{"gene":"PF3D7_0307300","run_date":"2026-09-07T10:55:51","timeline":{"discoveries":[{"year":2023,"finding":"Plasmodium EB1 is an orthologue distinct from canonical eukaryotic EB1: in vitro and in vivo assays demonstrate it lacks microtubule plus-end tracking but possesses microtubule-lattice binding affinity, contributed by both its CH domain and linker region.","method":"In vitro microtubule binding assays and in vivo live-cell imaging; domain mutagenesis/analysis","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution plus in vivo assays with domain-level dissection, single lab but two orthogonal methods","pmids":["37208365"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2023,"finding":"EB1 localizes to the nucleoplasm of male gametocytes and, during gametogenesis, decorates the full length of spindle microtubules (not just plus-ends), regulating spindle structure and enabling lateral kinetochore-spindle attachment throughout endomitosis in an EB1-dependent manner.","method":"Fluorescence microscopy (live-cell and fixed), EB1-deficient parasite lines","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Moderate — direct localization with functional KO phenotype and orthogonal imaging methods, single lab","pmids":["37208365"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2023,"finding":"Deletion of EB1 in P. falciparum produces male gametocytes that develop into anucleated male gametes (severe defect in nuclear segregation/partitioning into emerging microgametes), revealing an essential male-specific fertility function; asexual proliferation and gametocytogenesis are unaffected.","method":"Gene deletion (KO), genetic crosses with male- and female-sterile parasite lines, microscopy","journal":"mBio","confidence":"High","confidence_rationale":"Tier 2 / Strong — independently corroborated by two separate labs (PMID 37208365 and 37535401) using gene deletion with specific phenotypic readouts","pmids":["37535401","37208365"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":"NF54"}]},{"year":2023,"finding":"EB1 is identified as a physical interactor of the aurora-related kinase ARK2 in Plasmodium berghei sexual stages; interactomic (mass spectrometry) and co-localization studies place EB1 at spindle microtubules in the vicinity of kinetochores during both mitosis and meiosis.","method":"Mass spectrometry-based interactomics (co-immunoprecipitation/pulldown), super-resolution and live-cell fluorescence microscopy","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal interactomics plus co-localization, single lab, two orthogonal methods","pmids":["37704606"],"is_preprint":false,"attribution":[{"species":"Plasmodium berghei","strain":""}]},{"year":2023,"finding":"Gene function studies show that ARK2 and EB1 have complementary (non-redundant) roles in driving endomitotic division and parasite transmission through the mosquito in P. berghei sexual stages.","method":"Genetic epistasis / gene deletion of ARK2 and EB1, mosquito transmission assays","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis with transmission phenotype, single lab, two gene KOs tested","pmids":["37704606"],"is_preprint":false,"attribution":[{"species":"Plasmodium berghei","strain":""}]},{"year":2022,"finding":"In P. falciparum gametocytes, EB1 (end-binding protein-1) is a component of classical mitotic machinery recruited to a non-mitotic MTOC embedded in the nuclear membrane, where it participates in nuclear microtubule assembly/disassembly processes that drive nuclear elongation and chromatin organization during gametocyte development.","method":"3D electron microscopy, fluorescence microscopy, microtubule depolymerization (trifluralin treatment)","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — 3D EM plus pharmacological perturbation, direct localization with functional consequence, single lab","pmids":["36030238"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2023,"finding":"PfEB1 is hypophosphorylated in Pfcdpk4-deleted gametocytes, placing it downstream of Ca2+-dependent protein kinase 4 (CDPK4) in a signaling pathway regulating microtubule dynamics during male gametogenesis.","method":"Comparative phospho-proteome analysis of Pfcdpk4 knockout gametocytes","journal":"mBio","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single phosphoproteomics dataset, no direct functional validation of the phosphorylation event on EB1 itself","pmids":["37535401"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]}],"current_model":"Plasmodium falciparum EB1 (PF3D7_0307300) is a divergent end-binding protein that, unlike canonical eukaryotic EB1, lacks microtubule plus-end tracking and instead binds along the full microtubule lattice via its CH domain and linker region; it localizes to spindle microtubules and the nucleoplasm during male gametogenesis, where it is essential for lateral kinetochore–spindle attachment, nuclear segregation, and partitioning of nuclei into male gametes, acts as a physical interactor of aurora-related kinase ARK2 at the spindle, and is regulated upstream by CDPK4-dependent phosphorylation."},"narrative":{"mechanistic_narrative":"EB1 (PF3D7_0307300) is a divergent Plasmodium end-binding protein that governs spindle-based nuclear division during male gametogenesis [PMID:37208365]. Unlike canonical eukaryotic EB1, it does not track microtubule plus-ends; instead it binds along the full microtubule lattice through contributions from both its CH domain and linker region [PMID:37208365]. In P. falciparum male gametocytes it resides in the nucleoplasm and decorates the entire length of spindle microtubules during endomitosis, where it shapes spindle structure and enables lateral kinetochore–spindle attachment [PMID:37208365]. Consistent with this role, EB1 deletion yields male gametocytes that produce anucleated microgametes owing to failed nuclear segregation/partitioning, defining an essential, male-specific fertility function that leaves asexual proliferation and gametocytogenesis intact [PMID:37535401, PMID:37208365]. EB1 is recruited to a nuclear-membrane-embedded MTOC and participates in nuclear microtubule assembly/disassembly driving nuclear elongation and chromatin organization [PMID:36030238]. In P. berghei sexual stages, EB1 physically associates with the aurora-related kinase ARK2 at spindle microtubules near kinetochores, and the two proteins play complementary, non-redundant roles in endomitotic division and mosquito transmission [PMID:37704606]. Upstream, EB1 is hypophosphorylated in CDPK4-null gametocytes, placing it downstream of CDPK4-dependent signaling [PMID:37535401].","teleology":[{"year":2022,"claim":"Before its molecular role was defined, it was unknown how mitotic machinery organized the unusual nuclear division of Plasmodium gametocytes; this established EB1 as a component recruited to a nuclear-membrane MTOC driving nuclear microtubule dynamics.","evidence":"3D electron microscopy, fluorescence microscopy and trifluralin microtubule depolymerization in P. falciparum gametocytes","pmids":["36030238"],"confidence":"Medium","gaps":["Does not resolve which microtubule-binding mode EB1 uses","Does not distinguish EB1's contribution from other MTOC components"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2023,"claim":"It was unknown whether Plasmodium EB1 behaves like canonical plus-end-tracking EB1; in vitro reconstitution with domain dissection showed it instead binds the full microtubule lattice via its CH domain and linker, redefining it as a divergent orthologue.","evidence":"In vitro microtubule binding assays, in vivo live-cell imaging and domain mutagenesis in P. falciparum","pmids":["37208365"],"confidence":"High","gaps":["Structural basis of lattice binding not resolved","Functional consequence of removing individual binding determinants in vivo not fully mapped"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2023,"claim":"Whether EB1's lattice binding served a specific division function was open; localization and knockout imaging established that EB1 coats spindle microtubules and is required for lateral kinetochore–spindle attachment during endomitosis.","evidence":"Live-cell and fixed fluorescence microscopy in EB1-deficient P. falciparum lines","pmids":["37208365"],"confidence":"High","gaps":["Molecular link between lattice binding and kinetochore attachment not defined","Does not identify the kinetochore-side partner"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2023,"claim":"The biological importance of EB1 was unresolved; gene deletion demonstrated an essential, stage-specific role, as EB1-null males form anucleated gametes while asexual growth and gametocytogenesis are unaffected.","evidence":"Gene deletion, genetic crosses with male/female-sterile lines and microscopy in P. falciparum NF54, corroborated by two labs","pmids":["37535401","37208365"],"confidence":"High","gaps":["Mechanism linking segregation failure to anucleate gametes not fully dissected","Female-side contribution not characterized"],"attribution":[{"species":"Plasmodium falciparum","strain":"NF54"}]},{"year":2023,"claim":"How EB1 is coordinated with mitotic kinases was unknown; interactomics and genetics in P. berghei placed EB1 as a physical partner of ARK2 at spindle microtubules with complementary, non-redundant roles in division and transmission.","evidence":"Mass spectrometry interactomics, super-resolution/live-cell imaging and gene deletion with mosquito transmission assays in P. berghei","pmids":["37704606"],"confidence":"Medium","gaps":["Direct binding interface between EB1 and ARK2 not mapped","Whether ARK2 regulates EB1 activity or vice versa is unresolved","Shown in P. berghei; direct interaction not verified in P. falciparum"],"attribution":[{"species":"Plasmodium berghei","strain":""}]},{"year":2023,"claim":"The upstream regulatory input to EB1 was unknown; comparative phosphoproteomics showed EB1 is hypophosphorylated in CDPK4-null gametocytes, placing it downstream of CDPK4 signaling.","evidence":"Comparative phospho-proteome analysis of Pfcdpk4 knockout gametocytes","pmids":["37535401"],"confidence":"Low","gaps":["Single phosphoproteomics dataset without functional validation of the phosphosite on EB1","Whether phosphorylation is direct or indirect not established","Functional effect of the modification on EB1 activity unknown"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":null,"claim":"How EB1's lattice-binding activity mechanistically drives lateral kinetochore–spindle attachment, and how CDPK4 phosphorylation and ARK2 association converge to control this during male gametogenesis, remain unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["No structural model of EB1 on the lattice or at kinetochores","Causal chain from CDPK4 phosphorylation to EB1 function not established","EB1–ARK2 interaction interface and regulatory hierarchy undefined"],"attribution":[]}],"mechanism_profile":{"molecular_activity":[],"localization":[],"pathway":[],"complexes":[],"partners":["ARK2 (AURORA-RELATED KINASE; PHYSICAL INTERACTOR IN P. BERGHEI SEXUAL STAGES) [#3]","CDPK4 (CA2+-DEPENDENT PROTEIN KINASE 4; UPSTREAM REGULATOR, EB1 HYPOPHOSPHORYLATED IN CDPK4-NULL GAMETOCYTES) [#6]"],"other_free_text":["Divergent end-binding protein that binds the full microtubule lattice rather than tracking plus-ends; binding contributed by both CH domain and linker region [#0].","Essential for male-specific fertility: deletion produces anucleated microgametes via failed nuclear segregation, while asexual proliferation and gametocytogenesis are unaffected [#2].","Recruited to a nuclear-membrane-embedded MTOC and participates in nuclear microtubule assembly/disassembly driving nuclear elongation and chromatin organization [#5]."]}},"prefetch_data":{},"corpus_meta":[{"pmid":"36030238","id":"PMC_36030238","title":"Repurposing the mitotic machinery to drive cellular elongation and chromatin reorganisation in Plasmodium falciparum gametocytes.","date":"2022","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/36030238","citation_count":32,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["end-binding protein 1"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"37704606","id":"PMC_37704606","title":"Plasmodium ARK2 and EB1 drive unconventional spindle dynamics, during chromosome segregation in sexual transmission stages.","date":"2023","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/37704606","citation_count":29,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["EB1"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"37208365","id":"PMC_37208365","title":"EB1 decoration of microtubule lattice facilitates spindle-kinetochore lateral attachment in Plasmodium male gametogenesis.","date":"2023","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/37208365","citation_count":16,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["EB1"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"37535401","id":"PMC_37535401","title":"Plasmodium microtubule-binding protein EB1 is critical for partitioning of nuclei in male gametogenesis.","date":"2023","source":"mBio","url":"https://pubmed.ncbi.nlm.nih.gov/37535401","citation_count":5,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["EB1","end-binding protein 1","PfEB1"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"36798191","id":"PMC_36798191","title":"Plasmodium ARK2-EB1 axis drives the unconventional spindle dynamics, scaffold formation and chromosome segregation of sexual transmission stages.","date":"2023","source":"Research square","url":"https://pubmed.ncbi.nlm.nih.gov/36798191","citation_count":2,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["EB1"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"36778504","id":"PMC_36778504","title":"Plasmodium ARK2-EB1 axis drives the unconventional spindle dynamics, scaffold formation and chromosome segregation of sexual transmission stages.","date":"2023","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/36778504","citation_count":1,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["EB1"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":6148,"output_tokens":1794,"usd":0.022677,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":6919,"output_tokens":3067,"usd":0.055635,"stage2_stop_reason":"end_turn"},"total_usd":0.078312,"stage1_batch_id":"msgbatch_01DbiymDmubkPpYvdbAu5a7V","stage2_batch_id":"msgbatch_012TbrWvMRH129T7PQ4SmvrJ","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2023,\n      \"finding\": \"Plasmodium EB1 is an orthologue distinct from canonical eukaryotic EB1: in vitro and in vivo assays demonstrate it lacks microtubule plus-end tracking but possesses microtubule-lattice binding affinity, contributed by both its CH domain and linker region.\",\n      \"method\": \"In vitro microtubule binding assays and in vivo live-cell imaging; domain mutagenesis/analysis\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution plus in vivo assays with domain-level dissection, single lab but two orthogonal methods\",\n      \"pmids\": [\"37208365\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"EB1 localizes to the nucleoplasm of male gametocytes and, during gametogenesis, decorates the full length of spindle microtubules (not just plus-ends), regulating spindle structure and enabling lateral kinetochore-spindle attachment throughout endomitosis in an EB1-dependent manner.\",\n      \"method\": \"Fluorescence microscopy (live-cell and fixed), EB1-deficient parasite lines\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct localization with functional KO phenotype and orthogonal imaging methods, single lab\",\n      \"pmids\": [\"37208365\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Deletion of EB1 in P. falciparum produces male gametocytes that develop into anucleated male gametes (severe defect in nuclear segregation/partitioning into emerging microgametes), revealing an essential male-specific fertility function; asexual proliferation and gametocytogenesis are unaffected.\",\n      \"method\": \"Gene deletion (KO), genetic crosses with male- and female-sterile parasite lines, microscopy\",\n      \"journal\": \"mBio\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — independently corroborated by two separate labs (PMID 37208365 and 37535401) using gene deletion with specific phenotypic readouts\",\n      \"pmids\": [\"37535401\", \"37208365\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"NF54\"}]\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"EB1 is identified as a physical interactor of the aurora-related kinase ARK2 in Plasmodium berghei sexual stages; interactomic (mass spectrometry) and co-localization studies place EB1 at spindle microtubules in the vicinity of kinetochores during both mitosis and meiosis.\",\n      \"method\": \"Mass spectrometry-based interactomics (co-immunoprecipitation/pulldown), super-resolution and live-cell fluorescence microscopy\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal interactomics plus co-localization, single lab, two orthogonal methods\",\n      \"pmids\": [\"37704606\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium berghei\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Gene function studies show that ARK2 and EB1 have complementary (non-redundant) roles in driving endomitotic division and parasite transmission through the mosquito in P. berghei sexual stages.\",\n      \"method\": \"Genetic epistasis / gene deletion of ARK2 and EB1, mosquito transmission assays\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis with transmission phenotype, single lab, two gene KOs tested\",\n      \"pmids\": [\"37704606\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium berghei\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"In P. falciparum gametocytes, EB1 (end-binding protein-1) is a component of classical mitotic machinery recruited to a non-mitotic MTOC embedded in the nuclear membrane, where it participates in nuclear microtubule assembly/disassembly processes that drive nuclear elongation and chromatin organization during gametocyte development.\",\n      \"method\": \"3D electron microscopy, fluorescence microscopy, microtubule depolymerization (trifluralin treatment)\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — 3D EM plus pharmacological perturbation, direct localization with functional consequence, single lab\",\n      \"pmids\": [\"36030238\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"PfEB1 is hypophosphorylated in Pfcdpk4-deleted gametocytes, placing it downstream of Ca2+-dependent protein kinase 4 (CDPK4) in a signaling pathway regulating microtubule dynamics during male gametogenesis.\",\n      \"method\": \"Comparative phospho-proteome analysis of Pfcdpk4 knockout gametocytes\",\n      \"journal\": \"mBio\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single phosphoproteomics dataset, no direct functional validation of the phosphorylation event on EB1 itself\",\n      \"pmids\": [\"37535401\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    }\n  ],\n  \"current_model\": \"Plasmodium falciparum EB1 (PF3D7_0307300) is a divergent end-binding protein that, unlike canonical eukaryotic EB1, lacks microtubule plus-end tracking and instead binds along the full microtubule lattice via its CH domain and linker region; it localizes to spindle microtubules and the nucleoplasm during male gametogenesis, where it is essential for lateral kinetochore–spindle attachment, nuclear segregation, and partitioning of nuclei into male gametes, acts as a physical interactor of aurora-related kinase ARK2 at the spindle, and is regulated upstream by CDPK4-dependent phosphorylation.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"EB1 (PF3D7_0307300) is a divergent Plasmodium end-binding protein that governs spindle-based nuclear division during male gametogenesis [#0, #1]. Unlike canonical eukaryotic EB1, it does not track microtubule plus-ends; instead it binds along the full microtubule lattice through contributions from both its CH domain and linker region [#0]. In P. falciparum male gametocytes it resides in the nucleoplasm and decorates the entire length of spindle microtubules during endomitosis, where it shapes spindle structure and enables lateral kinetochore–spindle attachment [#1]. Consistent with this role, EB1 deletion yields male gametocytes that produce anucleated microgametes owing to failed nuclear segregation/partitioning, defining an essential, male-specific fertility function that leaves asexual proliferation and gametocytogenesis intact [#2]. EB1 is recruited to a nuclear-membrane-embedded MTOC and participates in nuclear microtubule assembly/disassembly driving nuclear elongation and chromatin organization [#5]. In P. berghei sexual stages, EB1 physically associates with the aurora-related kinase ARK2 at spindle microtubules near kinetochores, and the two proteins play complementary, non-redundant roles in endomitotic division and mosquito transmission [#3, #4]. Upstream, EB1 is hypophosphorylated in CDPK4-null gametocytes, placing it downstream of CDPK4-dependent signaling [#6].\",\n  \"teleology\": [\n    {\n      \"year\": 2022,\n      \"claim\": \"Before its molecular role was defined, it was unknown how mitotic machinery organized the unusual nuclear division of Plasmodium gametocytes; this established EB1 as a component recruited to a nuclear-membrane MTOC driving nuclear microtubule dynamics.\",\n      \"evidence\": \"3D electron microscopy, fluorescence microscopy and trifluralin microtubule depolymerization in P. falciparum gametocytes\",\n      \"pmids\": [\"36030238\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Does not resolve which microtubule-binding mode EB1 uses\", \"Does not distinguish EB1's contribution from other MTOC components\"],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"It was unknown whether Plasmodium EB1 behaves like canonical plus-end-tracking EB1; in vitro reconstitution with domain dissection showed it instead binds the full microtubule lattice via its CH domain and linker, redefining it as a divergent orthologue.\",\n      \"evidence\": \"In vitro microtubule binding assays, in vivo live-cell imaging and domain mutagenesis in P. falciparum\",\n      \"pmids\": [\"37208365\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Structural basis of lattice binding not resolved\", \"Functional consequence of removing individual binding determinants in vivo not fully mapped\"],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Whether EB1's lattice binding served a specific division function was open; localization and knockout imaging established that EB1 coats spindle microtubules and is required for lateral kinetochore–spindle attachment during endomitosis.\",\n      \"evidence\": \"Live-cell and fixed fluorescence microscopy in EB1-deficient P. falciparum lines\",\n      \"pmids\": [\"37208365\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Molecular link between lattice binding and kinetochore attachment not defined\", \"Does not identify the kinetochore-side partner\"],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"The biological importance of EB1 was unresolved; gene deletion demonstrated an essential, stage-specific role, as EB1-null males form anucleated gametes while asexual growth and gametocytogenesis are unaffected.\",\n      \"evidence\": \"Gene deletion, genetic crosses with male/female-sterile lines and microscopy in P. falciparum NF54, corroborated by two labs\",\n      \"pmids\": [\"37535401\", \"37208365\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Mechanism linking segregation failure to anucleate gametes not fully dissected\", \"Female-side contribution not characterized\"],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"NF54\"}]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"How EB1 is coordinated with mitotic kinases was unknown; interactomics and genetics in P. berghei placed EB1 as a physical partner of ARK2 at spindle microtubules with complementary, non-redundant roles in division and transmission.\",\n      \"evidence\": \"Mass spectrometry interactomics, super-resolution/live-cell imaging and gene deletion with mosquito transmission assays in P. berghei\",\n      \"pmids\": [\"37704606\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Direct binding interface between EB1 and ARK2 not mapped\", \"Whether ARK2 regulates EB1 activity or vice versa is unresolved\", \"Shown in P. berghei; direct interaction not verified in P. falciparum\"],\n      \"attribution\": [{\"species\": \"Plasmodium berghei\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"The upstream regulatory input to EB1 was unknown; comparative phosphoproteomics showed EB1 is hypophosphorylated in CDPK4-null gametocytes, placing it downstream of CDPK4 signaling.\",\n      \"evidence\": \"Comparative phospho-proteome analysis of Pfcdpk4 knockout gametocytes\",\n      \"pmids\": [\"37535401\"],\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Single phosphoproteomics dataset without functional validation of the phosphosite on EB1\", \"Whether phosphorylation is direct or indirect not established\", \"Functional effect of the modification on EB1 activity unknown\"],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How EB1's lattice-binding activity mechanistically drives lateral kinetochore–spindle attachment, and how CDPK4 phosphorylation and ARK2 association converge to control this during male gametogenesis, remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"No structural model of EB1 on the lattice or at kinetochores\", \"Causal chain from CDPK4 phosphorylation to EB1 function not established\", \"EB1–ARK2 interaction interface and regulatory hierarchy undefined\"],\n      \"attribution\": []\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008017\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005819\", \"supporting_discovery_ids\": [1, 3, 5]},\n      {\"term_id\": \"GO:0005654\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0000278\", \"supporting_discovery_ids\": [4, 5]},\n      {\"term_id\": \"GO:0007059\", \"supporting_discovery_ids\": [1, 2]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"ARK2 (aurora-related kinase; physical interactor in P. berghei sexual stages) [#3]\",\n      \"CDPK4 (Ca2+-dependent protein kinase 4; upstream regulator, EB1 hypophosphorylated in CDPK4-null gametocytes) [#6]\"\n    ],\n    \"other_free_text\": [\n      \"Divergent end-binding protein that binds the full microtubule lattice rather than tracking plus-ends; binding contributed by both CH domain and linker region [#0].\",\n      \"Essential for male-specific fertility: deletion produces anucleated microgametes via failed nuclear segregation, while asexual proliferation and gametocytogenesis are unaffected [#2].\",\n      \"Recruited to a nuclear-membrane-embedded MTOC and participates in nuclear microtubule assembly/disassembly driving nuclear elongation and chromatin organization [#5].\"\n    ]\n  }\n}","audit_flag":null,"strains":[{"strain":"Pf3D7","gene_id":"PF3D7_0307300","name":null,"product":"end-binding protein 1","uniprot_acc":null,"chromosome":null,"start_pos":null},{"strain":"PbANKA","gene_id":"PBANKA_0405600","name":null,"product":"microtubule-associated protein RP/EB family, putative","uniprot_acc":null,"chromosome":null,"start_pos":null},{"strain":"PkH","gene_id":"PKNH_0835400","name":null,"product":"end-binding protein 1, putative","uniprot_acc":null,"chromosome":null,"start_pos":null}],"gene_symbol":"EB1","evaluation":null}