{"gene":"CLIC5","run_date":"2026-06-09T22:57:18","timeline":{"discoveries":[{"year":2000,"finding":"CLIC5 was isolated from placental microvilli as a component of a multimeric complex containing actin, ezrin, alpha-actinin, gelsolin, and IQGAP1; it associates with the detergent-insoluble cytoskeletal fraction of microvilli and localizes to the apical region of trophoblasts, distinguishing it from CLIC1 and CLIC4.","method":"Biochemical fractionation (detergent-insoluble fraction), co-isolation/pulldown, immunoblot, indirect immunofluorescence microscopy","journal":"Molecular biology of the cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-isolation with multiple cytoskeletal proteins from native tissue and fractionation, single lab, two orthogonal methods","pmids":["10793131"],"is_preprint":false},{"year":2006,"finding":"CLIC5 is expressed at high levels in the basal region of hair cell stereocilia (in approximate 1:1 molar ratio with radixin), and loss-of-function (jitterbug mutation causing premature stop codon) results in dysmorphic stereocilia, progressive hair cell degeneration, impaired hearing, and vestibular dysfunction; radixin immunostaining was reduced in hair bundles of mutant mice, suggesting CLIC5 associates with radixin to form or stabilize connections between the plasma membrane and the actin core.","method":"Spontaneous mouse mutant (jbg, 97 bp deletion/frameshift), immunohistochemistry, mass spectrometry, immunoblot, immunofluorescence","journal":"The Journal of neuroscience","confidence":"High","confidence_rationale":"Tier 2 / Strong — loss-of-function mouse model with defined cellular phenotype, mass spectrometry quantification, immunolocalization, multiple orthogonal methods","pmids":["17021174"],"is_preprint":false},{"year":2007,"finding":"Purified recombinant CLIC5 incorporated into planar lipid bilayers forms multiconductance ion channels that are almost equally permeable to Na+, K+, and Cl-; CLIC5 channel activity is strongly and reversibly inhibited by cytosolic F-actin in the absence of any other protein, an effect reversible by cytochalasin-mediated F-actin disruption.","method":"Planar lipid bilayer reconstitution with purified recombinant protein; cytochalasin treatment","journal":"The FEBS journal","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution in planar lipid bilayers with pharmacological validation (cytochalasin), single lab but multiple orthogonal conditions","pmids":["18028448"],"is_preprint":false},{"year":2010,"finding":"CLIC5 is enriched in podocyte foot processes, colocalizes and associates with the ezrin/radixin/moesin (ERM) complex and podocalyxin; CLIC5-knockout mice display decreased foot process length, widespread foot process abnormalities, proteinuria, and significantly decreased ERM complex and podocalyxin in podocytes.","method":"Proteomic purification of slit diaphragm fractions, co-immunoprecipitation, immunofluorescence, CLIC5 knockout mouse model with phenotypic readout (proteinuria, electron microscopy)","journal":"Kidney international","confidence":"High","confidence_rationale":"Tier 2 / Moderate — KO mouse with defined renal phenotype, co-immunoprecipitation, proteomic enrichment; multiple orthogonal methods, single lab","pmids":["20664558"],"is_preprint":false},{"year":2013,"finding":"CLIC5 forms a molecular complex with radixin (RDX), taperin (TPRN), and myosin VI (MYO6) at the base of stereocilia; loss of CLIC5 (jitterbug mice) causes progressive stereocilia fusion from postnatal day 10 and mislocalization of RDX, PTPRQ, and TPRN; CLIC5 and RDX also fail to localize normally in MYO6-mutant mice, placing CLIC5 in a complex that stabilizes membrane-actin linkages at the stereociliary base.","method":"Biochemical interaction assays (co-immunoprecipitation), confocal and scanning electron microscopy, mouse genetic models (jbg and MYO6 mutants), immunofluorescence","journal":"Cytoskeleton (Hoboken, N.J.)","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal genetic epistasis between CLIC5 and MYO6, biochemical interaction assays, multiple microscopy methods, defined cellular phenotype","pmids":["24285636"],"is_preprint":false},{"year":2015,"finding":"CLIC5 forms a complex with ezrin and podocalyxin in hepatocellular carcinoma cells; shRNA-mediated knockdown of CLIC5 in Huh7 cells decreases migration and invasion.","method":"shRNA knockdown, migration/invasion assays, Western blot, immunofluorescence","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — shRNA KD with functional phenotype (migration/invasion), single lab, but no detailed mechanistic dissection of the complex","pmids":["26135398"],"is_preprint":false},{"year":2016,"finding":"CLIC5 is a direct transcriptional target of ETV6 (identified by ChIP and transcriptome analysis); overexpression of CLIC5 increases resistance to hydrogen peroxide-induced apoptosis; CLIC5 ion channel activity modulates lysosomal-mediated cell death, possibly by affecting transferrin receptor function, with which CLIC5 colocalizes intracellularly.","method":"Chromatin immunoprecipitation (ChIP), whole transcriptome analysis, overexpression cell lines, apoptosis assays, colocalization by immunofluorescence","journal":"Haematologica","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP identifies direct target, functional overexpression assay, colocalization; single lab, mechanistic link to transferrin receptor is correlative","pmids":["27540136"],"is_preprint":false},{"year":2018,"finding":"KLPH/lctl is required for lens-specific expression of CLIC5; in KLPH knockout mice, CLIC5 transcripts are completely absent in the lens, and CLIC5 (detected by immunofluorescence) localizes to cilia/centrosomes of lens epithelial cells; jitterbug (Clic5-null) mice also exhibit defective lens sutures, implicating CLIC5 in lens fiber cell extension and organization.","method":"RNA-seq (knockout vs. wildtype), immunofluorescence localization, genetic mouse models (KLPH KO and jitterbug)","journal":"Experimental eye research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — RNA-seq and immunofluorescence in two genetic mouse models; CLIC5 localization to cilia/centrosomes established by direct imaging, single lab","pmids":["29425878"],"is_preprint":false},{"year":2021,"finding":"GRXCR2 interacts with CLIC5 at the base of stereocilia; deletion of the 60 amino acid N-terminal region of GRXCR2 required for CLIC5 interaction (by CRISPR/Cas9) causes moderate-to-severe hearing loss despite minimal effects on stereocilia morphogenesis, demonstrating that the GRXCR2–CLIC5 interaction is functionally important for hearing.","method":"Co-immunoprecipitation/interaction assay, CRISPR/Cas9 in-frame deletion, auditory function testing, immunolocalization","journal":"Frontiers in cell and developmental biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — CRISPR epistasis with defined auditory phenotype plus interaction assay; single lab, two orthogonal methods","pmids":["34026762"],"is_preprint":false},{"year":2023,"finding":"In zebrafish, CLIC5b isoform localizes to cilia and its knockdown causes defective ciliogenesis with ciliopathy-associated phenotypes (body curvature, otolith defects, altered left-right asymmetry, hydrocephalus, pronephric cysts) and dysregulation of cilia-dependent Wnt signaling; CLIC5a knockdown causes glomerular filtration barrier leakiness; mechanistically, Clic5 deficiency reduces phosphorylation of ERM (Ezrin/Radixin/Moesin) proteins in pronephric tubules.","method":"Morpholino/knockdown in zebrafish, whole-mount in situ hybridization, immunostaining, phenotypic analysis","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo knockdown with multiple defined phenotypes, ERM phosphorylation as mechanistic readout; single lab, zebrafish model","pmids":["37848494"],"is_preprint":false},{"year":2024,"finding":"CLIC5 promotes myogenic differentiation and skeletal muscle regeneration by interacting with biglycan (BGN) to activate the canonical Wnt/β-catenin signaling pathway; CLIC5 deletion reduces satellite cell number and differentiation, impairs Pax7 expression, and reduces BGN-mediated Wnt/β-catenin signaling, while CLIC5 overexpression enhances muscle regeneration.","method":"Muscle-specific CLIC5 knockout mice, overexpression in vivo and in vitro, co-immunoprecipitation (CLIC5-BGN interaction), Wnt/β-catenin pathway assays, satellite cell isolation","journal":"Science advances","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KO and overexpression mouse models with defined phenotype, co-immunoprecipitation of CLIC5-BGN, pathway assay; single lab, multiple orthogonal methods","pmids":["39999205"],"is_preprint":false},{"year":2010,"finding":"Constitutive overexpression of CLIC5 in C2C12 myoblasts reduces cell proliferation (G2/M to G0/G1 shift) and increases myotube size and number with upregulation of myosin heavy chain, myogenin, and desmin; CLIC5 protein expression increases during differentiation induced by medium switch.","method":"Overexpression in C2C12 cells, cell cycle analysis, immunoblot for myogenic markers","journal":"Cell biology international","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single overexpression experiment with phenotypic readout, no mechanistic pathway placement, single lab","pmids":["20055760"],"is_preprint":false},{"year":2022,"finding":"Using thermal shift assay screening of ~500 natural compounds, curcumin was identified as a specific interactant of purified recombinant CLIC5, as evidenced by dose-dependent effects on CLIC5 thermal stability.","method":"Differential scanning fluorimetry (thermal shift assay) with purified recombinant CLIC5","journal":"The Israel Medical Association journal : IMAJ","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single in vitro biophysical assay, no functional validation of the interaction, single lab","pmids":["37584386"],"is_preprint":false},{"year":2026,"finding":"CLIC5 is localized to the inner mitochondrial membrane in cardiomyoblasts and shows spatial proximity to TSPO (translocator protein) as measured by FRET (24% efficiency, comparable to positive control at 28%), suggesting TSPO may modulate CLIC5 (candidate for the centum-pS mitochondrial chloride channel) activity.","method":"FRET (acceptor photobleaching) in immunolabeled cardiomyoblasts","journal":"Biochemical and biophysical research communications","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single FRET measurement, no functional validation of the interaction, single lab","pmids":["41764818"],"is_preprint":false}],"current_model":"CLIC5 is a metamorphic chloride intracellular channel protein that can insert into membranes to form ion channels whose activity is directly inhibited by F-actin; it constitutively associates with ERM-family proteins (ezrin, radixin), actin-binding proteins (alpha-actinin, gelsolin), IQGAP1, taperin, myosin VI, and biglycan at actin-rich membrane specializations (microvilli, stereocilia base, podocyte foot processes, cilia), where it stabilizes plasma membrane–actin cytoskeleton linkages; loss of CLIC5 causes progressive stereocilia fusion, hair cell degeneration, deafness, vestibular dysfunction, podocyte foot process abnormalities, and impaired skeletal muscle regeneration, the latter mediated through biglycan-dependent activation of canonical Wnt/β-catenin signaling."},"narrative":{"mechanistic_narrative":"CLIC5 is a chloride intracellular channel protein that links the plasma membrane to the cortical actin cytoskeleton at actin-rich membrane specializations, where it stabilizes membrane–actin junctions critical for the integrity of microvilli, stereocilia, and podocyte foot processes [PMID:10793131, PMID:17021174, PMID:20664558]. It was first isolated from placental microvilli as part of a detergent-insoluble cytoskeletal complex containing actin, ezrin, alpha-actinin, gelsolin, and IQGAP1 [PMID:10793131], and it constitutively associates with ERM-family proteins (radixin, ezrin) and podocalyxin in both stereocilia and podocytes [PMID:17021174, PMID:20664558]. Purified recombinant CLIC5 inserts into planar lipid bilayers to form multiconductance channels permeable to Na+, K+, and Cl-, whose activity is strongly and reversibly inhibited by F-actin in the absence of any other protein, directly coupling channel function to cytoskeletal state [PMID:18028448]. At the stereociliary base, CLIC5 forms a complex with radixin, taperin, and myosin VI, and its loss causes progressive stereocilia fusion, mislocalization of partner proteins, hair cell degeneration, deafness, and vestibular dysfunction [PMID:17021174, PMID:24285636]; the functionally important GRXCR2–CLIC5 interaction at this site is also required for normal hearing [PMID:34026762]. In podocytes, CLIC5 loss reduces the ERM complex and podocalyxin and produces foot process abnormalities and proteinuria [PMID:20664558]. CLIC5 also localizes to cilia/centrosomes and supports ciliogenesis and cilia-dependent signaling, with deficiency reducing ERM phosphorylation [PMID:29425878, PMID:37848494]. Beyond its cytoskeletal scaffolding role, CLIC5 promotes myogenic differentiation and skeletal muscle regeneration by interacting with biglycan to activate canonical Wnt/β-catenin signaling [PMID:39999205].","teleology":[{"year":2000,"claim":"Established CLIC5 as a cytoskeleton-associated component of actin-rich membrane specializations rather than a free cytosolic protein, distinguishing it from other CLIC family members.","evidence":"Biochemical fractionation and co-isolation from placental microvilli with immunofluorescence localization","pmids":["10793131"],"confidence":"Medium","gaps":["Did not establish direct binary interactions within the complex","No channel function demonstrated","Functional consequence of complex assembly unknown"]},{"year":2006,"claim":"Linked CLIC5 loss-of-function to a defined cellular and organismal phenotype, showing it is required to maintain stereocilia and hearing and associates with radixin.","evidence":"Spontaneous jitterbug mouse mutant with immunohistochemistry, mass spectrometry, and immunolocalization","pmids":["17021174"],"confidence":"High","gaps":["Direct CLIC5–radixin binding not biochemically isolated","Whether channel activity is required for the phenotype untested"]},{"year":2007,"claim":"Demonstrated that CLIC5 is an intrinsic membrane-inserting ion channel whose activity is directly gated by F-actin, providing a molecular basis for coupling membrane conductance to cytoskeletal state.","evidence":"Planar lipid bilayer reconstitution with purified recombinant protein and cytochalasin treatment","pmids":["18028448"],"confidence":"High","gaps":["Physiological ion selectivity in vivo unresolved","Structural transition for membrane insertion not defined","Link between channel activity and tissue phenotypes not established"]},{"year":2010,"claim":"Extended CLIC5's membrane–cytoskeleton scaffolding role to podocytes, showing it stabilizes the ERM complex and podocalyxin to maintain foot process architecture and the filtration barrier.","evidence":"Slit diaphragm proteomics, co-immunoprecipitation, and CLIC5-knockout mouse with proteinuria and EM phenotype","pmids":["20664558"],"confidence":"High","gaps":["Mechanism by which CLIC5 stabilizes ERM/podocalyxin unresolved","Role of channel activity vs. scaffolding not separated"]},{"year":2013,"claim":"Placed CLIC5 within a defined radixin–taperin–myosin VI complex and showed genetic epistasis with MYO6, establishing it as part of a membrane-actin linkage machine at the stereociliary base.","evidence":"Co-immunoprecipitation, confocal/scanning EM, and jbg and MYO6 mutant mouse models","pmids":["24285636"],"confidence":"High","gaps":["Order of complex assembly and direct binding interfaces not mapped","Whether channel function contributes to linkage stability untested"]},{"year":2015,"claim":"Suggested CLIC5–ezrin–podocalyxin complexes promote cell migration and invasion in a cancer context, extending the scaffolding role to motility.","evidence":"shRNA knockdown with migration/invasion assays in Huh7 hepatocellular carcinoma cells","pmids":["26135398"],"confidence":"Medium","gaps":["No mechanistic dissection of the complex","Off-target effects of shRNA not excluded","Generalizability beyond one cell line unknown"]},{"year":2016,"claim":"Identified CLIC5 as a direct ETV6 transcriptional target and linked its channel activity to lysosomal cell death and oxidative stress resistance.","evidence":"ChIP, transcriptome analysis, overexpression apoptosis assays, and colocalization with transferrin receptor","pmids":["27540136"],"confidence":"Medium","gaps":["Transferrin receptor link is correlative","Mechanism connecting channel activity to lysosomal death undefined"]},{"year":2018,"claim":"Showed CLIC5 expression in the lens is controlled by KLPH/lctl and that CLIC5 localizes to cilia/centrosomes, implicating it in lens fiber organization.","evidence":"RNA-seq and immunofluorescence in KLPH-knockout and jitterbug mouse models","pmids":["29425878"],"confidence":"Medium","gaps":["Function of ciliary/centrosomal CLIC5 in lens not mechanistically defined","Whether lens phenotype reflects channel or scaffolding role unknown"]},{"year":2021,"claim":"Demonstrated that a specific GRXCR2–CLIC5 interaction interface is functionally required for hearing independent of gross stereocilia morphogenesis.","evidence":"Co-immunoprecipitation plus CRISPR/Cas9 in-frame deletion of the interaction domain with auditory testing","pmids":["34026762"],"confidence":"Medium","gaps":["Molecular consequence of losing the interaction at the stereociliary base unresolved","Whether GRXCR2 recruits or stabilizes CLIC5 not determined"]},{"year":2023,"claim":"Connected CLIC5 to ciliogenesis and ERM phosphorylation in vivo, linking ciliary CLIC5 to cilia-dependent Wnt signaling and the glomerular filtration barrier.","evidence":"Morpholino knockdown of CLIC5a/CLIC5b isoforms in zebrafish with phenotypic and immunostaining analysis","pmids":["37848494"],"confidence":"Medium","gaps":["Morpholino off-target effects not fully excluded","Direct mechanism connecting CLIC5 to ERM phosphorylation undefined","Isoform-specific functions only partially separated"]},{"year":2024,"claim":"Defined a signaling role for CLIC5 in muscle, showing it interacts with biglycan to activate canonical Wnt/β-catenin signaling driving satellite cell differentiation and regeneration.","evidence":"Muscle-specific knockout and overexpression mice, CLIC5–BGN co-immunoprecipitation, and Wnt/β-catenin pathway assays","pmids":["39999205"],"confidence":"Medium","gaps":["How CLIC5–BGN binding activates Wnt signaling mechanistically unresolved","Whether channel activity contributes untested"]},{"year":2010,"claim":"Provided early evidence that CLIC5 expression promotes myogenic differentiation while suppressing proliferation.","evidence":"Constitutive overexpression in C2C12 myoblasts with cell cycle analysis and myogenic marker immunoblots","pmids":["20055760"],"confidence":"Low","gaps":["Single overexpression experiment with no pathway placement","Endogenous loss-of-function not tested"]},{"year":2022,"claim":"Identified curcumin as a direct biophysical interactant of CLIC5, a candidate chemical probe.","evidence":"Thermal shift assay (differential scanning fluorimetry) screening with purified recombinant CLIC5","pmids":["37584386"],"confidence":"Low","gaps":["No functional validation of the interaction","Binding site and biological relevance unknown"]},{"year":2026,"claim":"Suggested a mitochondrial localization and TSPO proximity for CLIC5, raising the possibility it underlies a mitochondrial chloride conductance.","evidence":"FRET acceptor photobleaching in immunolabeled cardiomyoblasts","pmids":["41764818"],"confidence":"Low","gaps":["Single FRET measurement without functional validation","Identity as the centum-pS mitochondrial channel not established","Functional consequence of TSPO proximity unknown"]},{"year":null,"claim":"It remains unresolved how CLIC5's reconstituted ion channel activity relates mechanistically to its scaffolding functions in vivo, and whether channel conductance is required for any of the documented tissue phenotypes.","evidence":"No timeline study separates channel activity from membrane–cytoskeleton scaffolding in a physiological setting","pmids":[],"confidence":"Low","gaps":["No structure of the membrane-inserted channel","Channel-dead mutant phenotypes not reported","Relationship between cytosolic scaffold and inserted channel states undefined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0005215","term_label":"transporter activity","supporting_discovery_ids":[2]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[0,2]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[3,4]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0,1,3]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[0,2]},{"term_id":"GO:0005929","term_label":"cilium","supporting_discovery_ids":[7,9]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[7]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[10]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[10]}],"complexes":["CLIC5–radixin–taperin–myosin VI stereociliary complex","ERM–podocalyxin podocyte complex","actin/ezrin/alpha-actinin/gelsolin/IQGAP1 microvillar complex"],"partners":["RDX","EZR","PODXL","TPRN","MYO6","GRXCR2","BGN","IQGAP1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9NZA1","full_name":"Chloride intracellular channel protein 5","aliases":["Glutaredoxin-like oxidoreductase CLIC5"],"length_aa":410,"mass_kda":46.5,"function":"In the soluble state, catalyzes glutaredoxin-like thiol disulfide exchange reactions with reduced glutathione as electron donor (By similarity). Can insert into membranes and form non-selective ion channels almost equally permeable to Na(+), K(+) and Cl(-) (PubMed:15184393, PubMed:18028448). Required for normal hearing (PubMed:24781754). It is necessary for the formation of stereocilia in the inner ear and normal development of the organ of Corti (By similarity). May play a role in the regulation of transepithelial ion absorption and secretion. Is required for the development and/or maintenance of the proper glomerular endothelial cell and podocyte architecture (PubMed:15184393, PubMed:18028448, PubMed:20335315). Plays a role in formation of the lens suture in the eye, which is important for normal optical properties of the lens (By similarity)","subcellular_location":"Golgi apparatus; Cytoplasm, cytoskeleton, microtubule organizing center, centrosome","url":"https://www.uniprot.org/uniprotkb/Q9NZA1/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/CLIC5","classification":"Not Classified","n_dependent_lines":2,"n_total_lines":1208,"dependency_fraction":0.0016556291390728477},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/CLIC5","total_profiled":1310},"omim":[{"mim_id":"616042","title":"DEAFNESS, AUTOSOMAL RECESSIVE 103; DFNB103","url":"https://www.omim.org/entry/616042"},{"mim_id":"607293","title":"CHLORIDE INTRACELLULAR CHANNEL 5; CLIC5","url":"https://www.omim.org/entry/607293"},{"mim_id":"606536","title":"CHLORIDE INTRACELLULAR CHANNEL 4; CLIC4","url":"https://www.omim.org/entry/606536"},{"mim_id":"602872","title":"CHLORIDE INTRACELLULAR CHANNEL 1; CLIC1","url":"https://www.omim.org/entry/602872"},{"mim_id":"600074","title":"CD24 ANTIGEN; CD24","url":"https://www.omim.org/entry/600074"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Plasma membrane","reliability":"Supported"},{"location":"Nuclear speckles","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"heart muscle","ntpm":96.2},{"tissue":"skeletal muscle","ntpm":271.2},{"tissue":"tongue","ntpm":91.9}],"url":"https://www.proteinatlas.org/search/CLIC5"},"hgnc":{"alias_symbol":["DFNB102"],"prev_symbol":[]},"alphafold":{"accession":"Q9NZA1","domains":[{"cath_id":"3.40.30.10","chopping":"140-148_173-264","consensus_level":"medium","plddt":87.3576,"start":140,"end":264},{"cath_id":"1.20.1050.10","chopping":"266-317_333-410","consensus_level":"medium","plddt":93.7958,"start":266,"end":410}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NZA1","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NZA1-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NZA1-F1-predicted_aligned_error_v6.png","plddt_mean":68.0},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CLIC5","jax_strain_url":"https://www.jax.org/strain/search?query=CLIC5"},"sequence":{"accession":"Q9NZA1","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9NZA1.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9NZA1/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NZA1"}},"corpus_meta":[{"pmid":"10793131","id":"PMC_10793131","title":"Identification of a novel member of the chloride intracellular channel gene family (CLIC5) that associates with the actin cytoskeleton of placental microvilli.","date":"2000","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/10793131","citation_count":141,"is_preprint":false},{"pmid":"17021174","id":"PMC_17021174","title":"The chloride intracellular channel protein CLIC5 is expressed at high levels in hair cell stereocilia and is essential for normal inner ear function.","date":"2006","source":"The Journal of neuroscience : the official journal of the Society for Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/17021174","citation_count":109,"is_preprint":false},{"pmid":"18028448","id":"PMC_18028448","title":"Functional reconstitution of mammalian 'chloride intracellular channels' CLIC1, CLIC4 and CLIC5 reveals differential regulation by cytoskeletal actin.","date":"2007","source":"The FEBS journal","url":"https://pubmed.ncbi.nlm.nih.gov/18028448","citation_count":93,"is_preprint":false},{"pmid":"24285636","id":"PMC_24285636","title":"CLIC5 stabilizes membrane-actin filament linkages at the base of hair cell stereocilia in a molecular complex with radixin, taperin, and myosin VI.","date":"2013","source":"Cytoskeleton (Hoboken, N.J.)","url":"https://pubmed.ncbi.nlm.nih.gov/24285636","citation_count":64,"is_preprint":false},{"pmid":"20664558","id":"PMC_20664558","title":"Proteomic analysis of the slit diaphragm complex: CLIC5 is a protein critical for podocyte morphology and function.","date":"2010","source":"Kidney international","url":"https://pubmed.ncbi.nlm.nih.gov/20664558","citation_count":56,"is_preprint":false},{"pmid":"24781754","id":"PMC_24781754","title":"Progressive hearing loss and vestibular dysfunction caused by a homozygous nonsense mutation in CLIC5.","date":"2014","source":"European journal of human genetics : EJHG","url":"https://pubmed.ncbi.nlm.nih.gov/24781754","citation_count":50,"is_preprint":false},{"pmid":"26135398","id":"PMC_26135398","title":"Co-Expression of Ezrin-CLIC5-Podocalyxin Is Associated with Migration and Invasiveness in Hepatocellular Carcinoma.","date":"2015","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/26135398","citation_count":43,"is_preprint":false},{"pmid":"27540136","id":"PMC_27540136","title":"CLIC5: a novel ETV6 target gene in childhood acute lymphoblastic leukemia.","date":"2016","source":"Haematologica","url":"https://pubmed.ncbi.nlm.nih.gov/27540136","citation_count":23,"is_preprint":false},{"pmid":"29425878","id":"PMC_29425878","title":"The klotho-related protein KLPH (lctl) has preferred expression in lens and is essential for expression of clic5 and normal lens suture formation.","date":"2018","source":"Experimental eye research","url":"https://pubmed.ncbi.nlm.nih.gov/29425878","citation_count":21,"is_preprint":false},{"pmid":"39999205","id":"PMC_39999205","title":"CLIC5 promotes myoblast differentiation and skeletal muscle regeneration via the BGN-mediated canonical Wnt/β-catenin signaling pathway.","date":"2024","source":"Science advances","url":"https://pubmed.ncbi.nlm.nih.gov/39999205","citation_count":19,"is_preprint":false},{"pmid":"20055760","id":"PMC_20055760","title":"The CLIC5 (chloride intracellular channel 5) involved in C2C12 myoblasts proliferation and differentiation.","date":"2010","source":"Cell biology international","url":"https://pubmed.ncbi.nlm.nih.gov/20055760","citation_count":18,"is_preprint":false},{"pmid":"33114113","id":"PMC_33114113","title":"Bi-Allelic Novel Variants in CLIC5 Identified in a Cameroonian Multiplex Family with Non-Syndromic Hearing Impairment.","date":"2020","source":"Genes","url":"https://pubmed.ncbi.nlm.nih.gov/33114113","citation_count":17,"is_preprint":false},{"pmid":"34026762","id":"PMC_34026762","title":"N-Terminus of GRXCR2 Interacts With CLIC5 and Is Essential for Auditory Perception.","date":"2021","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/34026762","citation_count":11,"is_preprint":false},{"pmid":"37848494","id":"PMC_37848494","title":"A novel role for the chloride intracellular channel protein Clic5 in ciliary function.","date":"2023","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/37848494","citation_count":9,"is_preprint":false},{"pmid":"12200217","id":"PMC_12200217","title":"Identification and mutational analysis of candidate genes for juvenile myoclonic epilepsy on 6p11-p12: LRRC1, GCLC, KIAA0057 and CLIC5.","date":"2002","source":"Epilepsy research","url":"https://pubmed.ncbi.nlm.nih.gov/12200217","citation_count":9,"is_preprint":false},{"pmid":"38583737","id":"PMC_38583737","title":"Lnc-Clic5 as a sponge for miR-212-5p to inhibit cow barn PM2.5-induced apoptosis in rat alveolar macrophages.","date":"2024","source":"Toxicology","url":"https://pubmed.ncbi.nlm.nih.gov/38583737","citation_count":8,"is_preprint":false},{"pmid":"40962327","id":"PMC_40962327","title":"AQP1/4, CLIC5 Dysregulation and lipid metabolism alterations in lung cancer.","date":"2025","source":"BMB reports","url":"https://pubmed.ncbi.nlm.nih.gov/40962327","citation_count":3,"is_preprint":false},{"pmid":"40859056","id":"PMC_40859056","title":"AAV gene therapy rescues hearing and balance in a model of CLIC5 deafness.","date":"2025","source":"EMBO molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/40859056","citation_count":2,"is_preprint":false},{"pmid":"36635257","id":"PMC_36635257","title":"EPS8 variant causes deafness, autosomal recessive 102 (DFNB102) and literature review.","date":"2023","source":"Human genome variation","url":"https://pubmed.ncbi.nlm.nih.gov/36635257","citation_count":1,"is_preprint":false},{"pmid":"37584386","id":"PMC_37584386","title":"High-throughput analysis of CLIC5 interactants using a thermal-stability assay.","date":"2022","source":"The Israel Medical Association journal : IMAJ","url":"https://pubmed.ncbi.nlm.nih.gov/37584386","citation_count":0,"is_preprint":false},{"pmid":"41764818","id":"PMC_41764818","title":"Translocator protein in touch with mitochondrial chloride intracellular channel CLIC5.","date":"2026","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/41764818","citation_count":0,"is_preprint":false},{"pmid":"40957967","id":"PMC_40957967","title":"The c.644 G > A p.(Trp215*) founder variant in the CLIC5 gene causes progressive autosomal recessive deafness 103 (DFNB103) in Eastern Siberia.","date":"2025","source":"Journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/40957967","citation_count":0,"is_preprint":false},{"pmid":"41923498","id":"PMC_41923498","title":"Human CLIC5 as a Recurrent Hotspot of HPV 16 Integration in Cervical Cancer.","date":"2026","source":"Journal of medical virology","url":"https://pubmed.ncbi.nlm.nih.gov/41923498","citation_count":0,"is_preprint":false},{"pmid":"41913087","id":"PMC_41913087","title":"Exclusion of CLIC5 as a Candidate Gene and Identification of NEFM as a Possible Novel Gene Correlated With Autosomal Recessive Pure Cerebellar Ataxia in a Highly Consanguineous Family.","date":"2026","source":"Molecular genetics & genomic medicine","url":"https://pubmed.ncbi.nlm.nih.gov/41913087","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":13709,"output_tokens":3693,"usd":0.048261,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11409,"output_tokens":4461,"usd":0.084285,"stage2_stop_reason":"end_turn"},"total_usd":0.132546,"stage1_batch_id":"msgbatch_01ACAxJQabDsht39JXeWG22d","stage2_batch_id":"msgbatch_01QEU22A9U5sbPTgsMzTmvSf","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2000,\n      \"finding\": \"CLIC5 was isolated from placental microvilli as a component of a multimeric complex containing actin, ezrin, alpha-actinin, gelsolin, and IQGAP1; it associates with the detergent-insoluble cytoskeletal fraction of microvilli and localizes to the apical region of trophoblasts, distinguishing it from CLIC1 and CLIC4.\",\n      \"method\": \"Biochemical fractionation (detergent-insoluble fraction), co-isolation/pulldown, immunoblot, indirect immunofluorescence microscopy\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-isolation with multiple cytoskeletal proteins from native tissue and fractionation, single lab, two orthogonal methods\",\n      \"pmids\": [\"10793131\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"CLIC5 is expressed at high levels in the basal region of hair cell stereocilia (in approximate 1:1 molar ratio with radixin), and loss-of-function (jitterbug mutation causing premature stop codon) results in dysmorphic stereocilia, progressive hair cell degeneration, impaired hearing, and vestibular dysfunction; radixin immunostaining was reduced in hair bundles of mutant mice, suggesting CLIC5 associates with radixin to form or stabilize connections between the plasma membrane and the actin core.\",\n      \"method\": \"Spontaneous mouse mutant (jbg, 97 bp deletion/frameshift), immunohistochemistry, mass spectrometry, immunoblot, immunofluorescence\",\n      \"journal\": \"The Journal of neuroscience\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — loss-of-function mouse model with defined cellular phenotype, mass spectrometry quantification, immunolocalization, multiple orthogonal methods\",\n      \"pmids\": [\"17021174\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Purified recombinant CLIC5 incorporated into planar lipid bilayers forms multiconductance ion channels that are almost equally permeable to Na+, K+, and Cl-; CLIC5 channel activity is strongly and reversibly inhibited by cytosolic F-actin in the absence of any other protein, an effect reversible by cytochalasin-mediated F-actin disruption.\",\n      \"method\": \"Planar lipid bilayer reconstitution with purified recombinant protein; cytochalasin treatment\",\n      \"journal\": \"The FEBS journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution in planar lipid bilayers with pharmacological validation (cytochalasin), single lab but multiple orthogonal conditions\",\n      \"pmids\": [\"18028448\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"CLIC5 is enriched in podocyte foot processes, colocalizes and associates with the ezrin/radixin/moesin (ERM) complex and podocalyxin; CLIC5-knockout mice display decreased foot process length, widespread foot process abnormalities, proteinuria, and significantly decreased ERM complex and podocalyxin in podocytes.\",\n      \"method\": \"Proteomic purification of slit diaphragm fractions, co-immunoprecipitation, immunofluorescence, CLIC5 knockout mouse model with phenotypic readout (proteinuria, electron microscopy)\",\n      \"journal\": \"Kidney international\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO mouse with defined renal phenotype, co-immunoprecipitation, proteomic enrichment; multiple orthogonal methods, single lab\",\n      \"pmids\": [\"20664558\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"CLIC5 forms a molecular complex with radixin (RDX), taperin (TPRN), and myosin VI (MYO6) at the base of stereocilia; loss of CLIC5 (jitterbug mice) causes progressive stereocilia fusion from postnatal day 10 and mislocalization of RDX, PTPRQ, and TPRN; CLIC5 and RDX also fail to localize normally in MYO6-mutant mice, placing CLIC5 in a complex that stabilizes membrane-actin linkages at the stereociliary base.\",\n      \"method\": \"Biochemical interaction assays (co-immunoprecipitation), confocal and scanning electron microscopy, mouse genetic models (jbg and MYO6 mutants), immunofluorescence\",\n      \"journal\": \"Cytoskeleton (Hoboken, N.J.)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal genetic epistasis between CLIC5 and MYO6, biochemical interaction assays, multiple microscopy methods, defined cellular phenotype\",\n      \"pmids\": [\"24285636\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"CLIC5 forms a complex with ezrin and podocalyxin in hepatocellular carcinoma cells; shRNA-mediated knockdown of CLIC5 in Huh7 cells decreases migration and invasion.\",\n      \"method\": \"shRNA knockdown, migration/invasion assays, Western blot, immunofluorescence\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — shRNA KD with functional phenotype (migration/invasion), single lab, but no detailed mechanistic dissection of the complex\",\n      \"pmids\": [\"26135398\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"CLIC5 is a direct transcriptional target of ETV6 (identified by ChIP and transcriptome analysis); overexpression of CLIC5 increases resistance to hydrogen peroxide-induced apoptosis; CLIC5 ion channel activity modulates lysosomal-mediated cell death, possibly by affecting transferrin receptor function, with which CLIC5 colocalizes intracellularly.\",\n      \"method\": \"Chromatin immunoprecipitation (ChIP), whole transcriptome analysis, overexpression cell lines, apoptosis assays, colocalization by immunofluorescence\",\n      \"journal\": \"Haematologica\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP identifies direct target, functional overexpression assay, colocalization; single lab, mechanistic link to transferrin receptor is correlative\",\n      \"pmids\": [\"27540136\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"KLPH/lctl is required for lens-specific expression of CLIC5; in KLPH knockout mice, CLIC5 transcripts are completely absent in the lens, and CLIC5 (detected by immunofluorescence) localizes to cilia/centrosomes of lens epithelial cells; jitterbug (Clic5-null) mice also exhibit defective lens sutures, implicating CLIC5 in lens fiber cell extension and organization.\",\n      \"method\": \"RNA-seq (knockout vs. wildtype), immunofluorescence localization, genetic mouse models (KLPH KO and jitterbug)\",\n      \"journal\": \"Experimental eye research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — RNA-seq and immunofluorescence in two genetic mouse models; CLIC5 localization to cilia/centrosomes established by direct imaging, single lab\",\n      \"pmids\": [\"29425878\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"GRXCR2 interacts with CLIC5 at the base of stereocilia; deletion of the 60 amino acid N-terminal region of GRXCR2 required for CLIC5 interaction (by CRISPR/Cas9) causes moderate-to-severe hearing loss despite minimal effects on stereocilia morphogenesis, demonstrating that the GRXCR2–CLIC5 interaction is functionally important for hearing.\",\n      \"method\": \"Co-immunoprecipitation/interaction assay, CRISPR/Cas9 in-frame deletion, auditory function testing, immunolocalization\",\n      \"journal\": \"Frontiers in cell and developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — CRISPR epistasis with defined auditory phenotype plus interaction assay; single lab, two orthogonal methods\",\n      \"pmids\": [\"34026762\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"In zebrafish, CLIC5b isoform localizes to cilia and its knockdown causes defective ciliogenesis with ciliopathy-associated phenotypes (body curvature, otolith defects, altered left-right asymmetry, hydrocephalus, pronephric cysts) and dysregulation of cilia-dependent Wnt signaling; CLIC5a knockdown causes glomerular filtration barrier leakiness; mechanistically, Clic5 deficiency reduces phosphorylation of ERM (Ezrin/Radixin/Moesin) proteins in pronephric tubules.\",\n      \"method\": \"Morpholino/knockdown in zebrafish, whole-mount in situ hybridization, immunostaining, phenotypic analysis\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo knockdown with multiple defined phenotypes, ERM phosphorylation as mechanistic readout; single lab, zebrafish model\",\n      \"pmids\": [\"37848494\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CLIC5 promotes myogenic differentiation and skeletal muscle regeneration by interacting with biglycan (BGN) to activate the canonical Wnt/β-catenin signaling pathway; CLIC5 deletion reduces satellite cell number and differentiation, impairs Pax7 expression, and reduces BGN-mediated Wnt/β-catenin signaling, while CLIC5 overexpression enhances muscle regeneration.\",\n      \"method\": \"Muscle-specific CLIC5 knockout mice, overexpression in vivo and in vitro, co-immunoprecipitation (CLIC5-BGN interaction), Wnt/β-catenin pathway assays, satellite cell isolation\",\n      \"journal\": \"Science advances\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO and overexpression mouse models with defined phenotype, co-immunoprecipitation of CLIC5-BGN, pathway assay; single lab, multiple orthogonal methods\",\n      \"pmids\": [\"39999205\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Constitutive overexpression of CLIC5 in C2C12 myoblasts reduces cell proliferation (G2/M to G0/G1 shift) and increases myotube size and number with upregulation of myosin heavy chain, myogenin, and desmin; CLIC5 protein expression increases during differentiation induced by medium switch.\",\n      \"method\": \"Overexpression in C2C12 cells, cell cycle analysis, immunoblot for myogenic markers\",\n      \"journal\": \"Cell biology international\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single overexpression experiment with phenotypic readout, no mechanistic pathway placement, single lab\",\n      \"pmids\": [\"20055760\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Using thermal shift assay screening of ~500 natural compounds, curcumin was identified as a specific interactant of purified recombinant CLIC5, as evidenced by dose-dependent effects on CLIC5 thermal stability.\",\n      \"method\": \"Differential scanning fluorimetry (thermal shift assay) with purified recombinant CLIC5\",\n      \"journal\": \"The Israel Medical Association journal : IMAJ\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single in vitro biophysical assay, no functional validation of the interaction, single lab\",\n      \"pmids\": [\"37584386\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"CLIC5 is localized to the inner mitochondrial membrane in cardiomyoblasts and shows spatial proximity to TSPO (translocator protein) as measured by FRET (24% efficiency, comparable to positive control at 28%), suggesting TSPO may modulate CLIC5 (candidate for the centum-pS mitochondrial chloride channel) activity.\",\n      \"method\": \"FRET (acceptor photobleaching) in immunolabeled cardiomyoblasts\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single FRET measurement, no functional validation of the interaction, single lab\",\n      \"pmids\": [\"41764818\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"CLIC5 is a metamorphic chloride intracellular channel protein that can insert into membranes to form ion channels whose activity is directly inhibited by F-actin; it constitutively associates with ERM-family proteins (ezrin, radixin), actin-binding proteins (alpha-actinin, gelsolin), IQGAP1, taperin, myosin VI, and biglycan at actin-rich membrane specializations (microvilli, stereocilia base, podocyte foot processes, cilia), where it stabilizes plasma membrane–actin cytoskeleton linkages; loss of CLIC5 causes progressive stereocilia fusion, hair cell degeneration, deafness, vestibular dysfunction, podocyte foot process abnormalities, and impaired skeletal muscle regeneration, the latter mediated through biglycan-dependent activation of canonical Wnt/β-catenin signaling.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CLIC5 is a chloride intracellular channel protein that links the plasma membrane to the cortical actin cytoskeleton at actin-rich membrane specializations, where it stabilizes membrane–actin junctions critical for the integrity of microvilli, stereocilia, and podocyte foot processes [#0, #1, #3]. It was first isolated from placental microvilli as part of a detergent-insoluble cytoskeletal complex containing actin, ezrin, alpha-actinin, gelsolin, and IQGAP1 [#0], and it constitutively associates with ERM-family proteins (radixin, ezrin) and podocalyxin in both stereocilia and podocytes [#1, #3]. Purified recombinant CLIC5 inserts into planar lipid bilayers to form multiconductance channels permeable to Na+, K+, and Cl-, whose activity is strongly and reversibly inhibited by F-actin in the absence of any other protein, directly coupling channel function to cytoskeletal state [#2]. At the stereociliary base, CLIC5 forms a complex with radixin, taperin, and myosin VI, and its loss causes progressive stereocilia fusion, mislocalization of partner proteins, hair cell degeneration, deafness, and vestibular dysfunction [#1, #4]; the functionally important GRXCR2–CLIC5 interaction at this site is also required for normal hearing [#8]. In podocytes, CLIC5 loss reduces the ERM complex and podocalyxin and produces foot process abnormalities and proteinuria [#3]. CLIC5 also localizes to cilia/centrosomes and supports ciliogenesis and cilia-dependent signaling, with deficiency reducing ERM phosphorylation [#7, #9]. Beyond its cytoskeletal scaffolding role, CLIC5 promotes myogenic differentiation and skeletal muscle regeneration by interacting with biglycan to activate canonical Wnt/β-catenin signaling [#10].\",\n  \"teleology\": [\n    {\n      \"year\": 2000,\n      \"claim\": \"Established CLIC5 as a cytoskeleton-associated component of actin-rich membrane specializations rather than a free cytosolic protein, distinguishing it from other CLIC family members.\",\n      \"evidence\": \"Biochemical fractionation and co-isolation from placental microvilli with immunofluorescence localization\",\n      \"pmids\": [\"10793131\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Did not establish direct binary interactions within the complex\", \"No channel function demonstrated\", \"Functional consequence of complex assembly unknown\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Linked CLIC5 loss-of-function to a defined cellular and organismal phenotype, showing it is required to maintain stereocilia and hearing and associates with radixin.\",\n      \"evidence\": \"Spontaneous jitterbug mouse mutant with immunohistochemistry, mass spectrometry, and immunolocalization\",\n      \"pmids\": [\"17021174\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct CLIC5–radixin binding not biochemically isolated\", \"Whether channel activity is required for the phenotype untested\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Demonstrated that CLIC5 is an intrinsic membrane-inserting ion channel whose activity is directly gated by F-actin, providing a molecular basis for coupling membrane conductance to cytoskeletal state.\",\n      \"evidence\": \"Planar lipid bilayer reconstitution with purified recombinant protein and cytochalasin treatment\",\n      \"pmids\": [\"18028448\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological ion selectivity in vivo unresolved\", \"Structural transition for membrane insertion not defined\", \"Link between channel activity and tissue phenotypes not established\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Extended CLIC5's membrane–cytoskeleton scaffolding role to podocytes, showing it stabilizes the ERM complex and podocalyxin to maintain foot process architecture and the filtration barrier.\",\n      \"evidence\": \"Slit diaphragm proteomics, co-immunoprecipitation, and CLIC5-knockout mouse with proteinuria and EM phenotype\",\n      \"pmids\": [\"20664558\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which CLIC5 stabilizes ERM/podocalyxin unresolved\", \"Role of channel activity vs. scaffolding not separated\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Placed CLIC5 within a defined radixin–taperin–myosin VI complex and showed genetic epistasis with MYO6, establishing it as part of a membrane-actin linkage machine at the stereociliary base.\",\n      \"evidence\": \"Co-immunoprecipitation, confocal/scanning EM, and jbg and MYO6 mutant mouse models\",\n      \"pmids\": [\"24285636\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Order of complex assembly and direct binding interfaces not mapped\", \"Whether channel function contributes to linkage stability untested\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Suggested CLIC5–ezrin–podocalyxin complexes promote cell migration and invasion in a cancer context, extending the scaffolding role to motility.\",\n      \"evidence\": \"shRNA knockdown with migration/invasion assays in Huh7 hepatocellular carcinoma cells\",\n      \"pmids\": [\"26135398\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No mechanistic dissection of the complex\", \"Off-target effects of shRNA not excluded\", \"Generalizability beyond one cell line unknown\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Identified CLIC5 as a direct ETV6 transcriptional target and linked its channel activity to lysosomal cell death and oxidative stress resistance.\",\n      \"evidence\": \"ChIP, transcriptome analysis, overexpression apoptosis assays, and colocalization with transferrin receptor\",\n      \"pmids\": [\"27540136\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Transferrin receptor link is correlative\", \"Mechanism connecting channel activity to lysosomal death undefined\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Showed CLIC5 expression in the lens is controlled by KLPH/lctl and that CLIC5 localizes to cilia/centrosomes, implicating it in lens fiber organization.\",\n      \"evidence\": \"RNA-seq and immunofluorescence in KLPH-knockout and jitterbug mouse models\",\n      \"pmids\": [\"29425878\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Function of ciliary/centrosomal CLIC5 in lens not mechanistically defined\", \"Whether lens phenotype reflects channel or scaffolding role unknown\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Demonstrated that a specific GRXCR2–CLIC5 interaction interface is functionally required for hearing independent of gross stereocilia morphogenesis.\",\n      \"evidence\": \"Co-immunoprecipitation plus CRISPR/Cas9 in-frame deletion of the interaction domain with auditory testing\",\n      \"pmids\": [\"34026762\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular consequence of losing the interaction at the stereociliary base unresolved\", \"Whether GRXCR2 recruits or stabilizes CLIC5 not determined\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Connected CLIC5 to ciliogenesis and ERM phosphorylation in vivo, linking ciliary CLIC5 to cilia-dependent Wnt signaling and the glomerular filtration barrier.\",\n      \"evidence\": \"Morpholino knockdown of CLIC5a/CLIC5b isoforms in zebrafish with phenotypic and immunostaining analysis\",\n      \"pmids\": [\"37848494\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Morpholino off-target effects not fully excluded\", \"Direct mechanism connecting CLIC5 to ERM phosphorylation undefined\", \"Isoform-specific functions only partially separated\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Defined a signaling role for CLIC5 in muscle, showing it interacts with biglycan to activate canonical Wnt/β-catenin signaling driving satellite cell differentiation and regeneration.\",\n      \"evidence\": \"Muscle-specific knockout and overexpression mice, CLIC5–BGN co-immunoprecipitation, and Wnt/β-catenin pathway assays\",\n      \"pmids\": [\"39999205\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"How CLIC5–BGN binding activates Wnt signaling mechanistically unresolved\", \"Whether channel activity contributes untested\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Provided early evidence that CLIC5 expression promotes myogenic differentiation while suppressing proliferation.\",\n      \"evidence\": \"Constitutive overexpression in C2C12 myoblasts with cell cycle analysis and myogenic marker immunoblots\",\n      \"pmids\": [\"20055760\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Single overexpression experiment with no pathway placement\", \"Endogenous loss-of-function not tested\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Identified curcumin as a direct biophysical interactant of CLIC5, a candidate chemical probe.\",\n      \"evidence\": \"Thermal shift assay (differential scanning fluorimetry) screening with purified recombinant CLIC5\",\n      \"pmids\": [\"37584386\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No functional validation of the interaction\", \"Binding site and biological relevance unknown\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Suggested a mitochondrial localization and TSPO proximity for CLIC5, raising the possibility it underlies a mitochondrial chloride conductance.\",\n      \"evidence\": \"FRET acceptor photobleaching in immunolabeled cardiomyoblasts\",\n      \"pmids\": [\"41764818\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Single FRET measurement without functional validation\", \"Identity as the centum-pS mitochondrial channel not established\", \"Functional consequence of TSPO proximity unknown\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unresolved how CLIC5's reconstituted ion channel activity relates mechanistically to its scaffolding functions in vivo, and whether channel conductance is required for any of the documented tissue phenotypes.\",\n      \"evidence\": \"No timeline study separates channel activity from membrane–cytoskeleton scaffolding in a physiological setting\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No structure of the membrane-inserted channel\", \"Channel-dead mutant phenotypes not reported\", \"Relationship between cytosolic scaffold and inserted channel states undefined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0005215\", \"supporting_discovery_ids\": [2]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [0, 2]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [3, 4]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0, 1, 3]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [0, 2]},\n      {\"term_id\": \"GO:0005929\", \"supporting_discovery_ids\": [7, 9]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [7]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [10]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [10]}\n    ],\n    \"complexes\": [\n      \"CLIC5–radixin–taperin–myosin VI stereociliary complex\",\n      \"ERM–podocalyxin podocyte complex\",\n      \"actin/ezrin/alpha-actinin/gelsolin/IQGAP1 microvillar complex\"\n    ],\n    \"partners\": [\n      \"RDX\",\n      \"EZR\",\n      \"PODXL\",\n      \"TPRN\",\n      \"MYO6\",\n      \"GRXCR2\",\n      \"BGN\",\n      \"IQGAP1\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":7,"faith_pct":100.0}}