{"gene":"KLHL25","run_date":"2026-04-28T18:30:27","timeline":{"discoveries":[{"year":2012,"finding":"KLHL25 forms a complex with CUL3 (KLHL25-CUL3) that acts as an E3 ubiquitin ligase targeting hypophosphorylated 4E-BP1 for ubiquitination and degradation, thereby providing homeostatic control of eIF4E activity.","method":"Biochemical identification of E3 ubiquitin ligase complex; knockdown experiments in cells showing 4E-BP1 degradation dependent on phosphorylation status","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods (RNAi knockdown, co-immunoprecipitation, ubiquitination assays) in a highly-cited foundational study","pmids":["22578813"],"is_preprint":false},{"year":2016,"finding":"KLHL25 serves as the adaptor protein for the CUL3-RING ubiquitin ligase complex that binds ACLY (ATP-citrate lyase), mediating its ubiquitination and proteasomal degradation to inhibit lipid synthesis.","method":"Co-immunoprecipitation showing CUL3-KLHL25 interaction with ACLY; ubiquitination assays; xenograft tumor growth assays with loss-of-function","journal":"Genes & development","confidence":"High","confidence_rationale":"Tier 2 — reciprocal Co-IP, ubiquitination assay, and in vivo tumor model with functional readout in a well-cited study","pmids":["27664236"],"is_preprint":false},{"year":2021,"finding":"TGFβ1 stimulation during iTreg differentiation induces CUL3-KLHL25-mediated ubiquitination and degradation of ACLY, which reduces malonyl-CoA levels, relieving inhibition of CPT1 and thereby shifting fatty acid metabolism from synthesis to oxidation to support iTreg differentiation.","method":"Biochemical ubiquitination assays, metabolic profiling (malonyl-CoA measurement), iTreg differentiation assays with KLHL25 knockdown/overexpression, CPT1 activity measurement","journal":"eLife","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods (ubiquitination assay, metabolomics, functional differentiation assay) in a single rigorous study","pmids":["34491895"],"is_preprint":false},{"year":2025,"finding":"TGFβ1 activates JNK, which is recruited to the KLHL25 promoter by transcription factor NF-YA; JNK then phosphorylates histone H3 at Ser10 to activate KLHL25 transcription. Upon IL-6 signaling, ERK phosphorylates NF-YA, abolishing its DNA-binding ability and shutting off KLHL25 transcription, thereby preventing ACLY ubiquitination and degradation and supporting Th17 differentiation over iTreg.","method":"Chromatin immunoprecipitation (ChIP) showing JNK and NF-YA recruitment to KLHL25 promoter; phosphorylation assays; promoter reporter assays; Th17/iTreg differentiation assays with pathway inhibitors","journal":"Communications biology","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods (ChIP, phosphorylation assays, functional differentiation assays) establishing transcriptional regulation mechanism","pmids":["40119138"],"is_preprint":false}],"current_model":"KLHL25 functions as the substrate-recognition adaptor for the CUL3-RING E3 ubiquitin ligase complex, targeting hypophosphorylated 4E-BP1 and ACLY for ubiquitination and proteasomal degradation; KLHL25 transcription is activated by a TGFβ1-JNK-NF-YA-H3S10 phosphorylation axis and repressed by IL-6-ERK-mediated phosphorylation of NF-YA, thereby acting as a molecular switch governing fatty acid metabolism and the iTreg/Th17 cell fate decision."},"narrative":{"teleology":[{"year":2012,"claim":"Identifying KLHL25 as a CUL3 adaptor that targets hypophosphorylated 4E-BP1 for ubiquitin-dependent degradation established the first known function of the gene: homeostatic regulation of cap-dependent translation via eIF4E.","evidence":"Co-immunoprecipitation, RNAi knockdown, and in vitro ubiquitination assays in cultured cells","pmids":["22578813"],"confidence":"High","gaps":["Structural basis for selectivity toward hypophosphorylated versus phosphorylated 4E-BP1 is unknown","Physiological consequence of 4E-BP1 degradation in specific tissue or disease contexts not addressed","Whether additional CUL3-KLHL25 substrates exist was unresolved"]},{"year":2016,"claim":"Demonstrating that CUL3-KLHL25 also ubiquitinates ACLY for proteasomal degradation broadened the complex's substrate repertoire beyond translational control to lipid metabolism and showed tumor-suppressive relevance.","evidence":"Reciprocal co-immunoprecipitation, ubiquitination assays, and xenograft tumor growth assays","pmids":["27664236"],"confidence":"High","gaps":["The degron or binding interface on ACLY recognized by KLHL25 was not mapped","Whether KLHL25-dependent ACLY degradation operates in non-tumor cell types was untested","Relative contribution of 4E-BP1 versus ACLY degradation to KLHL25 tumor-suppressive activity was unclear"]},{"year":2021,"claim":"Linking TGFβ1-induced KLHL25-mediated ACLY degradation to a metabolic switch from fatty acid synthesis to oxidation explained how KLHL25 promotes iTreg differentiation by reducing malonyl-CoA and de-repressing CPT1.","evidence":"Ubiquitination assays, malonyl-CoA metabolomics, CPT1 activity measurement, and iTreg differentiation assays with KLHL25 knockdown/overexpression","pmids":["34491895"],"confidence":"High","gaps":["How TGFβ1 signaling upregulates KLHL25 expression was not resolved","Whether the 4E-BP1 substrate axis also contributes to T cell fate decisions was not tested","In vivo validation in conditional knockout mice was lacking"]},{"year":2025,"claim":"Elucidating the transcriptional regulation of KLHL25 — activated by TGFβ1-JNK-NF-YA-H3S10ph and repressed by IL-6-ERK-mediated NF-YA phosphorylation — revealed the upstream signaling switch that connects cytokine milieu to KLHL25 expression and iTreg/Th17 cell fate.","evidence":"ChIP for JNK and NF-YA at KLHL25 promoter, NF-YA phosphorylation assays, promoter reporter assays, Th17/iTreg differentiation with pathway inhibitors","pmids":["40119138"],"confidence":"High","gaps":["Whether NF-YA phosphorylation by ERK affects other target genes beyond KLHL25 was not dissected","In vivo confirmation of the JNK-NF-YA-H3S10ph axis at the KLHL25 locus in primary human T cells is lacking","Structural basis for KLHL25 substrate recognition (BTB-Kelch domain interactions) remains uncharacterized"]},{"year":null,"claim":"How the two known substrates (4E-BP1 and ACLY) are differentially selected by CUL3-KLHL25 in different cellular contexts, and whether additional substrates exist, remain open questions.","evidence":"","pmids":[],"confidence":"High","gaps":["No structural or degron-mapping study for either substrate–KLHL25 interaction","No conditional knockout mouse model to define in vivo requirements","Potential role of KLHL25 in tissues beyond immune cells and tumors is unexplored"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,1,2]}],"localization":[],"pathway":[{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[0,1,2]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[1,2,3]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[2,3]}],"complexes":["CUL3-KLHL25 E3 ubiquitin ligase"],"partners":["CUL3","EIF4EBP1","ACLY","NFYA"],"other_free_text":[]},"mechanistic_narrative":"KLHL25 is a substrate-recognition adaptor of the CUL3-RING E3 ubiquitin ligase complex that governs translational control and lipid metabolism by targeting specific substrates for proteasomal degradation. The CUL3-KLHL25 complex ubiquitinates hypophosphorylated 4E-BP1, providing homeostatic regulation of eIF4E-dependent translation [PMID:22578813], and also ubiquitinates ATP-citrate lyase (ACLY), thereby inhibiting de novo lipid synthesis [PMID:27664236]. During T helper cell differentiation, TGFβ1-induced KLHL25 expression promotes ACLY degradation, reducing malonyl-CoA levels and shifting fatty acid metabolism from synthesis to β-oxidation to support iTreg over Th17 fate; this transcriptional switch is driven by JNK-mediated histone H3S10 phosphorylation at the KLHL25 promoter via NF-YA, and is opposed by IL-6-ERK signaling, which phosphorylates NF-YA to silence KLHL25 [PMID:34491895, PMID:40119138]."},"prefetch_data":{"uniprot":{"accession":"Q9H0H3","full_name":"Kelch-like protein 25","aliases":["Ectoderm-neural cortex protein 2","ENC-2"],"length_aa":589,"mass_kda":65.9,"function":"Substrate-specific adapter of a BCR (BTB-CUL3-RBX1) E3 ubiquitin ligase complex involved in various processes, such as translation homeostasis and lipid synthesis (PubMed:22578813, PubMed:27664236, PubMed:34491895). The BCR(KLHL25) ubiquitin ligase complex acts by mediating ubiquitination of hypophosphorylated EIF4EBP1 (4E-BP1): ubiquitination and subsequent degradation of hypophosphorylated EIF4EBP1 (4E-BP1) probably serves as a homeostatic mechanism to maintain translation and prevent eIF4E inhibition when eIF4E levels are low (PubMed:22578813). The BCR(KLHL25) complex does not target EIF4EBP1 (4E-BP1) when it is hyperphosphorylated or associated with eIF4E (PubMed:22578813). The BCR(KLHL25) complex also acts as a regulator of lipid synthesis by mediating ubiquitination and degradation of ACLY, thereby inhibiting lipid synthesis (PubMed:27664236, PubMed:34491895). BCR(KLHL25)-mediated degradation of ACLY promotes fatty acid oxidation and is required for differentiation of inducible regulatory T (iTreg) cells (PubMed:34491895)","subcellular_location":"","url":"https://www.uniprot.org/uniprotkb/Q9H0H3/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/KLHL25","classification":"Not Classified","n_dependent_lines":4,"n_total_lines":1208,"dependency_fraction":0.0033112582781456954},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/KLHL25","total_profiled":1310},"omim":[{"mim_id":"619893","title":"KELCH-LIKE FAMILY, MEMBER 25; KLHL25","url":"https://www.omim.org/entry/619893"},{"mim_id":"603136","title":"CULLIN 3; CUL3","url":"https://www.omim.org/entry/603136"},{"mim_id":"108728","title":"ATP CITRATE LYASE; ACLY","url":"https://www.omim.org/entry/108728"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Nucleoplasm","reliability":"Approved"},{"location":"Cytosol","reliability":"Approved"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in many","driving_tissues":[],"url":"https://www.proteinatlas.org/search/KLHL25"},"hgnc":{"alias_symbol":["FLJ12587","ENC2","ENC-2"],"prev_symbol":[]},"alphafold":{"accession":"Q9H0H3","domains":[{"cath_id":"3.30.710.10","chopping":"29-111","consensus_level":"high","plddt":89.9164,"start":29,"end":111},{"cath_id":"1.25.40.420","chopping":"188-287","consensus_level":"high","plddt":83.9834,"start":188,"end":287},{"cath_id":"2.120.10.80","chopping":"294-583","consensus_level":"medium","plddt":89.5673,"start":294,"end":583}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9H0H3","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9H0H3-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9H0H3-F1-predicted_aligned_error_v6.png","plddt_mean":88.62},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=KLHL25","jax_strain_url":"https://www.jax.org/strain/search?query=KLHL25"},"sequence":{"accession":"Q9H0H3","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9H0H3.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9H0H3/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9H0H3"}},"corpus_meta":[{"pmid":"22578813","id":"PMC_22578813","title":"Translational homeostasis via the mRNA cap-binding protein, eIF4E.","date":"2012","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/22578813","citation_count":150,"is_preprint":false},{"pmid":"27664236","id":"PMC_27664236","title":"Cullin3-KLHL25 ubiquitin ligase targets ACLY for degradation to inhibit lipid synthesis and tumor progression.","date":"2016","source":"Genes & development","url":"https://pubmed.ncbi.nlm.nih.gov/27664236","citation_count":102,"is_preprint":false},{"pmid":"33751861","id":"PMC_33751861","title":"Exposure to violence, chronic stress, nasal DNA methylation, and atopic asthma in children.","date":"2021","source":"Pediatric pulmonology","url":"https://pubmed.ncbi.nlm.nih.gov/33751861","citation_count":33,"is_preprint":false},{"pmid":"30120715","id":"PMC_30120715","title":"Detection of Molecular Alterations in Taiwanese Patients with Medullary Thyroid Cancer Using Whole-Exome Sequencing.","date":"2018","source":"Endocrine pathology","url":"https://pubmed.ncbi.nlm.nih.gov/30120715","citation_count":32,"is_preprint":false},{"pmid":"34491895","id":"PMC_34491895","title":"ACLY ubiquitination by CUL3-KLHL25 induces the reprogramming of fatty acid metabolism to facilitate iTreg differentiation.","date":"2021","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/34491895","citation_count":31,"is_preprint":false},{"pmid":"6413277","id":"PMC_6413277","title":"The phenotype of engrailed mutations in the antenna of Drosophila.","date":"1983","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/6413277","citation_count":20,"is_preprint":false},{"pmid":"31122248","id":"PMC_31122248","title":"Exome-wide search and functional annotation of genes associated in patients with severe tick-borne encephalitis in a Russian population.","date":"2019","source":"BMC medical genomics","url":"https://pubmed.ncbi.nlm.nih.gov/31122248","citation_count":6,"is_preprint":false},{"pmid":"33173928","id":"PMC_33173928","title":"Exposure to violence, chronic stress, nasal DNA methylation, and atopic asthma in children.","date":"2020","source":"medRxiv : the preprint server for health sciences","url":"https://pubmed.ncbi.nlm.nih.gov/33173928","citation_count":5,"is_preprint":false},{"pmid":"39376559","id":"PMC_39376559","title":"Case report: Refractory focal motor seizure associated with cerebrospinal fluid neurochondrin antibody.","date":"2024","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/39376559","citation_count":1,"is_preprint":false},{"pmid":"40119138","id":"PMC_40119138","title":"KLHL25-ACLY module functions as a switch in the fate determination of the differentiation of iTreg/Th17.","date":"2025","source":"Communications biology","url":"https://pubmed.ncbi.nlm.nih.gov/40119138","citation_count":0,"is_preprint":false},{"pmid":"40672224","id":"PMC_40672224","title":"Identification of novel protective loci for executive function using the trail making test part B in the Long Life Family Study.","date":"2025","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/40672224","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":7173,"output_tokens":1161,"usd":0.019467},"stage2":{"model":"claude-opus-4-6","input_tokens":4366,"output_tokens":1718,"usd":0.09717},"total_usd":0.116637,"stage1_batch_id":"msgbatch_01Mgbec4vM6c9vXht7JVJzss","stage2_batch_id":"msgbatch_01JkuuH5RB2BdJNsgnhjbC6s","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2012,\n      \"finding\": \"KLHL25 forms a complex with CUL3 (KLHL25-CUL3) that acts as an E3 ubiquitin ligase targeting hypophosphorylated 4E-BP1 for ubiquitination and degradation, thereby providing homeostatic control of eIF4E activity.\",\n      \"method\": \"Biochemical identification of E3 ubiquitin ligase complex; knockdown experiments in cells showing 4E-BP1 degradation dependent on phosphorylation status\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods (RNAi knockdown, co-immunoprecipitation, ubiquitination assays) in a highly-cited foundational study\",\n      \"pmids\": [\"22578813\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"KLHL25 serves as the adaptor protein for the CUL3-RING ubiquitin ligase complex that binds ACLY (ATP-citrate lyase), mediating its ubiquitination and proteasomal degradation to inhibit lipid synthesis.\",\n      \"method\": \"Co-immunoprecipitation showing CUL3-KLHL25 interaction with ACLY; ubiquitination assays; xenograft tumor growth assays with loss-of-function\",\n      \"journal\": \"Genes & development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP, ubiquitination assay, and in vivo tumor model with functional readout in a well-cited study\",\n      \"pmids\": [\"27664236\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"TGFβ1 stimulation during iTreg differentiation induces CUL3-KLHL25-mediated ubiquitination and degradation of ACLY, which reduces malonyl-CoA levels, relieving inhibition of CPT1 and thereby shifting fatty acid metabolism from synthesis to oxidation to support iTreg differentiation.\",\n      \"method\": \"Biochemical ubiquitination assays, metabolic profiling (malonyl-CoA measurement), iTreg differentiation assays with KLHL25 knockdown/overexpression, CPT1 activity measurement\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods (ubiquitination assay, metabolomics, functional differentiation assay) in a single rigorous study\",\n      \"pmids\": [\"34491895\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"TGFβ1 activates JNK, which is recruited to the KLHL25 promoter by transcription factor NF-YA; JNK then phosphorylates histone H3 at Ser10 to activate KLHL25 transcription. Upon IL-6 signaling, ERK phosphorylates NF-YA, abolishing its DNA-binding ability and shutting off KLHL25 transcription, thereby preventing ACLY ubiquitination and degradation and supporting Th17 differentiation over iTreg.\",\n      \"method\": \"Chromatin immunoprecipitation (ChIP) showing JNK and NF-YA recruitment to KLHL25 promoter; phosphorylation assays; promoter reporter assays; Th17/iTreg differentiation assays with pathway inhibitors\",\n      \"journal\": \"Communications biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods (ChIP, phosphorylation assays, functional differentiation assays) establishing transcriptional regulation mechanism\",\n      \"pmids\": [\"40119138\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"KLHL25 functions as the substrate-recognition adaptor for the CUL3-RING E3 ubiquitin ligase complex, targeting hypophosphorylated 4E-BP1 and ACLY for ubiquitination and proteasomal degradation; KLHL25 transcription is activated by a TGFβ1-JNK-NF-YA-H3S10 phosphorylation axis and repressed by IL-6-ERK-mediated phosphorylation of NF-YA, thereby acting as a molecular switch governing fatty acid metabolism and the iTreg/Th17 cell fate decision.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"KLHL25 is a substrate-recognition adaptor of the CUL3-RING E3 ubiquitin ligase complex that governs translational control and lipid metabolism by targeting specific substrates for proteasomal degradation. The CUL3-KLHL25 complex ubiquitinates hypophosphorylated 4E-BP1, providing homeostatic regulation of eIF4E-dependent translation [PMID:22578813], and also ubiquitinates ATP-citrate lyase (ACLY), thereby inhibiting de novo lipid synthesis [PMID:27664236]. During T helper cell differentiation, TGFβ1-induced KLHL25 expression promotes ACLY degradation, reducing malonyl-CoA levels and shifting fatty acid metabolism from synthesis to β-oxidation to support iTreg over Th17 fate; this transcriptional switch is driven by JNK-mediated histone H3S10 phosphorylation at the KLHL25 promoter via NF-YA, and is opposed by IL-6-ERK signaling, which phosphorylates NF-YA to silence KLHL25 [PMID:34491895, PMID:40119138].\",\n  \"teleology\": [\n    {\n      \"year\": 2012,\n      \"claim\": \"Identifying KLHL25 as a CUL3 adaptor that targets hypophosphorylated 4E-BP1 for ubiquitin-dependent degradation established the first known function of the gene: homeostatic regulation of cap-dependent translation via eIF4E.\",\n      \"evidence\": \"Co-immunoprecipitation, RNAi knockdown, and in vitro ubiquitination assays in cultured cells\",\n      \"pmids\": [\"22578813\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Structural basis for selectivity toward hypophosphorylated versus phosphorylated 4E-BP1 is unknown\",\n        \"Physiological consequence of 4E-BP1 degradation in specific tissue or disease contexts not addressed\",\n        \"Whether additional CUL3-KLHL25 substrates exist was unresolved\"\n      ]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Demonstrating that CUL3-KLHL25 also ubiquitinates ACLY for proteasomal degradation broadened the complex's substrate repertoire beyond translational control to lipid metabolism and showed tumor-suppressive relevance.\",\n      \"evidence\": \"Reciprocal co-immunoprecipitation, ubiquitination assays, and xenograft tumor growth assays\",\n      \"pmids\": [\"27664236\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"The degron or binding interface on ACLY recognized by KLHL25 was not mapped\",\n        \"Whether KLHL25-dependent ACLY degradation operates in non-tumor cell types was untested\",\n        \"Relative contribution of 4E-BP1 versus ACLY degradation to KLHL25 tumor-suppressive activity was unclear\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Linking TGFβ1-induced KLHL25-mediated ACLY degradation to a metabolic switch from fatty acid synthesis to oxidation explained how KLHL25 promotes iTreg differentiation by reducing malonyl-CoA and de-repressing CPT1.\",\n      \"evidence\": \"Ubiquitination assays, malonyl-CoA metabolomics, CPT1 activity measurement, and iTreg differentiation assays with KLHL25 knockdown/overexpression\",\n      \"pmids\": [\"34491895\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"How TGFβ1 signaling upregulates KLHL25 expression was not resolved\",\n        \"Whether the 4E-BP1 substrate axis also contributes to T cell fate decisions was not tested\",\n        \"In vivo validation in conditional knockout mice was lacking\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Elucidating the transcriptional regulation of KLHL25 — activated by TGFβ1-JNK-NF-YA-H3S10ph and repressed by IL-6-ERK-mediated NF-YA phosphorylation — revealed the upstream signaling switch that connects cytokine milieu to KLHL25 expression and iTreg/Th17 cell fate.\",\n      \"evidence\": \"ChIP for JNK and NF-YA at KLHL25 promoter, NF-YA phosphorylation assays, promoter reporter assays, Th17/iTreg differentiation with pathway inhibitors\",\n      \"pmids\": [\"40119138\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether NF-YA phosphorylation by ERK affects other target genes beyond KLHL25 was not dissected\",\n        \"In vivo confirmation of the JNK-NF-YA-H3S10ph axis at the KLHL25 locus in primary human T cells is lacking\",\n        \"Structural basis for KLHL25 substrate recognition (BTB-Kelch domain interactions) remains uncharacterized\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the two known substrates (4E-BP1 and ACLY) are differentially selected by CUL3-KLHL25 in different cellular contexts, and whether additional substrates exist, remain open questions.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"No structural or degron-mapping study for either substrate–KLHL25 interaction\",\n        \"No conditional knockout mouse model to define in vivo requirements\",\n        \"Potential role of KLHL25 in tissues beyond immune cells and tumors is unexplored\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 1, 2]}\n    ],\n    \"localization\": [],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [0, 1, 2]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [1, 2, 3]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [2, 3]}\n    ],\n    \"complexes\": [\n      \"CUL3-KLHL25 E3 ubiquitin ligase\"\n    ],\n    \"partners\": [\n      \"CUL3\",\n      \"EIF4EBP1\",\n      \"ACLY\",\n      \"NFYA\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}