{"gene":"TG_289680","gene_symbol":"RAB11A","run_date":"2026-09-08T14:08:30","timeline":{"discoveries":[{"year":2009,"finding":"Rab11A is required for delivery of new plasma membrane to daughter cells during apicomplexan cytokinesis; ablation of Rab11A function results in daughters with incompletely formed Inner Membrane Complex (IMC) and a block at a late stage of cell division.","method":"Dominant-negative/ablation of Rab11A function with phenotypic readout (incomplete IMC, block in cytokinesis); live imaging","journal":"PLoS pathogens","confidence":"High","confidence_rationale":"Tier 2 / Strong — loss-of-function with defined cellular phenotype (incomplete IMC, cytokinesis block), complemented by epistasis with myosin A tail mutant; published in peer-reviewed journal with clear mechanistic model","pmids":["19165333"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""},{"species":"Plasmodium falciparum","strain":""}]},{"year":2009,"finding":"Rab11A physically associates with Myosin-Tail-Interacting-Protein (MTIP/MLC1), a component of the glideosome motor complex, suggesting Rab11A-mediated vesicular traffic is driven by an MTIP-Myosin motor for IMC maturation.","method":"Co-immunoprecipitation / association assay between Rab11A and MTIP","journal":"PLoS pathogens","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — co-association demonstrated in a single study; single lab but supported by parallel genetic epistasis with myosin A tail mutant","pmids":["19165333"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""},{"species":"Plasmodium falciparum","strain":""}]},{"year":2017,"finding":"Host Rab11A-positive vesicles are sequestered into the Toxoplasma parasitophorous vacuole (PV) and concentrate at the intravacuolar network (IVN); this accumulation is reduced in Δgra2 and Δgra2Δgra6 parasites, implicating TgGRA2 and TgGRA6 in stabilizing tubules that facilitate host vesicle sequestration.","method":"Correlative light and electron microscopy; genetic deletion of gra2 and gra6 with quantitative microscopic readout of intra-PV Rab11A vesicles","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis (Δgra2, Δgra2Δgra6) with quantitative microscopic phenotype, plus correlative EM; peer-reviewed, multiple orthogonal methods","pmids":["29070609"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2017,"finding":"Overexpression of the phospholipase TgLCAT, localized to the IVN, decreases the number of intravacuolar GFP-Rab11A vesicles, indicating that TgLCAT controls lipolytic degradation of sequestered host Rab11A vesicles for cargo release.","method":"Overexpression of TgLCAT followed by quantitative microscopy of intra-PV GFP-Rab11A vesicles","journal":"The Journal of cell biology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — single overexpression experiment with quantitative readout; single lab, one method","pmids":["29070609"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2020,"finding":"Rab11A promotes cytoskeleton-driven transport of dense granules and their constitutive secretion into the vacuolar space; Rab11A-defective parasites show impaired dense granule transport and reduced secretion of dense granule cargo.","method":"Loss-of-function of Rab11A (dominant-negative/conditional), live imaging, secretion assays","journal":"PLoS pathogens","confidence":"High","confidence_rationale":"Tier 2 / Moderate — loss-of-function with live imaging of dense granule transport and secretion assays, multiple readouts in a single peer-reviewed study","pmids":["32463830"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2020,"finding":"Rab11A regulates transmembrane protein trafficking and localization at the plasma membrane during parasite replication, indicating a broad role in cargo exocytosis.","method":"Loss-of-function of Rab11A with imaging of transmembrane protein localization","journal":"PLoS pathogens","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — single lab, imaging-based localization with loss-of-function, single study","pmids":["32463830"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2020,"finding":"Rab11A-positive vesicles and dense granules accumulate in a polarized fashion at the apical pole of extracellular motile and invading parasites, and MIC2 secretion is altered in Rab11A-defective parasites, indicating that apically polarized Rab11A-dependent delivery regulates early secretory events during host cell invasion.","method":"Live imaging of Rab11A-GFP in motile/invading parasites; MIC2 secretion assay in Rab11A-defective parasites","journal":"PLoS pathogens","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — live imaging with functional readout (MIC2 secretion), loss-of-function in a single peer-reviewed study with two orthogonal methods","pmids":["32463830"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2021,"finding":"TgCDPK7 phosphorylates TgRab11A, and this phosphorylation is critical for parasite intracellular development and protein trafficking; identified by quantitative phosphoproteomics of TgCDPK7-knockout parasites.","method":"Quantitative phosphoproteomics of TgCDPK7 knockout; functional validation of phosphorylation site importance for intracellular development and protein trafficking","journal":"PLoS pathogens","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — phosphoproteomics combined with functional validation of phosphorylation in a single peer-reviewed study; single lab","pmids":["33635921"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2022,"finding":"Host Rab11A vesicle recruitment into the Toxoplasma PV is selective: depletion of host Rab11A reduces intra-PV FIP5 (class I FIP)-positive vesicles but not FIP3 (class II FIP)-positive vesicles, demonstrating that host Rab11A specifically mediates class I FIP vesicle delivery to the PV.","method":"Cell depletion of Rab11A combined with quantitative microscopy of intra-PV FIP-positive vesicles; dominant-negative FIP3 mutants abolishing Rab11 and Arf6 binding","journal":"Molecular biology of the cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — cell depletion with quantitative microscopy and mutant rescue, single lab, single study","pmids":["35274991"],"is_preprint":false,"attribution":[{"species":"Homo sapiens","strain":""}]},{"year":2024,"finding":"TBC9, a TBC-domain Rab GAP localized to the ER, physically interacts with Rab11A (and Rab11B and Rab2) as shown by pull-down assay, supporting a role for TBC9 as a GAP regulator of Rab11A in early vesicular transport and daughter IMC formation.","method":"Pull-down assay between TBC9 and Rab GTPases (Rab11A, Rab11B, Rab2); localization by fluorescence imaging; TBC9 loss-of-function phenotype analysis","journal":"Communications biology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — single pull-down showing TBC9–Rab11A interaction, supported by loss-of-function phenotype; single lab, one method for the interaction","pmids":["38762629"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2022,"finding":"Host Rab11A abundance decreases in atg5-/- HeLa cells infected with Toxoplasma (compared to stable levels in wild-type cells), and disruption of host rab11a in HeLa cells compromises T. gondii proliferation and increases gra2 transcription in the parasite, indicating that host autophagy limits parasite access to Rab11A-positive vesicles to restrict parasite replication.","method":"CRISPR/Cas9 knockout of host atg5 and rab11a; qRT-PCR; intracellular replication assays","journal":"Frontiers in microbiology","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — CRISPR/Cas9 knockouts with replication readout, but purely focused on host Rab11A (not parasite Rab11A), single lab, single study","pmids":["36532461"],"is_preprint":false,"attribution":[{"species":"Homo sapiens","strain":"HeLa"},{"species":"Toxoplasma gondii","strain":"RH"}]}],"current_model":"Toxoplasma gondii Rab11A is a small GTPase that controls vesicular trafficking at multiple stages of the parasite life cycle: it mediates delivery of new plasma membrane and IMC components to daughter cells during cytokinesis (interacting with the MTIP/MLC1–MyosinA motor complex), drives cytoskeleton-dependent constitutive transport and apically polarized secretion of dense granules (regulating MIC2 release and host cell invasion), and is phosphorylated by TgCDPK7 to regulate intracellular development and protein trafficking; on the host side, Toxoplasma selectively sequesters host Rab11A-positive vesicles into the parasitophorous vacuole intravacuolar network in a TgGRA2/GRA6-dependent manner, where TgLCAT mediates their lipolytic processing for nutrient acquisition."},"narrative":{"mechanistic_narrative":"Toxoplasma gondii Rab11A is a small GTPase that governs vesicular trafficking at multiple stages of the parasite life cycle by directing the delivery of membrane and secretory cargo to specific cellular destinations [PMID:19165333, PMID:32463830]. During cytokinesis it supplies new plasma membrane to nascent daughter cells, and its loss produces daughters with incompletely formed Inner Membrane Complex and a block at a late stage of division; this membrane traffic is coupled to the glideosome motor through physical association with MTIP/MLC1, implicating an MTIP-Myosin motor in Rab11A-driven IMC maturation [PMID:19165333]. Rab11A additionally drives cytoskeleton-dependent transport and constitutive secretion of dense granules, regulates transmembrane protein delivery to the plasma membrane during replication, and mediates apically polarized delivery that controls MIC2 secretion and early secretory events during host cell invasion [PMID:32463830]. Its activity is regulated by phosphorylation by TgCDPK7, which is critical for intracellular development and protein trafficking, and by the ER-localized TBC-domain GAP TBC9, which physically interacts with Rab11A and functions in early vesicular transport and daughter IMC formation [PMID:33635921, PMID:38762629]. On the host side, Toxoplasma sequesters host Rab11A-positive class I FIP vesicles into the parasitophorous vacuole intravacuolar network in a TgGRA2/GRA6-dependent manner, where they are lipolytically processed by the phospholipase TgLCAT, and host autophagy limits parasite access to these vesicles to restrict replication [PMID:29070609, PMID:35274991, PMID:36532461].","teleology":[{"year":2009,"claim":"Established the first cellular role for apicomplexan Rab11A by asking what drives membrane delivery during parasite cell division, showing it is required to build daughter cell surfaces.","evidence":"Dominant-negative/ablation of Rab11A with live imaging and phenotypic readout of IMC formation and cytokinesis, plus association with the MTIP/MLC1 glideosome component","pmids":["19165333"],"confidence":"High","gaps":["Direct GTPase cycle/effector biochemistry not resolved","MTIP association shown by a single co-IP without reciprocal validation","Vesicle cargo composition not defined"],"attribution":[{"species":"Toxoplasma gondii","strain":""},{"species":"Plasmodium falciparum","strain":""}]},{"year":2017,"claim":"Extended Rab11A biology to the host-parasite interface by asking whether Toxoplasma exploits host Rab11A vesicles, demonstrating selective sequestration into the PV and their lipolytic processing.","evidence":"Correlative light/EM plus genetic deletion of gra2/gra6 and TgLCAT overexpression with quantitative microscopy of intra-PV GFP-Rab11A vesicles","pmids":["29070609"],"confidence":"High","gaps":["Mechanism by which GRA2/GRA6 stabilize tubules for capture not resolved","TgLCAT-Rab11A degradation link rests on single overexpression experiment","Nutrient/cargo actually acquired not directly identified"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2020,"claim":"Defined the secretory scope of parasite Rab11A by asking which trafficking events it controls, revealing roles in dense granule transport/secretion, transmembrane protein delivery, and apically polarized MIC2 secretion during invasion.","evidence":"Loss-of-function of Rab11A with live imaging of dense granule and Rab11A-GFP dynamics plus secretion assays including MIC2","pmids":["32463830"],"confidence":"High","gaps":["Effectors linking Rab11A to the cytoskeleton not identified","How apical polarization of Rab11A vesicles is established is unknown","Direct versus indirect effect on MIC2 secretion not dissected"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2021,"claim":"Addressed how parasite Rab11A activity is regulated by identifying it as a substrate of TgCDPK7, linking a kinase signal to trafficking and intracellular development.","evidence":"Quantitative phosphoproteomics of TgCDPK7 knockout with functional validation of phosphosite importance","pmids":["33635921"],"confidence":"Medium","gaps":["Functional consequence of the specific phosphorylation on Rab11A GTPase activity not defined","Whether phosphorylation alters effector binding unknown","Single-study phosphoproteomic identification"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2022,"claim":"Refined the host vesicle recruitment mechanism by asking which host Rab11A vesicle subclass is captured, showing selectivity for class I FIP (FIP5) but not class II FIP (FIP3) vesicles, and demonstrating host autophagy restricts parasite access.","evidence":"Host Rab11A depletion with quantitative microscopy of intra-PV FIP vesicles and dominant-negative FIP3 mutants (MBoC); CRISPR knockout of host atg5/rab11a with replication assays and qRT-PCR (Front Microbiol)","pmids":["35274991","36532461"],"confidence":"Medium","gaps":["Parasite receptor/factor that discriminates FIP subclasses not identified","Mechanistic link between autophagy and Rab11A vesicle availability unresolved","Studies focus on host Rab11A, not parasite Rab11A"],"attribution":[{"species":"Homo sapiens","strain":""},{"species":"Homo sapiens","strain":"HeLa"},{"species":"Toxoplasma gondii","strain":"RH"}]},{"year":2024,"claim":"Identified a candidate regulatory GAP by asking what controls the parasite Rab11A GTPase cycle, showing the ER-localized TBC9 interacts with Rab11A and functions in early transport and daughter IMC formation.","evidence":"Pull-down assay of TBC9 with Rab11A/Rab11B/Rab2, fluorescence localization, and TBC9 loss-of-function phenotyping","pmids":["38762629"],"confidence":"Medium","gaps":["GAP activity on Rab11A not biochemically demonstrated","Interaction shown by a single pull-down without reciprocal validation","TBC9 binds multiple Rabs, so Rab11A specificity of its function is unclear"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":null,"claim":"How Rab11A GTPase cycling, phosphorylation, and GAP regulation are integrated with its distinct effector complexes at each trafficking step remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural or biochemical map of Rab11A GEF/GAP/effector cycle in the parasite","Direct effectors linking Rab11A to cytoskeletal transport not identified","Consequences of TgCDPK7 phosphorylation on nucleotide state and effector engagement unknown"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]}],"mechanism_profile":{"molecular_activity":[],"localization":[],"pathway":[],"complexes":["MTIP/MLC1-MyosinA glideosome motor association"],"partners":["MTIP/MLC1 (GLIDEOSOME, T. GONDII)","TGCDPK7 (KINASE, T. GONDII)","TBC9 (ER-LOCALIZED TBC-DOMAIN RAB GAP, T. GONDII)","TGGRA2 / TGGRA6 (HOST VESICLE SEQUESTRATION, T. GONDII)","TGLCAT (PHOSPHOLIPASE, T. GONDII)","FIP5 / CLASS I FIP VESICLES (HOST)"],"other_free_text":["Host Rab11A-positive class I FIP (FIP5) vesicles are selectively recruited to the PV; class II FIP (FIP3) vesicles are not.","Host autophagy (ATG5-dependent) limits parasite access to Rab11A vesicles and restricts replication; rab11a disruption increases parasite gra2 transcription.","TBC9 interacts with Rab11A, Rab11B, and Rab2, so its regulatory function is not exclusive to Rab11A."]},"strain_conflicts":[]},"prefetch_data":{},"corpus_meta":[{"pmid":"19165333","id":"PMC_19165333","title":"Rab11A-controlled assembly of the inner membrane complex is required for completion of apicomplexan cytokinesis.","date":"2009","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/19165333","citation_count":95,"is_preprint":false},{"pmid":"29070609","id":"PMC_29070609","title":"The parasite Toxoplasma sequesters diverse Rab host vesicles within an intravacuolar network.","date":"2017","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/29070609","citation_count":73,"is_preprint":false},{"pmid":"32463830","id":"PMC_32463830","title":"Rab11A regulates dense granule transport and secretion during Toxoplasma gondii invasion of host cells and parasite replication.","date":"2020","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/32463830","citation_count":32,"is_preprint":false},{"pmid":"33635921","id":"PMC_33635921","title":"Protein kinase TgCDPK7 regulates vesicular trafficking and phospholipid synthesis in Toxoplasma gondii.","date":"2021","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/33635921","citation_count":20,"is_preprint":false},{"pmid":"35274991","id":"PMC_35274991","title":"The Rab11-family interacting proteins reveal selective interaction of mammalian recycling endosomes with the Toxoplasma parasitophorous vacuole in a Rab11- and Arf6-dependent manner.","date":"2022","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/35274991","citation_count":8,"is_preprint":false},{"pmid":"38762629","id":"PMC_38762629","title":"TBC9, an essential TBC-domain protein, regulates early vesicular transport and IMC formation in Toxoplasma gondii.","date":"2024","source":"Communications biology","url":"https://pubmed.ncbi.nlm.nih.gov/38762629","citation_count":5,"is_preprint":false},{"pmid":"36532461","id":"PMC_36532461","title":"Host autophagy limits Toxoplasma gondii proliferation in the absence of IFN-γ by affecting the hijack of Rab11A-positive vesicles.","date":"2022","source":"Frontiers in microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/36532461","citation_count":3,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":5855,"output_tokens":3136,"usd":0.032302,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":8815,"output_tokens":4004,"usd":0.072087,"stage2_stop_reason":"end_turn"},"total_usd":0.104389,"stage1_batch_id":"msgbatch_015xmQKDRqnTh9anE4m7LiD6","stage2_batch_id":"msgbatch_01JLpkFTVhoQBNgqHCKFrVVU","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2009,\n      \"finding\": \"Rab11A is required for delivery of new plasma membrane to daughter cells during apicomplexan cytokinesis; ablation of Rab11A function results in daughters with incompletely formed Inner Membrane Complex (IMC) and a block at a late stage of cell division.\",\n      \"method\": \"Dominant-negative/ablation of Rab11A function with phenotypic readout (incomplete IMC, block in cytokinesis); live imaging\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — loss-of-function with defined cellular phenotype (incomplete IMC, cytokinesis block), complemented by epistasis with myosin A tail mutant; published in peer-reviewed journal with clear mechanistic model\",\n      \"pmids\": [\"19165333\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Toxoplasma gondii\", \"strain\": \"\"},\n        {\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Rab11A physically associates with Myosin-Tail-Interacting-Protein (MTIP/MLC1), a component of the glideosome motor complex, suggesting Rab11A-mediated vesicular traffic is driven by an MTIP-Myosin motor for IMC maturation.\",\n      \"method\": \"Co-immunoprecipitation / association assay between Rab11A and MTIP\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — co-association demonstrated in a single study; single lab but supported by parallel genetic epistasis with myosin A tail mutant\",\n      \"pmids\": [\"19165333\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Toxoplasma gondii\", \"strain\": \"\"},\n        {\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Host Rab11A-positive vesicles are sequestered into the Toxoplasma parasitophorous vacuole (PV) and concentrate at the intravacuolar network (IVN); this accumulation is reduced in Δgra2 and Δgra2Δgra6 parasites, implicating TgGRA2 and TgGRA6 in stabilizing tubules that facilitate host vesicle sequestration.\",\n      \"method\": \"Correlative light and electron microscopy; genetic deletion of gra2 and gra6 with quantitative microscopic readout of intra-PV Rab11A vesicles\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis (Δgra2, Δgra2Δgra6) with quantitative microscopic phenotype, plus correlative EM; peer-reviewed, multiple orthogonal methods\",\n      \"pmids\": [\"29070609\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Overexpression of the phospholipase TgLCAT, localized to the IVN, decreases the number of intravacuolar GFP-Rab11A vesicles, indicating that TgLCAT controls lipolytic degradation of sequestered host Rab11A vesicles for cargo release.\",\n      \"method\": \"Overexpression of TgLCAT followed by quantitative microscopy of intra-PV GFP-Rab11A vesicles\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — single overexpression experiment with quantitative readout; single lab, one method\",\n      \"pmids\": [\"29070609\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Rab11A promotes cytoskeleton-driven transport of dense granules and their constitutive secretion into the vacuolar space; Rab11A-defective parasites show impaired dense granule transport and reduced secretion of dense granule cargo.\",\n      \"method\": \"Loss-of-function of Rab11A (dominant-negative/conditional), live imaging, secretion assays\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with live imaging of dense granule transport and secretion assays, multiple readouts in a single peer-reviewed study\",\n      \"pmids\": [\"32463830\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Rab11A regulates transmembrane protein trafficking and localization at the plasma membrane during parasite replication, indicating a broad role in cargo exocytosis.\",\n      \"method\": \"Loss-of-function of Rab11A with imaging of transmembrane protein localization\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — single lab, imaging-based localization with loss-of-function, single study\",\n      \"pmids\": [\"32463830\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Rab11A-positive vesicles and dense granules accumulate in a polarized fashion at the apical pole of extracellular motile and invading parasites, and MIC2 secretion is altered in Rab11A-defective parasites, indicating that apically polarized Rab11A-dependent delivery regulates early secretory events during host cell invasion.\",\n      \"method\": \"Live imaging of Rab11A-GFP in motile/invading parasites; MIC2 secretion assay in Rab11A-defective parasites\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — live imaging with functional readout (MIC2 secretion), loss-of-function in a single peer-reviewed study with two orthogonal methods\",\n      \"pmids\": [\"32463830\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"TgCDPK7 phosphorylates TgRab11A, and this phosphorylation is critical for parasite intracellular development and protein trafficking; identified by quantitative phosphoproteomics of TgCDPK7-knockout parasites.\",\n      \"method\": \"Quantitative phosphoproteomics of TgCDPK7 knockout; functional validation of phosphorylation site importance for intracellular development and protein trafficking\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — phosphoproteomics combined with functional validation of phosphorylation in a single peer-reviewed study; single lab\",\n      \"pmids\": [\"33635921\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Host Rab11A vesicle recruitment into the Toxoplasma PV is selective: depletion of host Rab11A reduces intra-PV FIP5 (class I FIP)-positive vesicles but not FIP3 (class II FIP)-positive vesicles, demonstrating that host Rab11A specifically mediates class I FIP vesicle delivery to the PV.\",\n      \"method\": \"Cell depletion of Rab11A combined with quantitative microscopy of intra-PV FIP-positive vesicles; dominant-negative FIP3 mutants abolishing Rab11 and Arf6 binding\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — cell depletion with quantitative microscopy and mutant rescue, single lab, single study\",\n      \"pmids\": [\"35274991\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Homo sapiens\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"TBC9, a TBC-domain Rab GAP localized to the ER, physically interacts with Rab11A (and Rab11B and Rab2) as shown by pull-down assay, supporting a role for TBC9 as a GAP regulator of Rab11A in early vesicular transport and daughter IMC formation.\",\n      \"method\": \"Pull-down assay between TBC9 and Rab GTPases (Rab11A, Rab11B, Rab2); localization by fluorescence imaging; TBC9 loss-of-function phenotype analysis\",\n      \"journal\": \"Communications biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — single pull-down showing TBC9–Rab11A interaction, supported by loss-of-function phenotype; single lab, one method for the interaction\",\n      \"pmids\": [\"38762629\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Host Rab11A abundance decreases in atg5-/- HeLa cells infected with Toxoplasma (compared to stable levels in wild-type cells), and disruption of host rab11a in HeLa cells compromises T. gondii proliferation and increases gra2 transcription in the parasite, indicating that host autophagy limits parasite access to Rab11A-positive vesicles to restrict parasite replication.\",\n      \"method\": \"CRISPR/Cas9 knockout of host atg5 and rab11a; qRT-PCR; intracellular replication assays\",\n      \"journal\": \"Frontiers in microbiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — CRISPR/Cas9 knockouts with replication readout, but purely focused on host Rab11A (not parasite Rab11A), single lab, single study\",\n      \"pmids\": [\"36532461\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Homo sapiens\", \"strain\": \"HeLa\"},\n        {\"species\": \"Toxoplasma gondii\", \"strain\": \"RH\"}\n      ]\n    }\n  ],\n  \"current_model\": \"Toxoplasma gondii Rab11A is a small GTPase that controls vesicular trafficking at multiple stages of the parasite life cycle: it mediates delivery of new plasma membrane and IMC components to daughter cells during cytokinesis (interacting with the MTIP/MLC1–MyosinA motor complex), drives cytoskeleton-dependent constitutive transport and apically polarized secretion of dense granules (regulating MIC2 release and host cell invasion), and is phosphorylated by TgCDPK7 to regulate intracellular development and protein trafficking; on the host side, Toxoplasma selectively sequesters host Rab11A-positive vesicles into the parasitophorous vacuole intravacuolar network in a TgGRA2/GRA6-dependent manner, where TgLCAT mediates their lipolytic processing for nutrient acquisition.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"Toxoplasma gondii Rab11A is a small GTPase that governs vesicular trafficking at multiple stages of the parasite life cycle by directing the delivery of membrane and secretory cargo to specific cellular destinations [#0, #4]. During cytokinesis it supplies new plasma membrane to nascent daughter cells, and its loss produces daughters with incompletely formed Inner Membrane Complex and a block at a late stage of division; this membrane traffic is coupled to the glideosome motor through physical association with MTIP/MLC1, implicating an MTIP-Myosin motor in Rab11A-driven IMC maturation [#0, #1]. Rab11A additionally drives cytoskeleton-dependent transport and constitutive secretion of dense granules, regulates transmembrane protein delivery to the plasma membrane during replication, and mediates apically polarized delivery that controls MIC2 secretion and early secretory events during host cell invasion [#4, #5, #6]. Its activity is regulated by phosphorylation by TgCDPK7, which is critical for intracellular development and protein trafficking, and by the ER-localized TBC-domain GAP TBC9, which physically interacts with Rab11A and functions in early vesicular transport and daughter IMC formation [#7, #9]. On the host side, Toxoplasma sequesters host Rab11A-positive class I FIP vesicles into the parasitophorous vacuole intravacuolar network in a TgGRA2/GRA6-dependent manner, where they are lipolytically processed by the phospholipase TgLCAT, and host autophagy limits parasite access to these vesicles to restrict replication [#2, #3, #8, #10].\",\n  \"teleology\": [\n    {\n      \"year\": 2009,\n      \"claim\": \"Established the first cellular role for apicomplexan Rab11A by asking what drives membrane delivery during parasite cell division, showing it is required to build daughter cell surfaces.\",\n      \"evidence\": \"Dominant-negative/ablation of Rab11A with live imaging and phenotypic readout of IMC formation and cytokinesis, plus association with the MTIP/MLC1 glideosome component\",\n      \"pmids\": [\"19165333\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct GTPase cycle/effector biochemistry not resolved\", \"MTIP association shown by a single co-IP without reciprocal validation\", \"Vesicle cargo composition not defined\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}, {\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Extended Rab11A biology to the host-parasite interface by asking whether Toxoplasma exploits host Rab11A vesicles, demonstrating selective sequestration into the PV and their lipolytic processing.\",\n      \"evidence\": \"Correlative light/EM plus genetic deletion of gra2/gra6 and TgLCAT overexpression with quantitative microscopy of intra-PV GFP-Rab11A vesicles\",\n      \"pmids\": [\"29070609\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which GRA2/GRA6 stabilize tubules for capture not resolved\", \"TgLCAT-Rab11A degradation link rests on single overexpression experiment\", \"Nutrient/cargo actually acquired not directly identified\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Defined the secretory scope of parasite Rab11A by asking which trafficking events it controls, revealing roles in dense granule transport/secretion, transmembrane protein delivery, and apically polarized MIC2 secretion during invasion.\",\n      \"evidence\": \"Loss-of-function of Rab11A with live imaging of dense granule and Rab11A-GFP dynamics plus secretion assays including MIC2\",\n      \"pmids\": [\"32463830\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Effectors linking Rab11A to the cytoskeleton not identified\", \"How apical polarization of Rab11A vesicles is established is unknown\", \"Direct versus indirect effect on MIC2 secretion not dissected\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Addressed how parasite Rab11A activity is regulated by identifying it as a substrate of TgCDPK7, linking a kinase signal to trafficking and intracellular development.\",\n      \"evidence\": \"Quantitative phosphoproteomics of TgCDPK7 knockout with functional validation of phosphosite importance\",\n      \"pmids\": [\"33635921\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of the specific phosphorylation on Rab11A GTPase activity not defined\", \"Whether phosphorylation alters effector binding unknown\", \"Single-study phosphoproteomic identification\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Refined the host vesicle recruitment mechanism by asking which host Rab11A vesicle subclass is captured, showing selectivity for class I FIP (FIP5) but not class II FIP (FIP3) vesicles, and demonstrating host autophagy restricts parasite access.\",\n      \"evidence\": \"Host Rab11A depletion with quantitative microscopy of intra-PV FIP vesicles and dominant-negative FIP3 mutants (MBoC); CRISPR knockout of host atg5/rab11a with replication assays and qRT-PCR (Front Microbiol)\",\n      \"pmids\": [\"35274991\", \"36532461\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Parasite receptor/factor that discriminates FIP subclasses not identified\", \"Mechanistic link between autophagy and Rab11A vesicle availability unresolved\", \"Studies focus on host Rab11A, not parasite Rab11A\"],\n      \"attribution\": [{\"species\": \"Homo sapiens\", \"strain\": \"\"}, {\"species\": \"Homo sapiens\", \"strain\": \"HeLa\"}, {\"species\": \"Toxoplasma gondii\", \"strain\": \"RH\"}]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identified a candidate regulatory GAP by asking what controls the parasite Rab11A GTPase cycle, showing the ER-localized TBC9 interacts with Rab11A and functions in early transport and daughter IMC formation.\",\n      \"evidence\": \"Pull-down assay of TBC9 with Rab11A/Rab11B/Rab2, fluorescence localization, and TBC9 loss-of-function phenotyping\",\n      \"pmids\": [\"38762629\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"GAP activity on Rab11A not biochemically demonstrated\", \"Interaction shown by a single pull-down without reciprocal validation\", \"TBC9 binds multiple Rabs, so Rab11A specificity of its function is unclear\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How Rab11A GTPase cycling, phosphorylation, and GAP regulation are integrated with its distinct effector complexes at each trafficking step remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural or biochemical map of Rab11A GEF/GAP/effector cycle in the parasite\", \"Direct effectors linking Rab11A to cytoskeletal transport not identified\", \"Consequences of TgCDPK7 phosphorylation on nucleotide state and effector engagement unknown\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GTPase activity\", \"supporting_discovery_ids\": [0, 4, 9], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]},\n      {\"term_id\": \"small GTPase-mediated vesicular transport\", \"supporting_discovery_ids\": [0, 4, 5, 6], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]},\n      {\"term_id\": \"protein kinase substrate (phosphorylation by TgCDPK7)\", \"supporting_discovery_ids\": [7], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"cytoplasmic vesicle / secretory vesicle\", \"supporting_discovery_ids\": [4, 5, 6], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]},\n      {\"term_id\": \"apical pole of parasite\", \"supporting_discovery_ids\": [6], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]},\n      {\"term_id\": \"host parasitophorous vacuole intravacuolar network\", \"supporting_discovery_ids\": [2, 3, 8], \"attribution\": [{\"species\": \"Homo sapiens\", \"strain\": \"\"}]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"cytokinesis / daughter cell IMC biogenesis\", \"supporting_discovery_ids\": [0, 1, 9], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]},\n      {\"term_id\": \"dense granule constitutive secretion\", \"supporting_discovery_ids\": [4], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]},\n      {\"term_id\": \"regulated microneme (MIC2) secretion / host cell invasion\", \"supporting_discovery_ids\": [6], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]},\n      {\"term_id\": \"host vesicle sequestration and nutrient acquisition\", \"supporting_discovery_ids\": [2, 3, 8, 10], \"attribution\": [{\"species\": \"Homo sapiens\", \"strain\": \"\"}, {\"species\": \"Toxoplasma gondii\", \"strain\": \"RH\"}]}\n    ],\n    \"complexes\": [\n      \"MTIP/MLC1-MyosinA glideosome motor association\"\n    ],\n    \"partners\": [\n      \"MTIP/MLC1 (glideosome, T. gondii)\",\n      \"TgCDPK7 (kinase, T. gondii)\",\n      \"TBC9 (ER-localized TBC-domain Rab GAP, T. gondii)\",\n      \"TgGRA2 / TgGRA6 (host vesicle sequestration, T. gondii)\",\n      \"TgLCAT (phospholipase, T. gondii)\",\n      \"FIP5 / class I FIP vesicles (host)\"\n    ],\n    \"other_free_text\": [\n      \"Host Rab11A-positive class I FIP (FIP5) vesicles are selectively recruited to the PV; class II FIP (FIP3) vesicles are not.\",\n      \"Host autophagy (ATG5-dependent) limits parasite access to Rab11A vesicles and restricts replication; rab11a disruption increases parasite gra2 transcription.\",\n      \"TBC9 interacts with Rab11A, Rab11B, and Rab2, so its regulatory function is not exclusive to Rab11A.\"\n    ]\n  },\n  \"strain_conflicts\": []\n}","audit_flag":null,"strains":[{"strain":"ME49","gene_id":"TGME49_289680","name":"RAB11A","product":"ras-related protein RAB11A","uniprot_acc":"A0A125YY17","chromosome":"TGME49_chrIX","start_pos":3372902},{"strain":"GT1","gene_id":"TGGT1_289680","name":null,"product":"Ras-related protein Rab11","uniprot_acc":null,"chromosome":"TGGT1_chrIX","start_pos":3181746},{"strain":"RH88","gene_id":"TGRH88_014430","name":null,"product":"Ras-related protein Rab11","uniprot_acc":null,"chromosome":"CM023090","start_pos":3437879},{"strain":"VEG","gene_id":"TGVEG_289680","name":null,"product":"Ras-related protein Rab11","uniprot_acc":"A0A0F7V7A6","chromosome":"TGVEG_chrIX","start_pos":3407739}]}