{"gene":"PF3D7_1441800","run_date":"2026-09-08T14:19:07","timeline":{"discoveries":[{"year":2021,"finding":"PF3D7_1441800 (VPS60/PfVps60) is a component of the P. falciparum ESCRT-III machinery and participates in extracellular vesicle production. Disruption of the PfVps60 gene reduced the number of EVs produced and affected the distribution of other ESCRT-III components. Purified recombinant PfVps60 protein, together with PfVps32 and PfBro1, reconstituted multivesicular body formation and membrane shedding in giant unilamellar vesicles. PfVps60 was confirmed present in EVs purified from parasitized RBC cultures by super-resolution microscopy and dot blot assays.","method":"Genetic knockout, in vitro reconstitution with purified recombinant proteins in giant unilamellar vesicles, super-resolution microscopy, dot blot","journal":"PLoS pathogens","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — knockout phenotype combined with in vitro reconstitution using purified recombinant proteins, plus direct detection by super-resolution microscopy; multiple orthogonal methods in a single study","pmids":["33798247"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2021,"finding":"The P. falciparum ESCRT-III sub-complex is recruited via an alternative pathway involving PfBro1 acting upstream of PfVps32 and PfVps60, differing from canonical eukaryotic ESCRT recruitment.","method":"Functional reconstitution with purified recombinant proteins, genetic knockout analysis","journal":"PLoS pathogens","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional reconstitution supports pathway order but mechanistic detail of recruitment order inferred from reconstitution data in a single study","pmids":["33798247"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2026,"finding":"In P. falciparum 3D7, VPS60 (PF3D7_1441800) knockout impaired EV-mediated export of chaperones PfHsp70-x and PfHsp110 under heat shock conditions, altered protein aggregate dynamics (inducing redistribution of aggregates near the parasitophorous vacuole), reduced induction of cytosolic chaperone PfHsp70-1, and resulted in early loss of parasite viability following heat shock. Supplementation of PfVps60KO parasites with EVs from heat-stressed wild-type parasites partially rescued heat shock tolerance in a dose-dependent manner, linking ESCRT-dependent vesicular trafficking to proteostasis.","method":"Genetic knockout (PfVps60KO line), proteomic profiling, fluorescence microscopy (aggregate distribution), viability assays, EV supplementation rescue experiment","journal":"Emerging microbes & infections","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean knockout with multiple orthogonal readouts (proteomics, microscopy, viability, rescue) in a single lab study","pmids":["42253101"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":"3D7"}]}],"current_model":"PF3D7_1441800 (VPS60) is a P. falciparum ESCRT-III subunit recruited via PfBro1 that drives extracellular vesicle biogenesis (multivesicular body formation and membrane shedding), is itself packaged into EVs, and mediates EV-dependent export of chaperones (PfHsp70-x, PfHsp110) required for proteostasis and parasite survival under febrile heat-shock conditions."},"narrative":{"mechanistic_narrative":"PF3D7_1441800 (VPS60) is a component of the Plasmodium falciparum ESCRT-III machinery that drives extracellular vesicle (EV) biogenesis and links vesicular trafficking to parasite proteostasis [PMID:33798247, PMID:42253101]. Genetic disruption of VPS60 reduces EV production and perturbs the distribution of other ESCRT-III components, and purified recombinant PfVps60 acts together with PfVps32 and PfBro1 to reconstitute multivesicular body formation and membrane shedding in giant unilamellar vesicles; PfVps60 is itself detected in EVs purified from parasitized red blood cell cultures [PMID:33798247]. Recruitment of this ESCRT-III sub-complex proceeds through a non-canonical route in which PfBro1 acts upstream of PfVps32 and PfVps60 [PMID:33798247]. Functionally, VPS60 is required for EV-mediated export of the chaperones PfHsp70-x and PfHsp110 under heat shock, and its loss alters protein aggregate dynamics, dampens induction of cytosolic PfHsp70-1, and causes early loss of parasite viability after heat shock, a defect partially rescued by supplying EVs from heat-stressed wild-type parasites [PMID:42253101].","teleology":[{"year":2021,"claim":"Whether PF3D7_1441800 functions in the parasite's ESCRT machinery and contributes to vesicle formation was unknown; establishing this defined its role in EV biogenesis.","evidence":"Genetic knockout reducing EV number, in vitro reconstitution of MVB formation and membrane shedding in giant unilamellar vesicles with recombinant PfVps60/PfVps32/PfBro1, super-resolution microscopy and dot blot detection in EVs","pmids":["33798247"],"confidence":"High","gaps":["Structural basis of PfVps60 membrane remodeling not resolved","Stoichiometry and assembly geometry of the reconstituted complex not defined","Cargo selectivity of VPS60-dependent EVs not addressed in this study"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2021,"claim":"How the ESCRT-III sub-complex including VPS60 is recruited was unclear; ordering PfBro1 upstream of PfVps32 and PfVps60 revealed a non-canonical recruitment pathway.","evidence":"Functional reconstitution with purified recombinant proteins combined with genetic knockout analysis","pmids":["33798247"],"confidence":"Medium","gaps":["Recruitment order inferred from reconstitution rather than direct in-cell temporal measurement","Molecular interface between PfBro1 and PfVps60 not mapped","Whether additional adaptors participate in recruitment is unknown"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2026,"claim":"Whether VPS60-dependent EVs serve a physiological function was open; linking VPS60 loss to failed chaperone export and heat-shock survival established a role in EV-dependent proteostasis.","evidence":"PfVps60KO line in P. falciparum 3D7 with proteomic profiling, aggregate-distribution microscopy, viability assays, and dose-dependent EV supplementation rescue","pmids":["42253101"],"confidence":"Medium","gaps":["Molecular mechanism by which chaperones are loaded into VPS60-dependent EVs is unknown","How exported EV chaperones act on recipient cells or the parasite surface is undefined","Only partial rescue achieved, leaving other VPS60-dependent functions unaccounted for"],"attribution":[{"species":"Plasmodium falciparum","strain":"3D7"}]},{"year":null,"claim":"The structural mechanism of PfVps60-driven membrane remodeling and the basis of chaperone cargo selection into EVs remain unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model of PfVps60 assembly on membranes","Cargo-sorting mechanism directing PfHsp70-x/PfHsp110 into EVs unknown","Regulation of VPS60 activity under febrile stress not defined"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]}],"mechanism_profile":{"molecular_activity":[],"localization":[],"pathway":[],"complexes":["ESCRT-III (P. falciparum, with PfVps32 and PfBro1)"],"partners":["PFVPS32","PFBRO1","PFHSP70-X (EV CARGO)","PFHSP110 (EV CARGO)"],"other_free_text":["VPS60 loss redistributes protein aggregates near the parasitophorous vacuole and dampens induction of cytosolic PfHsp70-1 [#2]","PfVps60 is itself packaged into extracellular vesicles released from parasitized red blood cells [#0]","EVs from heat-stressed wild-type parasites partially and dose-dependently rescue heat-shock tolerance of PfVps60 knockout parasites [#2]"]}},"prefetch_data":{},"corpus_meta":[{"pmid":"33798247","id":"PMC_33798247","title":"The ESCRT-III machinery participates in the production of extracellular vesicles and protein export during Plasmodium falciparum infection.","date":"2021","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/33798247","citation_count":45,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["PfVPS60"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"42253101","id":"PMC_42253101","title":"Extracellular vesicles participate in proteostasis and heat shock adaptation in Plasmodium falciparum.","date":"2026","source":"Emerging microbes & infections","url":"https://pubmed.ncbi.nlm.nih.gov/42253101","citation_count":0,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["PfVPS60"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":4035,"output_tokens":1079,"usd":0.014145,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":5856,"output_tokens":2215,"usd":0.042327,"stage2_stop_reason":"end_turn"},"total_usd":0.056472,"stage1_batch_id":"msgbatch_011BPjN2BL33F8cJXD1r7V7T","stage2_batch_id":"msgbatch_01Qwg6vP7qxkpL6Gsc7tajsb","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2021,\n      \"finding\": \"PF3D7_1441800 (VPS60/PfVps60) is a component of the P. falciparum ESCRT-III machinery and participates in extracellular vesicle production. Disruption of the PfVps60 gene reduced the number of EVs produced and affected the distribution of other ESCRT-III components. Purified recombinant PfVps60 protein, together with PfVps32 and PfBro1, reconstituted multivesicular body formation and membrane shedding in giant unilamellar vesicles. PfVps60 was confirmed present in EVs purified from parasitized RBC cultures by super-resolution microscopy and dot blot assays.\",\n      \"method\": \"Genetic knockout, in vitro reconstitution with purified recombinant proteins in giant unilamellar vesicles, super-resolution microscopy, dot blot\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — knockout phenotype combined with in vitro reconstitution using purified recombinant proteins, plus direct detection by super-resolution microscopy; multiple orthogonal methods in a single study\",\n      \"pmids\": [\"33798247\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"The P. falciparum ESCRT-III sub-complex is recruited via an alternative pathway involving PfBro1 acting upstream of PfVps32 and PfVps60, differing from canonical eukaryotic ESCRT recruitment.\",\n      \"method\": \"Functional reconstitution with purified recombinant proteins, genetic knockout analysis\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional reconstitution supports pathway order but mechanistic detail of recruitment order inferred from reconstitution data in a single study\",\n      \"pmids\": [\"33798247\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"In P. falciparum 3D7, VPS60 (PF3D7_1441800) knockout impaired EV-mediated export of chaperones PfHsp70-x and PfHsp110 under heat shock conditions, altered protein aggregate dynamics (inducing redistribution of aggregates near the parasitophorous vacuole), reduced induction of cytosolic chaperone PfHsp70-1, and resulted in early loss of parasite viability following heat shock. Supplementation of PfVps60KO parasites with EVs from heat-stressed wild-type parasites partially rescued heat shock tolerance in a dose-dependent manner, linking ESCRT-dependent vesicular trafficking to proteostasis.\",\n      \"method\": \"Genetic knockout (PfVps60KO line), proteomic profiling, fluorescence microscopy (aggregate distribution), viability assays, EV supplementation rescue experiment\",\n      \"journal\": \"Emerging microbes & infections\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean knockout with multiple orthogonal readouts (proteomics, microscopy, viability, rescue) in a single lab study\",\n      \"pmids\": [\"42253101\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Plasmodium falciparum\", \"strain\": \"3D7\"}\n      ]\n    }\n  ],\n  \"current_model\": \"PF3D7_1441800 (VPS60) is a P. falciparum ESCRT-III subunit recruited via PfBro1 that drives extracellular vesicle biogenesis (multivesicular body formation and membrane shedding), is itself packaged into EVs, and mediates EV-dependent export of chaperones (PfHsp70-x, PfHsp110) required for proteostasis and parasite survival under febrile heat-shock conditions.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"PF3D7_1441800 (VPS60) is a component of the Plasmodium falciparum ESCRT-III machinery that drives extracellular vesicle (EV) biogenesis and links vesicular trafficking to parasite proteostasis [#0, #2]. Genetic disruption of VPS60 reduces EV production and perturbs the distribution of other ESCRT-III components, and purified recombinant PfVps60 acts together with PfVps32 and PfBro1 to reconstitute multivesicular body formation and membrane shedding in giant unilamellar vesicles; PfVps60 is itself detected in EVs purified from parasitized red blood cell cultures [#0]. Recruitment of this ESCRT-III sub-complex proceeds through a non-canonical route in which PfBro1 acts upstream of PfVps32 and PfVps60 [#1]. Functionally, VPS60 is required for EV-mediated export of the chaperones PfHsp70-x and PfHsp110 under heat shock, and its loss alters protein aggregate dynamics, dampens induction of cytosolic PfHsp70-1, and causes early loss of parasite viability after heat shock, a defect partially rescued by supplying EVs from heat-stressed wild-type parasites [#2].\",\n  \"teleology\": [\n    {\n      \"year\": 2021,\n      \"claim\": \"Whether PF3D7_1441800 functions in the parasite's ESCRT machinery and contributes to vesicle formation was unknown; establishing this defined its role in EV biogenesis.\",\n      \"evidence\": \"Genetic knockout reducing EV number, in vitro reconstitution of MVB formation and membrane shedding in giant unilamellar vesicles with recombinant PfVps60/PfVps32/PfBro1, super-resolution microscopy and dot blot detection in EVs\",\n      \"pmids\": [\"33798247\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Structural basis of PfVps60 membrane remodeling not resolved\",\n        \"Stoichiometry and assembly geometry of the reconstituted complex not defined\",\n        \"Cargo selectivity of VPS60-dependent EVs not addressed in this study\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"How the ESCRT-III sub-complex including VPS60 is recruited was unclear; ordering PfBro1 upstream of PfVps32 and PfVps60 revealed a non-canonical recruitment pathway.\",\n      \"evidence\": \"Functional reconstitution with purified recombinant proteins combined with genetic knockout analysis\",\n      \"pmids\": [\"33798247\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Recruitment order inferred from reconstitution rather than direct in-cell temporal measurement\",\n        \"Molecular interface between PfBro1 and PfVps60 not mapped\",\n        \"Whether additional adaptors participate in recruitment is unknown\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Whether VPS60-dependent EVs serve a physiological function was open; linking VPS60 loss to failed chaperone export and heat-shock survival established a role in EV-dependent proteostasis.\",\n      \"evidence\": \"PfVps60KO line in P. falciparum 3D7 with proteomic profiling, aggregate-distribution microscopy, viability assays, and dose-dependent EV supplementation rescue\",\n      \"pmids\": [\"42253101\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Molecular mechanism by which chaperones are loaded into VPS60-dependent EVs is unknown\",\n        \"How exported EV chaperones act on recipient cells or the parasite surface is undefined\",\n        \"Only partial rescue achieved, leaving other VPS60-dependent functions unaccounted for\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"3D7\"}]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The structural mechanism of PfVps60-driven membrane remodeling and the basis of chaperone cargo selection into EVs remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No structural model of PfVps60 assembly on membranes\",\n        \"Cargo-sorting mechanism directing PfHsp70-x/PfHsp110 into EVs unknown\",\n        \"Regulation of VPS60 activity under febrile stress not defined\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0032509\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0061512\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:1903561\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0005771\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0036258\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0071364\", \"supporting_discovery_ids\": [2]},\n      {\"term_id\": \"GO:0009408\", \"supporting_discovery_ids\": [2]}\n    ],\n    \"complexes\": [\n      \"ESCRT-III (P. falciparum, with PfVps32 and PfBro1)\"\n    ],\n    \"partners\": [\n      \"PfVps32\",\n      \"PfBro1\",\n      \"PfHsp70-x (EV cargo)\",\n      \"PfHsp110 (EV cargo)\"\n    ],\n    \"other_free_text\": [\n      \"VPS60 loss redistributes protein aggregates near the parasitophorous vacuole and dampens induction of cytosolic PfHsp70-1 [#2]\",\n      \"PfVps60 is itself packaged into extracellular vesicles released from parasitized red blood cells [#0]\",\n      \"EVs from heat-stressed wild-type parasites partially and dose-dependently rescue heat-shock tolerance of PfVps60 knockout parasites [#2]\"\n    ]\n  }\n}","audit_flag":null,"strains":[{"strain":"Pf3D7","gene_id":"PF3D7_1441800","name":null,"product":"vacuolar protein sorting-associated protein 60, putative","uniprot_acc":null,"chromosome":null,"start_pos":null},{"strain":"PbANKA","gene_id":"PBANKA_1305700","name":null,"product":"vacuolar protein sorting-associated protein 60, putative","uniprot_acc":null,"chromosome":null,"start_pos":null},{"strain":"PkH","gene_id":"PKNH_1240500","name":null,"product":"vacuolar protein sorting-associated protein 60, putative","uniprot_acc":null,"chromosome":null,"start_pos":null}],"gene_symbol":"VPS60","evaluation":null}