{"gene":"TG_311470","gene_symbol":"RON5","run_date":"2026-09-08T15:15:13","timeline":{"discoveries":[{"year":2009,"finding":"RON5 is a component of the moving junction (MJ) complex, localizing to the cytoplasmic face of the host plasma membrane during invasion, as demonstrated by differential permeabilization and loading of RON-specific antibodies into the host cell.","method":"Differential permeabilization, immunoprecipitation, metabolic labeling","journal":"PLoS pathogens","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal co-IP and differential permeabilization, independently replicated across multiple labs","pmids":["19247437"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2008,"finding":"RON5 is conserved in Plasmodium and co-immunoprecipitates with RON8 and other known MJ proteins (RON2, RON4) from extracellular parasites, indicating RON5 is part of a preformed complex within the parasite prior to secretion.","method":"Co-immunoprecipitation","journal":"Cellular microbiology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — single Co-IP but corroborated by multiple subsequent studies","pmids":["19134112"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2014,"finding":"Conditional knockdown of RON5 results in complete degradation of RON2 and mistargeting of RON4 within the parasite secretory pathway, demonstrating that RON5 is essential for organization and stability of the MJ RON complex.","method":"Conditional knockdown (tetracycline-regulatable), Western blot, immunofluorescence","journal":"PLoS pathogens","confidence":"High","confidence_rationale":"Tier 2 / Strong — conditional knockdown with multiple orthogonal readouts (protein stability, localization, functional invasion assays) in a single rigorous study","pmids":["24651769"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2014,"finding":"Loss of RON5 by conditional knockdown renders parasites unable to invade host cells, providing the first genetic demonstration that RON5 is required for host cell penetration.","method":"Conditional knockdown, invasion assay","journal":"PLoS pathogens","confidence":"High","confidence_rationale":"Tier 2 / Strong — conditional genetic knockdown with defined invasion phenotype, multiple orthogonal methods in single study","pmids":["24651769"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2014,"finding":"Parasites lacking RON5 fail to form evacuoles, demonstrating that an intact MJ complex (dependent on RON5) is a prerequisite for secretion of rhoptry bulb contents.","method":"Conditional knockdown, evacuole formation assay","journal":"PLoS pathogens","confidence":"High","confidence_rationale":"Tier 2 / Strong — conditional knockdown with specific functional readout (evacuole formation) alongside invasion assays","pmids":["24651769"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2014,"finding":"Disruption of the MJ complex by RON5 depletion does not impact parasite egress, indicating RON5 and the MJ complex are not required for egress.","method":"Conditional knockdown, egress assay","journal":"PLoS pathogens","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — single lab, single method for this specific negative finding, though rigorously performed","pmids":["24651769"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2014,"finding":"Functional complementation of the RON5 conditional mutant revealed that proteolytic separation of RON5 N- and C-terminal fragments is dispensable, but a portion of the C-terminal domain is critical for RON2 stability and invasion function.","method":"Functional complementation, mutagenesis, invasion assay, Western blot","journal":"PLoS pathogens","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — complementation with domain deletion mutagenesis and defined functional readout in single rigorous study","pmids":["24651769"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2017,"finding":"RON5 cooperates with RON2 and RON4 to actively recruit host proteins CIN85, CD2AP, and ESCRT-I components ALIX and TSG101 to the moving junction during invasion, with the parasite mimicking conserved host binding interfaces.","method":"Co-immunoprecipitation, interaction mapping, parasite mutant analysis (invasion and virulence assays)","journal":"Nature microbiology","confidence":"High","confidence_rationale":"Tier 2 / Strong — interaction mapping with multiple orthogonal methods, mutant parasites with defined phenotypes in culture and in vivo","pmids":["28848228"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2017,"finding":"RON4 knockout decreases the expression of the endogenous N-terminal fragment of RON5, indicating that RON4 contributes to the stability of RON5 within the MJ complex.","method":"CRISPR-Cas9 knockout, Western blot","journal":"Parasitology","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — CRISPR knockout with defined protein-level readout, single lab, single method","pmids":["28662729"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":"RH"}]},{"year":2017,"finding":"RON4L1 traffic to the MJ is not affected in the absence of RON2, RON4, or RON5, suggesting that RON4L1 is part of an independent MJ sub-complex rather than depending on these RON proteins for its targeting.","method":"Knockout cell line generation, immunofluorescence localization during invasion","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — knockout lines with localization assays; implicates RON5 negatively in RON4L1 trafficking, single lab","pmids":["29263399"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]}],"current_model":"RON5 is a rhoptry neck protein that forms a preformed complex with RON2, RON4, and RON8 inside the parasite, is secreted and targeted to the cytoplasmic face of the host plasma membrane during invasion to constitute the moving junction (MJ); RON5 is essential for stabilizing RON2 (and RON4) within the secretory pathway, for host cell penetration, and for rhoptry bulb secretion (evacuole formation), while its C-terminal domain is specifically required for RON2 stability and invasion function, and together with RON2 and RON4 it subverts host proteins (CIN85, CD2AP, ALIX, TSG101) to enable efficient invasion."},"narrative":{"mechanistic_narrative":"RON5 is a rhoptry neck protein of Toxoplasma gondii that functions as a core structural component of the moving junction (MJ), the ring-shaped interface through which the parasite penetrates host cells [PMID:19247437]. Within the parasite it assembles into a preformed complex with the other RON proteins (RON2, RON4, RON8) prior to secretion, and this complex is discharged and targeted to the cytoplasmic face of the host plasma membrane during invasion [PMID:19247437, PMID:19134112]. RON5 acts as an organizational hub for this complex: its loss triggers complete degradation of RON2 and mistargeting of RON4 within the secretory pathway, with a portion of its C-terminal domain being specifically required for RON2 stability and invasion function [PMID:24651769]. Functionally, RON5 is essential for host cell penetration and for secretion of rhoptry bulb contents (evacuole formation), but is dispensable for parasite egress [PMID:24651769]. Together with RON2 and RON4, RON5 actively recruits host proteins CIN85, CD2AP, and the ESCRT-I components ALIX and TSG101 to the MJ by mimicking conserved host binding interfaces, coupling junction assembly to host machinery during invasion [PMID:28848228].","teleology":[{"year":2008,"claim":"Establishing that RON5 physically belongs to the MJ RON machinery was the first step in defining its role, showing it is part of a preformed intracellular complex rather than an isolated factor.","evidence":"Co-immunoprecipitation of RON5 with RON8, RON2, and RON4 from extracellular parasites","pmids":["19134112"],"confidence":"Medium","gaps":["Does not establish stoichiometry or direct binding partners within the complex","Does not address function during invasion"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2009,"claim":"Localizing RON5 to the cytoplasmic face of the host plasma membrane during invasion demonstrated it is a bona fide MJ component secreted into the host interface, not merely a parasite-resident protein.","evidence":"Differential permeabilization with RON-specific antibodies, immunoprecipitation, and metabolic labeling","pmids":["19247437"],"confidence":"High","gaps":["Does not resolve how RON5 is anchored at the host membrane","Functional requirement not yet tested genetically"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2014,"claim":"Conditional knockdown established RON5 as an organizational keystone of the MJ complex and defined its functional requirements, resolving whether it is structural or merely associated.","evidence":"Tetracycline-regulatable knockdown with Western blot, immunofluorescence, invasion, evacuole, and egress assays; complementation with domain mutagenesis","pmids":["24651769"],"confidence":"High","gaps":["Mechanism by which RON5 stabilizes RON2 within the secretory pathway is not defined","Molecular function of the critical C-terminal region is unresolved","Reason egress is spared while invasion fails is not mechanistically explained"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2017,"claim":"Mapping host-protein recruitment showed the RON5/RON2/RON4 complex subverts host CIN85, CD2AP, and ESCRT-I components by mimicking conserved binding interfaces, connecting MJ assembly to host cellular machinery.","evidence":"Co-immunoprecipitation, interaction mapping, and parasite mutant analysis in culture and in vivo","pmids":["28848228"],"confidence":"High","gaps":["The specific contribution of RON5 versus RON2/RON4 to each host interaction is not isolated","Downstream consequences of host protein recruitment for invasion are not fully defined"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2017,"claim":"Reciprocal dependency and complex-boundary experiments refined the interdependence of RON components, showing RON4 supports RON5 N-terminal stability while RON4L1 traffics independently of RON5.","evidence":"CRISPR-Cas9 knockout and Western blot for RON4 dependency; knockout lines with invasion-stage immunofluorescence for RON4L1 trafficking","pmids":["28662729","29263399"],"confidence":"Medium","gaps":["RON4-mediated stabilization of RON5 shown only at protein level in RH strain","Whether RON4L1 forms a distinct MJ sub-complex is not directly demonstrated"],"attribution":[{"species":"Toxoplasma gondii","strain":"RH"},{"species":"Toxoplasma gondii","strain":""}]},{"year":null,"claim":"The structural basis of RON5-mediated RON2 stabilization and the molecular identity of its critical C-terminal function within the secretory pathway remain unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model of the RON5-RON2 interaction","Mechanism of C-terminal domain action on RON2 stability undefined"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]}],"mechanism_profile":{"molecular_activity":[],"localization":[],"pathway":[],"complexes":["Moving junction RON complex (RON2, RON4, RON5, RON8)"],"partners":["RON2","RON4","RON8","HOST CIN85","HOST CD2AP","HOST ALIX","HOST TSG101"],"other_free_text":["RON5 loss causes complete degradation of RON2 and mistargeting of RON4 in the secretory pathway [#2].","A portion of the RON5 C-terminal domain is specifically required for RON2 stability and invasion; proteolytic separation of N- and C-terminal fragments is dispensable [#6].","RON5 is required for host cell penetration and for rhoptry bulb secretion (evacuole formation) but not for egress [#3, #4, #5].","RON4 contributes to stability of the endogenous RON5 N-terminal fragment (shown in RH strain) [#8].","RON4L1 traffics to the MJ independently of RON2, RON4, or RON5 [#9]."]},"strain_conflicts":[]},"prefetch_data":{},"corpus_meta":[{"pmid":"19247437","id":"PMC_19247437","title":"Export of a Toxoplasma gondii rhoptry neck protein complex at the host cell membrane to form the moving junction during invasion.","date":"2009","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/19247437","citation_count":238,"is_preprint":false},{"pmid":"19134112","id":"PMC_19134112","title":"Novel components of the Apicomplexan moving junction reveal conserved and coccidia-restricted elements.","date":"2008","source":"Cellular microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/19134112","citation_count":92,"is_preprint":false},{"pmid":"28848228","id":"PMC_28848228","title":"Efficient invasion by Toxoplasma depends on the subversion of host protein networks.","date":"2017","source":"Nature microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/28848228","citation_count":50,"is_preprint":false},{"pmid":"24651769","id":"PMC_24651769","title":"RON5 is critical for organization and function of the Toxoplasma moving junction complex.","date":"2014","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/24651769","citation_count":47,"is_preprint":false},{"pmid":"29263399","id":"PMC_29263399","title":"RON4L1 is a new member of the moving junction complex in Toxoplasma gondii.","date":"2017","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/29263399","citation_count":16,"is_preprint":false},{"pmid":"26821295","id":"PMC_26821295","title":"Protective efficacy of pVAX-RON5p against acute and chronic infections of Toxoplasma gondii in BALB/c mice.","date":"2016","source":"Experimental parasitology","url":"https://pubmed.ncbi.nlm.nih.gov/26821295","citation_count":10,"is_preprint":false},{"pmid":"28662729","id":"PMC_28662729","title":"The moving junction protein RON4, although not critical, facilitates host cell invasion and stabilizes MJ members.","date":"2017","source":"Parasitology","url":"https://pubmed.ncbi.nlm.nih.gov/28662729","citation_count":6,"is_preprint":false},{"pmid":"30326220","id":"PMC_30326220","title":"Deciphering ophthalmic adaptive inhibitors targeting RON4 of Toxoplasma gondii: An integrative in silico approach.","date":"2018","source":"Life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/30326220","citation_count":5,"is_preprint":false},{"pmid":"40881926","id":"PMC_40881926","title":"In silico evaluation of Toxoplasma gondii rhoptry neck proteins (TgRONs) for potential immunogenic epitopes.","date":"2025","source":"EXCLI journal","url":"https://pubmed.ncbi.nlm.nih.gov/40881926","citation_count":1,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":6563,"output_tokens":2189,"usd":0.026262,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":7626,"output_tokens":2701,"usd":0.052827,"stage2_stop_reason":"end_turn"},"total_usd":0.079089,"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\": 2009,\n      \"finding\": \"RON5 is a component of the moving junction (MJ) complex, localizing to the cytoplasmic face of the host plasma membrane during invasion, as demonstrated by differential permeabilization and loading of RON-specific antibodies into the host cell.\",\n      \"method\": \"Differential permeabilization, immunoprecipitation, metabolic labeling\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal co-IP and differential permeabilization, independently replicated across multiple labs\",\n      \"pmids\": [\"19247437\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"RON5 is conserved in Plasmodium and co-immunoprecipitates with RON8 and other known MJ proteins (RON2, RON4) from extracellular parasites, indicating RON5 is part of a preformed complex within the parasite prior to secretion.\",\n      \"method\": \"Co-immunoprecipitation\",\n      \"journal\": \"Cellular microbiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — single Co-IP but corroborated by multiple subsequent studies\",\n      \"pmids\": [\"19134112\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Conditional knockdown of RON5 results in complete degradation of RON2 and mistargeting of RON4 within the parasite secretory pathway, demonstrating that RON5 is essential for organization and stability of the MJ RON complex.\",\n      \"method\": \"Conditional knockdown (tetracycline-regulatable), Western blot, immunofluorescence\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conditional knockdown with multiple orthogonal readouts (protein stability, localization, functional invasion assays) in a single rigorous study\",\n      \"pmids\": [\"24651769\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Loss of RON5 by conditional knockdown renders parasites unable to invade host cells, providing the first genetic demonstration that RON5 is required for host cell penetration.\",\n      \"method\": \"Conditional knockdown, invasion assay\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conditional genetic knockdown with defined invasion phenotype, multiple orthogonal methods in single study\",\n      \"pmids\": [\"24651769\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Parasites lacking RON5 fail to form evacuoles, demonstrating that an intact MJ complex (dependent on RON5) is a prerequisite for secretion of rhoptry bulb contents.\",\n      \"method\": \"Conditional knockdown, evacuole formation assay\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conditional knockdown with specific functional readout (evacuole formation) alongside invasion assays\",\n      \"pmids\": [\"24651769\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Disruption of the MJ complex by RON5 depletion does not impact parasite egress, indicating RON5 and the MJ complex are not required for egress.\",\n      \"method\": \"Conditional knockdown, egress assay\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — single lab, single method for this specific negative finding, though rigorously performed\",\n      \"pmids\": [\"24651769\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Functional complementation of the RON5 conditional mutant revealed that proteolytic separation of RON5 N- and C-terminal fragments is dispensable, but a portion of the C-terminal domain is critical for RON2 stability and invasion function.\",\n      \"method\": \"Functional complementation, mutagenesis, invasion assay, Western blot\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — complementation with domain deletion mutagenesis and defined functional readout in single rigorous study\",\n      \"pmids\": [\"24651769\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"RON5 cooperates with RON2 and RON4 to actively recruit host proteins CIN85, CD2AP, and ESCRT-I components ALIX and TSG101 to the moving junction during invasion, with the parasite mimicking conserved host binding interfaces.\",\n      \"method\": \"Co-immunoprecipitation, interaction mapping, parasite mutant analysis (invasion and virulence assays)\",\n      \"journal\": \"Nature microbiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — interaction mapping with multiple orthogonal methods, mutant parasites with defined phenotypes in culture and in vivo\",\n      \"pmids\": [\"28848228\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"RON4 knockout decreases the expression of the endogenous N-terminal fragment of RON5, indicating that RON4 contributes to the stability of RON5 within the MJ complex.\",\n      \"method\": \"CRISPR-Cas9 knockout, Western blot\",\n      \"journal\": \"Parasitology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — CRISPR knockout with defined protein-level readout, single lab, single method\",\n      \"pmids\": [\"28662729\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"RH\"}]\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"RON4L1 traffic to the MJ is not affected in the absence of RON2, RON4, or RON5, suggesting that RON4L1 is part of an independent MJ sub-complex rather than depending on these RON proteins for its targeting.\",\n      \"method\": \"Knockout cell line generation, immunofluorescence localization during invasion\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — knockout lines with localization assays; implicates RON5 negatively in RON4L1 trafficking, single lab\",\n      \"pmids\": [\"29263399\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    }\n  ],\n  \"current_model\": \"RON5 is a rhoptry neck protein that forms a preformed complex with RON2, RON4, and RON8 inside the parasite, is secreted and targeted to the cytoplasmic face of the host plasma membrane during invasion to constitute the moving junction (MJ); RON5 is essential for stabilizing RON2 (and RON4) within the secretory pathway, for host cell penetration, and for rhoptry bulb secretion (evacuole formation), while its C-terminal domain is specifically required for RON2 stability and invasion function, and together with RON2 and RON4 it subverts host proteins (CIN85, CD2AP, ALIX, TSG101) to enable efficient invasion.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"RON5 is a rhoptry neck protein of Toxoplasma gondii that functions as a core structural component of the moving junction (MJ), the ring-shaped interface through which the parasite penetrates host cells [#0]. Within the parasite it assembles into a preformed complex with the other RON proteins (RON2, RON4, RON8) prior to secretion, and this complex is discharged and targeted to the cytoplasmic face of the host plasma membrane during invasion [#0, #1]. RON5 acts as an organizational hub for this complex: its loss triggers complete degradation of RON2 and mistargeting of RON4 within the secretory pathway, with a portion of its C-terminal domain being specifically required for RON2 stability and invasion function [#2, #6]. Functionally, RON5 is essential for host cell penetration and for secretion of rhoptry bulb contents (evacuole formation), but is dispensable for parasite egress [#3, #4, #5]. Together with RON2 and RON4, RON5 actively recruits host proteins CIN85, CD2AP, and the ESCRT-I components ALIX and TSG101 to the MJ by mimicking conserved host binding interfaces, coupling junction assembly to host machinery during invasion [#7].\",\n  \"teleology\": [\n    {\n      \"year\": 2008,\n      \"claim\": \"Establishing that RON5 physically belongs to the MJ RON machinery was the first step in defining its role, showing it is part of a preformed intracellular complex rather than an isolated factor.\",\n      \"evidence\": \"Co-immunoprecipitation of RON5 with RON8, RON2, and RON4 from extracellular parasites\",\n      \"pmids\": [\"19134112\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Does not establish stoichiometry or direct binding partners within the complex\", \"Does not address function during invasion\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Localizing RON5 to the cytoplasmic face of the host plasma membrane during invasion demonstrated it is a bona fide MJ component secreted into the host interface, not merely a parasite-resident protein.\",\n      \"evidence\": \"Differential permeabilization with RON-specific antibodies, immunoprecipitation, and metabolic labeling\",\n      \"pmids\": [\"19247437\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Does not resolve how RON5 is anchored at the host membrane\", \"Functional requirement not yet tested genetically\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Conditional knockdown established RON5 as an organizational keystone of the MJ complex and defined its functional requirements, resolving whether it is structural or merely associated.\",\n      \"evidence\": \"Tetracycline-regulatable knockdown with Western blot, immunofluorescence, invasion, evacuole, and egress assays; complementation with domain mutagenesis\",\n      \"pmids\": [\"24651769\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which RON5 stabilizes RON2 within the secretory pathway is not defined\", \"Molecular function of the critical C-terminal region is unresolved\", \"Reason egress is spared while invasion fails is not mechanistically explained\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Mapping host-protein recruitment showed the RON5/RON2/RON4 complex subverts host CIN85, CD2AP, and ESCRT-I components by mimicking conserved binding interfaces, connecting MJ assembly to host cellular machinery.\",\n      \"evidence\": \"Co-immunoprecipitation, interaction mapping, and parasite mutant analysis in culture and in vivo\",\n      \"pmids\": [\"28848228\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"The specific contribution of RON5 versus RON2/RON4 to each host interaction is not isolated\", \"Downstream consequences of host protein recruitment for invasion are not fully defined\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Reciprocal dependency and complex-boundary experiments refined the interdependence of RON components, showing RON4 supports RON5 N-terminal stability while RON4L1 traffics independently of RON5.\",\n      \"evidence\": \"CRISPR-Cas9 knockout and Western blot for RON4 dependency; knockout lines with invasion-stage immunofluorescence for RON4L1 trafficking\",\n      \"pmids\": [\"28662729\", \"29263399\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"RON4-mediated stabilization of RON5 shown only at protein level in RH strain\", \"Whether RON4L1 forms a distinct MJ sub-complex is not directly demonstrated\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"RH\"}, {\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The structural basis of RON5-mediated RON2 stabilization and the molecular identity of its critical C-terminal function within the secretory pathway remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model of the RON5-RON2 interaction\", \"Mechanism of C-terminal domain action on RON2 stability undefined\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0005515\", \"supporting_discovery_ids\": [1, 2, 7], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0020003\", \"supporting_discovery_ids\": [0], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]},\n      {\"term_id\": \"GO:0009986\", \"supporting_discovery_ids\": [0], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0044409\", \"supporting_discovery_ids\": [3, 7], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]}\n    ],\n    \"complexes\": [\n      \"Moving junction RON complex (RON2, RON4, RON5, RON8)\"\n    ],\n    \"partners\": [\n      \"RON2\",\n      \"RON4\",\n      \"RON8\",\n      \"Host CIN85\",\n      \"Host CD2AP\",\n      \"Host ALIX\",\n      \"Host TSG101\"\n    ],\n    \"other_free_text\": [\n      \"RON5 loss causes complete degradation of RON2 and mistargeting of RON4 in the secretory pathway [#2].\",\n      \"A portion of the RON5 C-terminal domain is specifically required for RON2 stability and invasion; proteolytic separation of N- and C-terminal fragments is dispensable [#6].\",\n      \"RON5 is required for host cell penetration and for rhoptry bulb secretion (evacuole formation) but not for egress [#3, #4, #5].\",\n      \"RON4 contributes to stability of the endogenous RON5 N-terminal fragment (shown in RH strain) [#8].\",\n      \"RON4L1 traffics to the MJ independently of RON2, RON4, or RON5 [#9].\"\n    ]\n  },\n  \"strain_conflicts\": []\n}\n```","audit_flag":null,"strains":[{"strain":"ME49","gene_id":"TGME49_311470","name":"RON5","product":"rhoptry neck protein RON5","uniprot_acc":"S8ERS3","chromosome":"TGME49_chrXI","start_pos":2083415},{"strain":"GT1","gene_id":"TGGT1_311470","name":null,"product":"rhoptry neck protein RON5","uniprot_acc":null,"chromosome":"TGGT1_chrXI","start_pos":2047692},{"strain":"RH88","gene_id":"TGRH88_050950","name":null,"product":"rhoptry neck protein RON5","uniprot_acc":null,"chromosome":"CM023092","start_pos":2132984},{"strain":"VEG","gene_id":"TGVEG_311470","name":null,"product":"rhoptry neck protein RON5","uniprot_acc":"A0A125YSX9","chromosome":"TGVEG_chrXI","start_pos":2095996}]}