{"gene":"TG_228750","gene_symbol":"CDPK7","run_date":"2026-09-08T11:06:48","timeline":{"discoveries":[{"year":2013,"finding":"Knock-down of CDPK7 in T. gondii causes pronounced defects in parasite division and major growth deficiency, while being dispensable for motility, egress, and microneme exocytosis, indicating CDPK7 is specifically required for endodyogeny rather than invasion/egress pathways.","method":"Conditional knock-down (protein depletion) with phenotypic analysis of division, motility, egress, and microneme exocytosis","journal":"Cellular microbiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean conditional KD with multiple defined cellular phenotypes in a single lab, several orthogonal readouts","pmids":["24011186"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2013,"finding":"CDPK7 depletion disrupts centrosome integrity and affects the polarity of daughter cell budding and fate of kinetochore-associated proteins, suggesting CDPK7 is required for proper centrosome function at the initiation of endodyogeny.","method":"Conditional knock-down followed by immunofluorescence analysis of centrosomes and kinetochore markers during intracellular replication","journal":"Cellular microbiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KD with defined subcellular phenotypes for centrosome and kinetochore markers, single lab","pmids":["24011186"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2016,"finding":"Complete deletion of CDPK7 by CRISPR/Cas9 confirmed its essentiality for parasite growth in vitro in both type 1 and type 2 genetic backgrounds.","method":"CRISPR/Cas9-mediated complete gene deletion with in vitro growth assay","journal":"Infection and immunity","confidence":"High","confidence_rationale":"Tier 2 / Strong — CRISPR knockout with confirmed essentiality replicated across two genetic backgrounds, corroborating prior conditional KD","pmids":["26755159"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":"RH; ME49"}]},{"year":2021,"finding":"Quantitative phosphoproteomics of TgCDPK7-deficient parasites identified TgRab11a as a substrate of TgCDPK7; phosphorylation of TgRab11a by TgCDPK7 was shown to be critical for parasite intracellular development and protein trafficking.","method":"Quantitative phosphoproteomics of cdpk7-null parasites combined with biochemical and cellular validation of TgRab11a phosphorylation","journal":"PLoS pathogens","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — phosphoproteomics plus biochemical/cellular validation identifying a direct substrate and functional consequence, single lab with multiple orthogonal methods","pmids":["33635921"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2021,"finding":"TgCDPK7 regulates vesicular trafficking and phosphatidylethanolamine (PE) levels in T. gondii, as demonstrated by lipidomic analysis combined with biochemical and cellular studies in cdpk7-deficient parasites.","method":"Lipidomic analysis and biochemical/cellular studies in TgCDPK7-deficient parasites","journal":"PLoS pathogens","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (lipidomics, biochemical, cellular) in a single lab demonstrating pathway involvement","pmids":["33635921"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]}],"current_model":"TgCDPK7 is an essential calcium-dependent protein kinase in Toxoplasma gondii that drives intracellular replication by maintaining centrosome integrity at the onset of endodyogeny, phosphorylating TgRab11a to regulate vesicular trafficking and protein secretion, and controlling phosphatidylethanolamine synthesis, while being dispensable for motility, egress, and microneme exocytosis."},"narrative":{"mechanistic_narrative":"TgCDPK7 is an essential calcium-dependent protein kinase in Toxoplasma gondii that drives intracellular replication by supporting the division cycle rather than the invasion or egress machinery [PMID:24011186]. Its depletion causes pronounced defects in parasite division while leaving motility, egress, and microneme exocytosis intact, marking it as specifically required for endodyogeny [PMID:24011186]. Mechanistically, TgCDPK7 maintains centrosome integrity at the initiation of endodyogeny, and its loss disrupts the polarity of daughter cell budding and the fate of kinetochore-associated proteins [PMID:24011186]. TgCDPK7 phosphorylates TgRab11a, and this phosphorylation is critical for intracellular development and protein trafficking [PMID:33635921], while the kinase also regulates vesicular trafficking and phosphatidylethanolamine levels [PMID:33635921]. Complete gene deletion confirms that TgCDPK7 is essential for growth in both type 1 (RH) and type 2 (ME49) backgrounds [PMID:26755159].","teleology":[{"year":2013,"claim":"Establishing which stage of the lytic cycle requires TgCDPK7 answered whether this kinase acts in invasion/egress or in replication, localizing its function to intracellular division.","evidence":"Conditional knock-down with phenotypic readouts for division, motility, egress, and microneme exocytosis in T. gondii","pmids":["24011186"],"confidence":"Medium","gaps":["Does not identify direct substrates or molecular partners","Mechanism linking the kinase to the division defect unresolved"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2013,"claim":"Pinpointing the subcellular structures perturbed on depletion showed TgCDPK7 is required for centrosome function at the onset of endodyogeny, explaining the division defect.","evidence":"Conditional knock-down with immunofluorescence of centrosome and kinetochore markers during replication","pmids":["24011186"],"confidence":"Medium","gaps":["No molecular target at the centrosome identified","How the kinase controls budding polarity mechanistically is unknown"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2016,"claim":"Complete gene deletion resolved whether the growth requirement was strain-restricted, confirming essentiality across type 1 and type 2 backgrounds.","evidence":"CRISPR/Cas9 knockout with in vitro growth assay in RH and ME49","pmids":["26755159"],"confidence":"High","gaps":["Does not address the molecular basis of essentiality","No in vivo/virulence assessment"],"attribution":[{"species":"Toxoplasma gondii","strain":"RH; ME49"}]},{"year":2021,"claim":"Identifying a direct substrate answered how the kinase exerts its function, linking TgCDPK7 to TgRab11a phosphorylation as a driver of intracellular development and trafficking.","evidence":"Quantitative phosphoproteomics of cdpk7-null parasites with biochemical and cellular validation of TgRab11a phosphorylation","pmids":["33635921"],"confidence":"High","gaps":["Phosphosite(s) on TgRab11a and their functional impact not fully mapped","Whether TgRab11a phosphorylation accounts for the centrosome defect is unresolved"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2021,"claim":"Coupling lipidomics to the null mutant connected the kinase to membrane biogenesis, showing TgCDPK7 regulates vesicular trafficking and phosphatidylethanolamine levels.","evidence":"Lipidomic analysis with biochemical and cellular studies in TgCDPK7-deficient parasites","pmids":["33635921"],"confidence":"Medium","gaps":["Direct enzymatic link between the kinase and PE synthesis enzymes not established","Relationship between the trafficking defect and altered PE unresolved"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":null,"claim":"How the calcium signal is sensed and how substrate phosphorylation is coordinated across centrosome integrity, Rab11a-dependent trafficking, and lipid metabolism remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model of the kinase","Upstream calcium-signaling context uncharacterized","Full substrate repertoire beyond TgRab11a not defined"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]}],"mechanism_profile":{"molecular_activity":[],"localization":[],"pathway":[],"complexes":[],"partners":["TGRAB11A"],"other_free_text":["Essential for parasite growth in both type 1 (RH) and type 2 (ME49) backgrounds [#2].","Required for endodyogeny/intracellular replication but dispensable for motility, egress, and microneme exocytosis [#0].","Maintains centrosome integrity and daughter-cell budding polarity at the onset of endodyogeny [#1]."]},"strain_conflicts":[]},"prefetch_data":{},"corpus_meta":[{"pmid":"24011186","id":"PMC_24011186","title":"The Toxoplasma gondii calcium-dependent protein kinase 7 is involved in early steps of parasite division and is crucial for parasite survival.","date":"2013","source":"Cellular microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/24011186","citation_count":76,"is_preprint":false},{"pmid":"26755159","id":"PMC_26755159","title":"Analysis of Noncanonical Calcium-Dependent Protein Kinases in Toxoplasma gondii by Targeted Gene Deletion Using CRISPR/Cas9.","date":"2016","source":"Infection and immunity","url":"https://pubmed.ncbi.nlm.nih.gov/26755159","citation_count":56,"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":"34336254","id":"PMC_34336254","title":"Immunoinformatic Analysis of Calcium-Dependent Protein Kinase 7 (CDPK7) Showed Potential Targets for Toxoplasma gondii Vaccine.","date":"2021","source":"Journal of parasitology research","url":"https://pubmed.ncbi.nlm.nih.gov/34336254","citation_count":5,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":4194,"output_tokens":1273,"usd":0.015838,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":6294,"output_tokens":2281,"usd":0.044248,"stage2_stop_reason":"end_turn"},"total_usd":0.060086,"stage1_batch_id":"msgbatch_01WokvBYTB6JLnYrpD4rXnyV","stage2_batch_id":"msgbatch_01M7hsQDvz6AqVC9DLVjQzzc","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2013,\n      \"finding\": \"Knock-down of CDPK7 in T. gondii causes pronounced defects in parasite division and major growth deficiency, while being dispensable for motility, egress, and microneme exocytosis, indicating CDPK7 is specifically required for endodyogeny rather than invasion/egress pathways.\",\n      \"method\": \"Conditional knock-down (protein depletion) with phenotypic analysis of division, motility, egress, and microneme exocytosis\",\n      \"journal\": \"Cellular microbiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean conditional KD with multiple defined cellular phenotypes in a single lab, several orthogonal readouts\",\n      \"pmids\": [\"24011186\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"CDPK7 depletion disrupts centrosome integrity and affects the polarity of daughter cell budding and fate of kinetochore-associated proteins, suggesting CDPK7 is required for proper centrosome function at the initiation of endodyogeny.\",\n      \"method\": \"Conditional knock-down followed by immunofluorescence analysis of centrosomes and kinetochore markers during intracellular replication\",\n      \"journal\": \"Cellular microbiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KD with defined subcellular phenotypes for centrosome and kinetochore markers, single lab\",\n      \"pmids\": [\"24011186\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Complete deletion of CDPK7 by CRISPR/Cas9 confirmed its essentiality for parasite growth in vitro in both type 1 and type 2 genetic backgrounds.\",\n      \"method\": \"CRISPR/Cas9-mediated complete gene deletion with in vitro growth assay\",\n      \"journal\": \"Infection and immunity\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — CRISPR knockout with confirmed essentiality replicated across two genetic backgrounds, corroborating prior conditional KD\",\n      \"pmids\": [\"26755159\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"RH; ME49\"}]\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Quantitative phosphoproteomics of TgCDPK7-deficient parasites identified TgRab11a as a substrate of TgCDPK7; phosphorylation of TgRab11a by TgCDPK7 was shown to be critical for parasite intracellular development and protein trafficking.\",\n      \"method\": \"Quantitative phosphoproteomics of cdpk7-null parasites combined with biochemical and cellular validation of TgRab11a phosphorylation\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — phosphoproteomics plus biochemical/cellular validation identifying a direct substrate and functional consequence, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"33635921\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"TgCDPK7 regulates vesicular trafficking and phosphatidylethanolamine (PE) levels in T. gondii, as demonstrated by lipidomic analysis combined with biochemical and cellular studies in cdpk7-deficient parasites.\",\n      \"method\": \"Lipidomic analysis and biochemical/cellular studies in TgCDPK7-deficient parasites\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (lipidomics, biochemical, cellular) in a single lab demonstrating pathway involvement\",\n      \"pmids\": [\"33635921\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    }\n  ],\n  \"current_model\": \"TgCDPK7 is an essential calcium-dependent protein kinase in Toxoplasma gondii that drives intracellular replication by maintaining centrosome integrity at the onset of endodyogeny, phosphorylating TgRab11a to regulate vesicular trafficking and protein secretion, and controlling phosphatidylethanolamine synthesis, while being dispensable for motility, egress, and microneme exocytosis.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"TgCDPK7 is an essential calcium-dependent protein kinase in Toxoplasma gondii that drives intracellular replication by supporting the division cycle rather than the invasion or egress machinery [#0]. Its depletion causes pronounced defects in parasite division while leaving motility, egress, and microneme exocytosis intact, marking it as specifically required for endodyogeny [#0]. Mechanistically, TgCDPK7 maintains centrosome integrity at the initiation of endodyogeny, and its loss disrupts the polarity of daughter cell budding and the fate of kinetochore-associated proteins [#1]. TgCDPK7 phosphorylates TgRab11a, and this phosphorylation is critical for intracellular development and protein trafficking [#3], while the kinase also regulates vesicular trafficking and phosphatidylethanolamine levels [#4]. Complete gene deletion confirms that TgCDPK7 is essential for growth in both type 1 (RH) and type 2 (ME49) backgrounds [#2].\",\n  \"teleology\": [\n    {\n      \"year\": 2013,\n      \"claim\": \"Establishing which stage of the lytic cycle requires TgCDPK7 answered whether this kinase acts in invasion/egress or in replication, localizing its function to intracellular division.\",\n      \"evidence\": \"Conditional knock-down with phenotypic readouts for division, motility, egress, and microneme exocytosis in T. gondii\",\n      \"pmids\": [\"24011186\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Does not identify direct substrates or molecular partners\", \"Mechanism linking the kinase to the division defect unresolved\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Pinpointing the subcellular structures perturbed on depletion showed TgCDPK7 is required for centrosome function at the onset of endodyogeny, explaining the division defect.\",\n      \"evidence\": \"Conditional knock-down with immunofluorescence of centrosome and kinetochore markers during replication\",\n      \"pmids\": [\"24011186\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No molecular target at the centrosome identified\", \"How the kinase controls budding polarity mechanistically is unknown\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Complete gene deletion resolved whether the growth requirement was strain-restricted, confirming essentiality across type 1 and type 2 backgrounds.\",\n      \"evidence\": \"CRISPR/Cas9 knockout with in vitro growth assay in RH and ME49\",\n      \"pmids\": [\"26755159\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Does not address the molecular basis of essentiality\", \"No in vivo/virulence assessment\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"RH; ME49\"}]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Identifying a direct substrate answered how the kinase exerts its function, linking TgCDPK7 to TgRab11a phosphorylation as a driver of intracellular development and trafficking.\",\n      \"evidence\": \"Quantitative phosphoproteomics of cdpk7-null parasites with biochemical and cellular validation of TgRab11a phosphorylation\",\n      \"pmids\": [\"33635921\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Phosphosite(s) on TgRab11a and their functional impact not fully mapped\", \"Whether TgRab11a phosphorylation accounts for the centrosome defect is unresolved\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Coupling lipidomics to the null mutant connected the kinase to membrane biogenesis, showing TgCDPK7 regulates vesicular trafficking and phosphatidylethanolamine levels.\",\n      \"evidence\": \"Lipidomic analysis with biochemical and cellular studies in TgCDPK7-deficient parasites\",\n      \"pmids\": [\"33635921\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct enzymatic link between the kinase and PE synthesis enzymes not established\", \"Relationship between the trafficking defect and altered PE unresolved\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the calcium signal is sensed and how substrate phosphorylation is coordinated across centrosome integrity, Rab11a-dependent trafficking, and lipid metabolism remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model of the kinase\", \"Upstream calcium-signaling context uncharacterized\", \"Full substrate repertoire beyond TgRab11a not defined\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0004683\", \"supporting_discovery_ids\": [3], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005813\", \"supporting_discovery_ids\": [1], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0016192\", \"supporting_discovery_ids\": [3, 4], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]},\n      {\"term_id\": \"GO:0006646\", \"supporting_discovery_ids\": [4], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"TgRab11a\"],\n    \"other_free_text\": [\n      \"Essential for parasite growth in both type 1 (RH) and type 2 (ME49) backgrounds [#2].\",\n      \"Required for endodyogeny/intracellular replication but dispensable for motility, egress, and microneme exocytosis [#0].\",\n      \"Maintains centrosome integrity and daughter-cell budding polarity at the onset of endodyogeny [#1].\"\n    ]\n  },\n  \"strain_conflicts\": []\n}","audit_flag":null,"strains":[{"strain":"ME49","gene_id":"TGME49_228750","name":"CDPK7","product":"calcium dependent protein kinase CDPK7","uniprot_acc":"S8G6L4","chromosome":"TGME49_chrX","start_pos":159878},{"strain":"GT1","gene_id":"TGGT1_228750","name":null,"product":"calcium dependent protein kinase CDPK7","uniprot_acc":null,"chromosome":"TGGT1_chrX","start_pos":72512},{"strain":"RH88","gene_id":"TGRH88_047930","name":null,"product":"calcium dependent protein kinase CDPK7","uniprot_acc":null,"chromosome":"CM023091","start_pos":7398545},{"strain":"VEG","gene_id":"TGVEG_228750","name":null,"product":"calcium dependent protein kinase CDPK7","uniprot_acc":"V4YXF7","chromosome":"TGVEG_chrX","start_pos":51829}]}