking site on TgATG8 abolished apicoplast localization without affecting lipidation, and TgSNAP29 was required for vesicular translocation, separating conjugation from targeting.\",\n      \"evidence\": \"Site-directed UDS mutagenesis, inducible knockdown, fluorescence microscopy, and lipidation assay\",\n      \"pmids\": [\"36095096\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of the UDS-interacting partner not defined\", \"How SNAP29-dependent vesicles reach the apicoplast unresolved\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The receptor or signal that positions the ATG12–ATG5–ATG16L(1) complex on its correct native membrane — the apicoplast in the parasite and the PVM in the host — remains undefined.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No membrane-recruitment receptor identified in either system\", \"Structural model of the parasite non-covalent complex on membranes lacking\"],\n      \"attribution\": []\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0019776\", \"supporting_discovery_ids\": [0], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}, {\"species\": \"Plasmodium\", \"strain\": \"\"}]},\n      {\"term_id\": \"GO:0061919\", \"supporting_discovery_ids\": [0, 2, 3, 4], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}, {\"species\": \"Mus musculus\", \"strain\": \"\"}]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0020011\", \"supporting_discovery_ids\": [1, 2], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]},\n      {\"term_id\": \"GO:0097470\", \"supporting_discovery_ids\": [3, 4], \"attribution\": [{\"species\": \"Mus musculus\", \"strain\": \"\"}]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0000045\", \"supporting_discovery_ids\": [0, 1, 2], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]},\n      {\"term_id\": \"GO:0071222\", \"supporting_discovery_ids\": [3, 4], \"attribution\": [{\"species\": \"Mus musculus\", \"strain\": \"\"}]}\n    ],\n    \"complexes\": [\n      \"TgATG12–TgATG5–TgATG16L complex (non-covalent, apicoplast periphery)\",\n      \"Host ATG12–ATG5–ATG16L1 complex (with ATG7 and ATG3)\"\n    ],\n    \"partners\": [\n      \"ATG5\",\n      \"ATG16L / ATG16L1\",\n      \"ATG8 / LC3, GABARAP, GABARAPL1, GABARAPL2\",\n      \"TgSNAP29\",\n      \"ATG7\",\n      \"ATG3\"\n    ],\n    \"other_free_text\": [\n      \"TgATG12 lacks the canonical C-terminal glycine and its E2-like enzyme ATG10 is absent, forming a non-covalent rather than covalent complex with ATG5 [#0].\",\n      \"A ubiquitin-interacting-motif-docking site (UDS) on TgATG8 is required for apicoplast localization independent of lipidation [#2].\",\n      \"Relocating the host complex to alternative membranes is sufficient to redirect LC3 homologs and IFN-γ-inducible GTPases [#4].\"\n    ]\n  },\n  \"strain_conflicts\": []\n}","audit_flag":null,"strains":[{"strain":"ME49","gene_id":"TGME49_321300","name":"ATG12","product":"autophagy-related protein ATG12","uniprot_acc":"S8GK31","chromosome":"TGME49_chrIb","start_pos":1894311},{"strain":"GT1","gene_id":"TGGT1_321300","name":null,"product":"putative autophagy-related 12 variant 1","uniprot_acc":null,"chromosome":"TGGT1_chrIb","start_pos":1902556},{"strain":"RH88","gene_id":"TGRH88_020760","name":null,"product":"putative autophagy-related 12 variant 1","uniprot_acc":null,"chromosome":"CM023082","start_pos":68181},{"strain":"VEG","gene_id":"TGVEG_321300","name":null,"product":"putative autophagy-related 12 variant 1","uniprot_acc":"V5B115","chromosome":"TGVEG_chrIb","start_pos":1912838}]}