{"gene":"GET1","run_date":"2026-06-10T01:55:21","timeline":{"discoveries":[{"year":2011,"finding":"WRB (GET1 mammalian homolog) is an ER-resident membrane protein that acts as the receptor for TRC40/Asna1 (yeast Get3). The coiled-coil domain of WRB was identified as the binding site for TRC40/Asna1, and a soluble form of this domain interferes with TRC40/Asna1-mediated TA protein membrane insertion.","method":"Biochemical co-immunoprecipitation, cell imaging, dominant-negative soluble coiled-coil domain interference assay","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal biochemical binding assays with domain mapping and functional interference, replicated in subsequent studies","pmids":["21444755"],"is_preprint":false},{"year":2014,"finding":"The transmembrane domains of the Get1/2 (WRB/CAML) complex possess a bona fide insertase function distinct from their cytosolic domain functions: they capture the transmembrane domain of TA protein substrates and release them from Get3, defining a pre-integrated intermediate. Mutations in the Get1/2 transmembrane domains abolish TA protein insertion without disrupting Get3 binding by the cytosolic domains.","method":"Cell-based reporter assays and biochemical reconstitution with transmembrane domain mutants","journal":"Nature","confidence":"High","confidence_rationale":"Tier 1 / Strong — biochemical reconstitution combined with mutagenesis and cell reporters; single rigorous study with multiple orthogonal approaches","pmids":["25043001"],"is_preprint":false},{"year":2014,"finding":"WRB and CAML together are necessary and sufficient to constitute a functional ER membrane receptor for TRC40-mediated TA protein targeting. WRB and CAML expressed in yeast lacking Get1/Get2 rescue GET receptor mutant growth phenotypes and restore TA protein targeting. The membrane-spanning segments of CAML are essential for creating a functional receptor with WRB.","method":"Yeast complementation of GET1/GET2 deletion, in vivo TA protein targeting assays, binding parameter measurements for TRC40/WRB-CAML interaction","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic complementation in yeast plus quantitative binding measurements, replicated across labs","pmids":["24392163"],"is_preprint":false},{"year":2012,"finding":"The cytoplasmic domain of Get1 (Get1CD) stabilizes the open dimer conformation of Get3 ATPase by binding at two distinct interfaces simultaneously, promoting substrate release. Crystal structures of ADP-bound Get3 in complex with Get1CD were solved at 3.0 Å (open) and 4.5 Å (semi-open) resolution.","method":"X-ray crystallography of Get3–Get1CD complex; biochemical binding assays with interface mutants","journal":"Journal of molecular biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — crystal structure with biochemical validation, single lab with multiple orthogonal methods","pmids":["22684149"],"is_preprint":false},{"year":2017,"finding":"A single Get1/2 heterodimer is sufficient for TA protein insertion into the ER membrane. The conserved cytosolic regions of Get1 and Get2 bind asymmetrically to opposing subunits of the Get3 homodimer.","method":"Single-molecule and bulk fluorescence measurements in reconstituted lipid bilayers; quantitative in vitro insertion analysis","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 1 / Moderate — reconstituted system with single-molecule and ensemble methods defining stoichiometry and architecture","pmids":["28877464"],"is_preprint":false},{"year":2019,"finding":"WRB acts catalytically to assist the topogenesis of its partner CAML: in the presence of sufficient WRB, CAML is inserted into the ER membrane with three transmembrane segments in its C-terminal region. Without sufficient WRB, CAML fails to adopt the correct topology and instead generates aberrant topoforms that accumulate in ER-associated clusters and are degraded by the proteasome.","method":"Topology assays, proteasome inhibition, microscopy of ER-associated clusters in WRB-depleted cells","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — topology determination and degradation pathway experiments, single lab, two complementary methods","pmids":["31417168"],"is_preprint":false},{"year":2021,"finding":"Complex assembly between the cytosolic domains (CDs) of Get1 and Get2 strongly enhances the affinity of individual subunits for the Get3•TA targeting complex. Two molecular recognition features (MoRFs) in Get2CD induce Get3 opening, while Get1CD remodels Get3 conformation; both subunits are required for optimal TA release from Get3. Mutation of the MoRFs attenuates TA insertion in vivo.","method":"Fluorescence binding assays measuring Get1CD and Get2CD affinity for Get3•TA, single-molecule FRET, in vivo TA insertion assays with MoRF mutants","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — multiple orthogonal fluorescence methods plus in vivo mutant validation in a single rigorous study","pmids":["34614151"],"is_preprint":false},{"year":2022,"finding":"Get1/2 forms an aqueous channel (~2.5 nm diameter, corresponding to circumference of two Get1/2 complexes) in reconstituted bilayers. Get3 binding seals the Get1/2 channel, which dynamically opens and closes. Channel activity is required for releasing TA proteins from Get3 for membrane insertion; Get1/2 is proposed to also translocate C-terminal hydrophilic segments.","method":"Bulk fluorescence and microfluidics channel-forming assays in reconstituted bilayers; mutational analysis of channel activity correlated with TA protein insertion","journal":"Cell reports","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — reconstituted in vitro channel assay with mutagenesis, single lab, novel finding not yet independently replicated","pmids":["36640319"],"is_preprint":false},{"year":2016,"finding":"WRB mediates insertion of otoferlin (a TA protein essential for hair cell exocytosis) into the ER via the TRC40 pathway. Disruption of Wrb in zebrafish hair cells reduced otoferlin levels, impaired hearing, and caused defective synaptic vesicle replenishment. Transgenic Wrb rescue and otoferlin overexpression restored hearing in zebrafish. A WRB mutant (R73A) unable to bind Trc40 failed to rescue, establishing the TRC40-binding interface as functionally essential.","method":"Hair cell-specific Wrb knockout in zebrafish and mice; electrophysiology (patch-clamp), auditory recordings, immunohistochemistry, transgenic rescue including binding-defective mutant","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — loss-of-function in two organisms with defined cellular phenotype, mechanistic rescue with binding-defective mutant, multiple orthogonal readouts","pmids":["27458190"],"is_preprint":false},{"year":2016,"finding":"WRB (zebrafish wrb/pwi) is required for synaptic vesicle biogenesis at ribbon synapses in photoreceptors. Mutation of wrb reduced Rab3 and CSP/Dnajc5 in hair cells and photoreceptors, reduced ribbon number and vesicles surrounding ribbons, and abolished the optokinetic response. Morpholino knockdown of trc40 phenocopied wrb mutation, and overexpression of wrbR73A (cannot bind Trc40) failed to rescue, confirming that the Trc40-binding interface of WRB is required.","method":"ERG, optokinetic response, immunohistochemistry, electron microscopy in zebrafish wrb mutants; morpholino knockdown of trc40; rescue with wild-type vs. R73A mutant wrb","journal":"Investigative ophthalmology & visual science","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods including EM ultrastructure, electrophysiology, and mutant rescue experiments","pmids":["27273592"],"is_preprint":false},{"year":2016,"finding":"WRB knockout mice show that SNARE syntaxin 5 (Stx5) is extremely sensitive to TRC40 pathway disruption and is an autophagy target when the pathway is impaired. In contrast, other TA proteins showed differential sensitivity to WRB loss, demonstrating that in vitro TRC40-pathway client status does not predict in vivo dependence on the receptor.","method":"Tissue-specific WRB knockout mouse models; western blotting, TA protein fate assays; yeast screen of TA proteins combined with mammalian validation","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo knockout with substrate-specific mechanistic follow-up, single lab","pmids":["28000760"],"is_preprint":false},{"year":2018,"finding":"Get1/2 ER membrane insertase complex is required for efficient mitophagy during prolonged respiratory growth in yeast. This requirement is independent of Get3 (the cytosolic ATPase), as Get3-deficient cells show only slight mitophagy defects, suggesting that Get1/2-dependent TA protein(s) or the Get1/2 complex itself acts specifically in the mitophagy pathway downstream of Atg32.","method":"Yeast deletion mutants of get1/2 and get3; mitophagy assays during respiratory growth; Atg32 localization and expression analysis","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis between get1/2 and get3, specific mitophagy phenotype with Atg32-independent branch identified, single lab","pmids":["29673596"],"is_preprint":false},{"year":2003,"finding":"GET1/GRHL3 protein localizes to the nucleus, binds Grainyhead DNA-binding sites, homodimerizes via a short C-terminal domain, and contains a transactivation domain (aa 100–190) sufficient to confer transactivation to a heterologous GAL4 DBD. The DNA-binding domain maps to the region homologous to Drosophila Grainyhead DBD. GET1 also contains repression domains, consistent with dual activator/repressor function. GET1 was identified as an LMO-4 interacting partner by yeast two-hybrid.","method":"Yeast two-hybrid screen for LMO-4 partners; reporter assays for transactivation; dimerization assays; nuclear localization by cell imaging; Grainyhead site EMSA","journal":"Developmental dynamics","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — multiple functional assays (DNA binding, transactivation, dimerization, localization) in a single characterization study","pmids":["12666198"],"is_preprint":false},{"year":2008,"finding":"Get1/Grhl3 acts upstream of TGFα in the EGFR/ERK pathway during eyelid closure: Get1 knockout mice have reduced TGFα expression, reduced phospho-EGFR and phospho-ERK at the leading edge, and defective F-actin polymerization and filopodia formation. TGFα treatment in organ culture rescued cell shape changes and leading edge formation in Get1−/− eyelids.","method":"Get1 knockout mouse analysis; immunostaining for phospho-EGFR, phospho-ERK, F-actin; organ culture TGFα rescue experiment","journal":"Developmental biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis established by rescue experiment, multiple pathway readouts, single lab","pmids":["18485343"],"is_preprint":false},{"year":2009,"finding":"Get1/Grhl3 directly binds the uroplakin II promoter in urothelial cells and activates its transcription, driving urothelial differentiation and apical membrane specialization. This binding is regulated by histone modifications. Get1 knockout mice have defective bladder epithelial barrier due to failure of uroplakin-dependent apical membrane specialization.","method":"Get1 knockout mouse; genome-wide expression profiling; ChIP demonstrating Get1 binding to uroplakin II promoter; histone modification analysis","journal":"The EMBO journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP evidence for direct promoter binding plus in vivo knockout phenotype, single lab","pmids":["19494835"],"is_preprint":false},{"year":2012,"finding":"GRHL3/GET1 recruits the Trithorax complex to a subset of epidermal differentiation genes, activating their expression. GET1 cooperates with Trithorax group members both for genes that are Polycomb-repressed in progenitors and for Polycomb-independent differentiation genes.","method":"Genome-wide chromatin analysis, co-recruitment assays; Grhl3 knockout epidermis analysis","journal":"PLoS genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genome-wide binding data plus knockout with defined gene expression changes, single lab","pmids":["22829784"],"is_preprint":false},{"year":2012,"finding":"Grhl3/Get1 binds and represses the miR-21 promoter in the epidermis, establishing a regulatory loop. Loss of Grhl3 increases miR-21 levels, which in Ras-transformed keratinocytes leads to enhanced downregulation of MSH2 and other miR-21 targets, in part through downregulation of the RNA-binding protein DND1 during transformation.","method":"miRNA profiling in Grhl3−/− skin; ChIP showing Grhl3 binding to miR-21 promoter; reporter and overexpression assays; subcutaneous tumor formation assay","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP for direct promoter binding, miRNA profiling, and in vivo tumorigenesis assay, single lab","pmids":["22614019"],"is_preprint":false},{"year":1990,"finding":"The Wrb antigen on glycophorin A (GPA) requires the interaction of GPA with band 3 (SLC4A1) for its expression on erythrocytes. Wrb antibodies immunoprecipitate both band 3 and glycophorin A together, while monospecific antibodies to each protein precipitate only their cognate antigen, indicating a direct GPA–band 3 complex underlies Wrb antigen expression.","method":"Radioimmunoassay with monoclonal anti-Wrb antibodies; co-immunoprecipitation of erythrocyte membrane proteins","journal":"Blood","confidence":"Medium","confidence_rationale":"Tier 3 / Strong — co-immunoprecipitation replicated across multiple labs with consistent results","pmids":["2383660"],"is_preprint":false}],"current_model":"GET1/WRB is an ER-resident transmembrane protein that functions as part of the GET/TRC40 pathway for post-translational insertion of tail-anchored (TA) proteins: its coiled-coil cytosolic domain recruits the targeting ATPase Get3/TRC40 and, together with its partner CAML/Get2, stabilizes the open conformation of Get3 to release TA substrates, while its transmembrane segments form an aqueous insertase channel that captures TA transmembrane domains and facilitates their entry into the lipid bilayer; WRB also catalyzes correct topogenesis of CAML and is required for synaptic vesicle protein biogenesis in sensory hair cells and photoreceptors, mitophagy in yeast, and—as the mammalian transcription factor GRHL3/GET1—directly binds differentiation gene promoters, recruits Trithorax complexes, and represses miR-21 in epidermal cells."},"narrative":{"mechanistic_narrative":"The GET1 symbol in this corpus resolves to two distinct, internally coherent proteins: WRB, an ER-resident membrane insertase of the GET/TRC40 tail-anchored (TA) protein targeting pathway, and GRHL3/GET1, a nuclear Grainyhead-family transcription factor; both are well-evidenced and treated below. As WRB, the protein is an ER membrane receptor that, with its partner CAML/Get2, captures TRC40/Get3-delivered TA proteins and inserts them into the bilayer [PMID:21444755, PMID:24392163]. The cytosolic coiled-coil domain of Get1/WRB binds the TRC40/Get3 ATPase and, together with two molecular recognition features in the Get2/CAML cytosolic domain, stabilizes the open Get3 dimer conformation that drives substrate release [PMID:22684149, PMID:34614151]. Separately from this cytosolic recruitment function, the transmembrane segments of the Get1/2 (WRB/CAML) complex provide a bona fide insertase activity that captures the substrate TM domain and forms an aqueous channel which is gated by Get3 binding [PMID:25043001, PMID:36640319]; a single Get1/2 heterodimer suffices for insertion, with Get1 and Get2 binding asymmetrically to opposing Get3 subunits [PMID:28877464]. WRB further acts catalytically to enforce correct three-TM topogenesis of CAML, preventing accumulation of aberrant topoforms that are proteasomally degraded [PMID:31417168]. In vivo, WRB is required for biogenesis of synaptic-vesicle and TA proteins such as otoferlin and syntaxin 5 in sensory hair cells and photoreceptors, where loss disrupts ribbon synapse vesicle replenishment, hearing, and vision [PMID:27458190, PMID:27273592, PMID:28000760], and the Get1/2 complex supports mitophagy in yeast independently of Get3 [PMID:29673596]. As the transcription factor GRHL3/GET1, the protein localizes to the nucleus, binds Grainyhead DNA sites, dimerizes, and carries both activation and repression domains [PMID:12666198]; it directly binds and regulates differentiation gene promoters, recruiting Trithorax complexes [PMID:19494835, PMID:22829784], represses the miR-21 promoter in epidermis [PMID:22614019], and acts upstream of TGFα in EGFR/ERK signaling during eyelid closure [PMID:18485343]. An unrelated erythrocyte Wrb blood-group antigen arises from a glycophorin A–band 3 complex [PMID:2383660].","teleology":[{"year":1990,"claim":"Established the original molecular basis of the erythrocyte Wrb antigen, showing it depends on a glycophorin A–band 3 membrane complex rather than a single protein.","evidence":"Radioimmunoassay and co-immunoprecipitation of erythrocyte membrane proteins with monoclonal anti-Wrb antibodies","pmids":["2383660"],"confidence":"Medium","gaps":["Distinct from the ER insertase WRB and the transcription factor GET1; relationship to those proteins not addressed","No structural detail of the GPA–band 3 interface"]},{"year":2003,"claim":"Defined GET1/GRHL3 as a nuclear Grainyhead-family transcription factor with DNA-binding, dimerization, and dual activator/repressor domains, framing its role in epithelial gene regulation.","evidence":"Yeast two-hybrid for LMO-4 partners, EMSA, transactivation reporter assays, dimerization and localization assays","pmids":["12666198"],"confidence":"Medium","gaps":["Direct target genes not yet identified","No genome-wide binding map","Mechanism of repression-domain function unresolved"]},{"year":2008,"claim":"Placed Grhl3/Get1 upstream of EGFR/ERK signaling in morphogenesis by showing it controls TGFα expression required for leading-edge actin dynamics during eyelid closure.","evidence":"Get1 knockout mouse with phospho-EGFR/ERK and F-actin staining and TGFα organ-culture rescue","pmids":["18485343"],"confidence":"Medium","gaps":["Whether Get1 binds the TGFα promoter directly not established","Single lab"]},{"year":2009,"claim":"Demonstrated direct promoter binding by Grhl3/Get1, linking it to terminal epithelial differentiation through activation of uroplakin II and apical membrane specialization.","evidence":"ChIP, genome-wide expression profiling, and barrier-defect phenotype in Get1 knockout bladder","pmids":["19494835"],"confidence":"Medium","gaps":["Identity of cooperating chromatin factors not yet defined here","Histone-modification dependence described correlatively"]},{"year":2011,"claim":"Identified WRB as the ER membrane receptor for the TRC40/Asna1 TA-targeting ATPase, mapping the binding interface to its cytosolic coiled-coil domain.","evidence":"Co-immunoprecipitation, imaging, and dominant-negative soluble coiled-coil interference in cells","pmids":["21444755"],"confidence":"High","gaps":["Did not resolve how WRB drives substrate release or insertion","Role of a partner subunit not yet established"]},{"year":2012,"claim":"Provided the structural mechanism by which Get1 opens Get3, showing the Get1 cytosolic domain binds two interfaces of the Get3 dimer to promote substrate release.","evidence":"X-ray crystallography of Get3–Get1CD complexes plus interface-mutant binding assays","pmids":["22684149"],"confidence":"High","gaps":["Did not address insertase activity of the TM segments","Contribution of Get2 to opening not quantified here"]},{"year":2012,"claim":"Connected GRHL3/GET1 to chromatin machinery and miRNA control, showing it recruits Trithorax complexes to differentiation genes and represses miR-21 to restrain transformation.","evidence":"Genome-wide chromatin analysis, co-recruitment assays, ChIP, miRNA profiling, and tumor-formation assays in Grhl3 knockout skin","pmids":["22829784","22614019"],"confidence":"Medium","gaps":["Direct biochemical contact between GRHL3 and Trithorax subunits not mapped","Mechanism of miR-21 promoter repression not detailed"]},{"year":2014,"claim":"Separated the insertase function of the Get1/2 transmembrane domains from cytosolic Get3 recruitment, and showed WRB+CAML are necessary and sufficient to constitute the receptor.","evidence":"Cell reporter assays and biochemical reconstitution with TM-domain mutants; yeast complementation by WRB/CAML with binding measurements","pmids":["25043001","24392163"],"confidence":"High","gaps":["Physical nature of the insertion conduit not yet defined","Stoichiometry of the active complex unresolved"]},{"year":2016,"claim":"Established the physiological importance of WRB-dependent TA insertion in excitable sensory cells, with otoferlin and synaptic-vesicle proteins as key clients and the TRC40-binding interface as functionally essential.","evidence":"Hair-cell and photoreceptor Wrb knockout/mutant in zebrafish and mice with electrophysiology, EM, immunostaining, and R73A binding-defective rescue","pmids":["27458190","27273592"],"confidence":"High","gaps":["Full client repertoire in these cells not enumerated","Whether vesicle defects are entirely TA-insertion-dependent not fully resolved"]},{"year":2016,"claim":"Showed that in vitro client status does not predict in vivo receptor dependence, identifying syntaxin 5 as exquisitely WRB-sensitive and an autophagy target upon pathway failure.","evidence":"Tissue-specific WRB knockout mice with western blotting, TA-fate assays, and a yeast-to-mammalian client screen","pmids":["28000760"],"confidence":"Medium","gaps":["Determinants of differential substrate sensitivity unknown","Single lab"]},{"year":2018,"claim":"Revealed a Get3-independent role for the Get1/2 insertase in mitophagy, implicating a membrane-intrinsic function downstream of Atg32.","evidence":"Yeast get1/2 and get3 deletion mutants with mitophagy assays during respiratory growth","pmids":["29673596"],"confidence":"Medium","gaps":["The relevant TA substrate(s) for mitophagy not identified","Whether the role is conserved in mammals untested here"]},{"year":2021,"claim":"Quantified how Get1/2 complex assembly cooperatively enhances Get3•TA binding, defining Get2 MoRFs and Get1 remodeling as jointly required for substrate release.","evidence":"Fluorescence binding assays, single-molecule FRET, and in vivo insertion assays with MoRF mutants","pmids":["34614151"],"confidence":"High","gaps":["Precise ordering of conformational steps during release not fully resolved","Coupling to membrane insertion step not addressed"]},{"year":2022,"claim":"Demonstrated that Get1/2 forms a gated aqueous channel whose opening, sealed by Get3, is required for TA release and possibly translocation of C-terminal hydrophilic segments.","evidence":"Bulk fluorescence and microfluidics channel assays in reconstituted bilayers with channel-activity mutants","pmids":["36640319"],"confidence":"Medium","gaps":["Channel model not yet independently replicated","Direct evidence for C-terminal translocation not provided","No high-resolution channel structure"]},{"year":null,"claim":"It remains unresolved how the cytosolic substrate-release machinery is physically coupled to the transmembrane insertase/channel within a single Get1/2(WRB/CAML)–Get3 reaction cycle, and what governs substrate-specific receptor dependence in vivo.","evidence":"No single study in the timeline integrates the conformational opening, channel gating, and in vivo client-selectivity findings","pmids":[],"confidence":"Medium","gaps":["No integrated structure of the full membrane-embedded WRB/CAML–Get3–substrate intermediate","Rules predicting which TA proteins require WRB in vivo unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[1,5,7]},{"term_id":"GO:0005198","term_label":"structural molecule activity","supporting_discovery_ids":[1,4,7]},{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[12,14,15,16]},{"term_id":"GO:0003677","term_label":"DNA binding","supporting_discovery_ids":[12,14]},{"term_id":"GO:0005215","term_label":"transporter activity","supporting_discovery_ids":[7]}],"localization":[{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[0,1,2,5]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0,1,17]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[12,14]}],"pathway":[{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[0,1,2]},{"term_id":"R-HSA-9609507","term_label":"Protein localization","supporting_discovery_ids":[0,1,4]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[12,14,15,16]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[13,14]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[11]}],"complexes":["GET insertase complex (WRB/Get1–CAML/Get2)"],"partners":["TRC40","GET3","CAML","LMO4","SLC4A1","GYPA"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"O00258","full_name":"Guided entry of tail-anchored proteins factor 1","aliases":["Congenital heart disease 5 protein","Tail-anchored protein insertion receptor WRB","Tryptophan-rich basic protein"],"length_aa":174,"mass_kda":19.8,"function":"Required for the post-translational delivery of tail-anchored (TA) proteins to the endoplasmic reticulum (ER) (PubMed:21444755, PubMed:23041287, PubMed:24392163, PubMed:27226539). Together with CAMLG/GET2, acts as a membrane receptor for soluble GET3/TRC40, which recognizes and selectively binds the transmembrane domain of TA proteins in the cytosol (PubMed:21444755, PubMed:23041287, PubMed:24392163, PubMed:27226539). Required to ensure correct topology and ER insertion of CAMLG (PubMed:31417168, PubMed:32187542)","subcellular_location":"Endoplasmic reticulum membrane","url":"https://www.uniprot.org/uniprotkb/O00258/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/GET1","classification":"Not Classified","n_dependent_lines":601,"n_total_lines":1208,"dependency_fraction":0.49751655629139074},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"VAPA","stoichiometry":0.2},{"gene":"VCP","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/GET1","total_profiled":1310},"omim":[{"mim_id":"620781","title":"TRANSMEMBRANE PROTEIN 208; TMEM208","url":"https://www.omim.org/entry/620781"},{"mim_id":"612056","title":"GUIDED ENTRY OF TAIL-ANCHORED PROTEINS FACTOR 4; GET4","url":"https://www.omim.org/entry/612056"},{"mim_id":"602915","title":"GUIDED ENTRY OF TAIL-ANCHORED PROTEINS FACTOR 1; GET1","url":"https://www.omim.org/entry/602915"},{"mim_id":"601118","title":"CALCIUM-MODULATING LIGAND; CAMLG","url":"https://www.omim.org/entry/601118"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"choroid plexus","ntpm":135.4}],"url":"https://www.proteinatlas.org/search/GET1"},"hgnc":{"alias_symbol":["CHD5"],"prev_symbol":["WRB"]},"alphafold":{"accession":"O00258","domains":[{"cath_id":"-","chopping":"125-174","consensus_level":"medium","plddt":64.4726,"start":125,"end":174},{"cath_id":"1.10.287","chopping":"1-123","consensus_level":"medium","plddt":83.1118,"start":1,"end":123}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/O00258","model_url":"https://alphafold.ebi.ac.uk/files/AF-O00258-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-O00258-F1-predicted_aligned_error_v6.png","plddt_mean":77.69},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=GET1","jax_strain_url":"https://www.jax.org/strain/search?query=GET1"},"sequence":{"accession":"O00258","fasta_url":"https://rest.uniprot.org/uniprotkb/O00258.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/O00258/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/O00258"}},"corpus_meta":[{"pmid":"25043001","id":"PMC_25043001","title":"The Get1/2 transmembrane complex is an endoplasmic-reticulum membrane protein insertase.","date":"2014","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/25043001","citation_count":106,"is_preprint":false},{"pmid":"21444755","id":"PMC_21444755","title":"WRB is the receptor for TRC40/Asna1-mediated insertion of tail-anchored proteins into the ER membrane.","date":"2011","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/21444755","citation_count":98,"is_preprint":false},{"pmid":"22829784","id":"PMC_22829784","title":"GRHL3/GET1 and trithorax group members collaborate to activate the epidermal progenitor differentiation program.","date":"2012","source":"PLoS genetics","url":"https://pubmed.ncbi.nlm.nih.gov/22829784","citation_count":84,"is_preprint":false},{"pmid":"6342608","id":"PMC_6342608","title":"The Wrb antigen, a receptor for Plasmodium falciparum malaria, is located on a helical region of the major membrane sialoglycoprotein of human red blood cells.","date":"1983","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/6342608","citation_count":75,"is_preprint":false},{"pmid":"952768","id":"PMC_952768","title":"Anti-Wrb, and other autoantibodies responsible for positive direct antiglobulin tests in 150 individuals.","date":"1976","source":"British journal of haematology","url":"https://pubmed.ncbi.nlm.nih.gov/952768","citation_count":71,"is_preprint":false},{"pmid":"12666198","id":"PMC_12666198","title":"Identification and characterization of Grainyhead-like epithelial transactivator (GET-1), a novel mammalian Grainyhead-like factor.","date":"2003","source":"Developmental dynamics : an official publication of the American Association of Anatomists","url":"https://pubmed.ncbi.nlm.nih.gov/12666198","citation_count":69,"is_preprint":false},{"pmid":"19494835","id":"PMC_19494835","title":"The epidermal differentiation-associated Grainyhead gene Get1/Grhl3 also regulates urothelial differentiation.","date":"2009","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/19494835","citation_count":68,"is_preprint":false},{"pmid":"2383660","id":"PMC_2383660","title":"Relationship of the human erythrocyte Wrb antigen to an interaction between glycophorin A and band 3.","date":"1990","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/2383660","citation_count":65,"is_preprint":false},{"pmid":"22614019","id":"PMC_22614019","title":"The Grainyhead transcription factor Grhl3/Get1 suppresses miR-21 expression and tumorigenesis in skin: modulation of the miR-21 target MSH2 by RNA-binding protein DND1.","date":"2012","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/22614019","citation_count":62,"is_preprint":false},{"pmid":"24392163","id":"PMC_24392163","title":"WRB and CAML are necessary and sufficient to mediate tail-anchored protein targeting to the ER membrane.","date":"2014","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/24392163","citation_count":55,"is_preprint":false},{"pmid":"18485343","id":"PMC_18485343","title":"Grainyhead-like factor Get1/Grhl3 regulates formation of the epidermal leading edge during eyelid closure.","date":"2008","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/18485343","citation_count":53,"is_preprint":false},{"pmid":"3539140","id":"PMC_3539140","title":"High frequency antigens of human erythrocyte membrane sialoglycoproteins, III. 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The coiled-coil domain of WRB was identified as the binding site for TRC40/Asna1, and a soluble form of this domain interferes with TRC40/Asna1-mediated TA protein membrane insertion.\",\n      \"method\": \"Biochemical co-immunoprecipitation, cell imaging, dominant-negative soluble coiled-coil domain interference assay\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal biochemical binding assays with domain mapping and functional interference, replicated in subsequent studies\",\n      \"pmids\": [\"21444755\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"The transmembrane domains of the Get1/2 (WRB/CAML) complex possess a bona fide insertase function distinct from their cytosolic domain functions: they capture the transmembrane domain of TA protein substrates and release them from Get3, defining a pre-integrated intermediate. Mutations in the Get1/2 transmembrane domains abolish TA protein insertion without disrupting Get3 binding by the cytosolic domains.\",\n      \"method\": \"Cell-based reporter assays and biochemical reconstitution with transmembrane domain mutants\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — biochemical reconstitution combined with mutagenesis and cell reporters; single rigorous study with multiple orthogonal approaches\",\n      \"pmids\": [\"25043001\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"WRB and CAML together are necessary and sufficient to constitute a functional ER membrane receptor for TRC40-mediated TA protein targeting. WRB and CAML expressed in yeast lacking Get1/Get2 rescue GET receptor mutant growth phenotypes and restore TA protein targeting. The membrane-spanning segments of CAML are essential for creating a functional receptor with WRB.\",\n      \"method\": \"Yeast complementation of GET1/GET2 deletion, in vivo TA protein targeting assays, binding parameter measurements for TRC40/WRB-CAML interaction\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic complementation in yeast plus quantitative binding measurements, replicated across labs\",\n      \"pmids\": [\"24392163\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"The cytoplasmic domain of Get1 (Get1CD) stabilizes the open dimer conformation of Get3 ATPase by binding at two distinct interfaces simultaneously, promoting substrate release. Crystal structures of ADP-bound Get3 in complex with Get1CD were solved at 3.0 Å (open) and 4.5 Å (semi-open) resolution.\",\n      \"method\": \"X-ray crystallography of Get3–Get1CD complex; biochemical binding assays with interface mutants\",\n      \"journal\": \"Journal of molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structure with biochemical validation, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"22684149\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"A single Get1/2 heterodimer is sufficient for TA protein insertion into the ER membrane. The conserved cytosolic regions of Get1 and Get2 bind asymmetrically to opposing subunits of the Get3 homodimer.\",\n      \"method\": \"Single-molecule and bulk fluorescence measurements in reconstituted lipid bilayers; quantitative in vitro insertion analysis\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — reconstituted system with single-molecule and ensemble methods defining stoichiometry and architecture\",\n      \"pmids\": [\"28877464\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"WRB acts catalytically to assist the topogenesis of its partner CAML: in the presence of sufficient WRB, CAML is inserted into the ER membrane with three transmembrane segments in its C-terminal region. Without sufficient WRB, CAML fails to adopt the correct topology and instead generates aberrant topoforms that accumulate in ER-associated clusters and are degraded by the proteasome.\",\n      \"method\": \"Topology assays, proteasome inhibition, microscopy of ER-associated clusters in WRB-depleted cells\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — topology determination and degradation pathway experiments, single lab, two complementary methods\",\n      \"pmids\": [\"31417168\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Complex assembly between the cytosolic domains (CDs) of Get1 and Get2 strongly enhances the affinity of individual subunits for the Get3•TA targeting complex. Two molecular recognition features (MoRFs) in Get2CD induce Get3 opening, while Get1CD remodels Get3 conformation; both subunits are required for optimal TA release from Get3. Mutation of the MoRFs attenuates TA insertion in vivo.\",\n      \"method\": \"Fluorescence binding assays measuring Get1CD and Get2CD affinity for Get3•TA, single-molecule FRET, in vivo TA insertion assays with MoRF mutants\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — multiple orthogonal fluorescence methods plus in vivo mutant validation in a single rigorous study\",\n      \"pmids\": [\"34614151\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Get1/2 forms an aqueous channel (~2.5 nm diameter, corresponding to circumference of two Get1/2 complexes) in reconstituted bilayers. Get3 binding seals the Get1/2 channel, which dynamically opens and closes. Channel activity is required for releasing TA proteins from Get3 for membrane insertion; Get1/2 is proposed to also translocate C-terminal hydrophilic segments.\",\n      \"method\": \"Bulk fluorescence and microfluidics channel-forming assays in reconstituted bilayers; mutational analysis of channel activity correlated with TA protein insertion\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — reconstituted in vitro channel assay with mutagenesis, single lab, novel finding not yet independently replicated\",\n      \"pmids\": [\"36640319\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"WRB mediates insertion of otoferlin (a TA protein essential for hair cell exocytosis) into the ER via the TRC40 pathway. Disruption of Wrb in zebrafish hair cells reduced otoferlin levels, impaired hearing, and caused defective synaptic vesicle replenishment. Transgenic Wrb rescue and otoferlin overexpression restored hearing in zebrafish. A WRB mutant (R73A) unable to bind Trc40 failed to rescue, establishing the TRC40-binding interface as functionally essential.\",\n      \"method\": \"Hair cell-specific Wrb knockout in zebrafish and mice; electrophysiology (patch-clamp), auditory recordings, immunohistochemistry, transgenic rescue including binding-defective mutant\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — loss-of-function in two organisms with defined cellular phenotype, mechanistic rescue with binding-defective mutant, multiple orthogonal readouts\",\n      \"pmids\": [\"27458190\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"WRB (zebrafish wrb/pwi) is required for synaptic vesicle biogenesis at ribbon synapses in photoreceptors. Mutation of wrb reduced Rab3 and CSP/Dnajc5 in hair cells and photoreceptors, reduced ribbon number and vesicles surrounding ribbons, and abolished the optokinetic response. Morpholino knockdown of trc40 phenocopied wrb mutation, and overexpression of wrbR73A (cannot bind Trc40) failed to rescue, confirming that the Trc40-binding interface of WRB is required.\",\n      \"method\": \"ERG, optokinetic response, immunohistochemistry, electron microscopy in zebrafish wrb mutants; morpholino knockdown of trc40; rescue with wild-type vs. R73A mutant wrb\",\n      \"journal\": \"Investigative ophthalmology & visual science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods including EM ultrastructure, electrophysiology, and mutant rescue experiments\",\n      \"pmids\": [\"27273592\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"WRB knockout mice show that SNARE syntaxin 5 (Stx5) is extremely sensitive to TRC40 pathway disruption and is an autophagy target when the pathway is impaired. In contrast, other TA proteins showed differential sensitivity to WRB loss, demonstrating that in vitro TRC40-pathway client status does not predict in vivo dependence on the receptor.\",\n      \"method\": \"Tissue-specific WRB knockout mouse models; western blotting, TA protein fate assays; yeast screen of TA proteins combined with mammalian validation\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo knockout with substrate-specific mechanistic follow-up, single lab\",\n      \"pmids\": [\"28000760\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Get1/2 ER membrane insertase complex is required for efficient mitophagy during prolonged respiratory growth in yeast. This requirement is independent of Get3 (the cytosolic ATPase), as Get3-deficient cells show only slight mitophagy defects, suggesting that Get1/2-dependent TA protein(s) or the Get1/2 complex itself acts specifically in the mitophagy pathway downstream of Atg32.\",\n      \"method\": \"Yeast deletion mutants of get1/2 and get3; mitophagy assays during respiratory growth; Atg32 localization and expression analysis\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis between get1/2 and get3, specific mitophagy phenotype with Atg32-independent branch identified, single lab\",\n      \"pmids\": [\"29673596\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"GET1/GRHL3 protein localizes to the nucleus, binds Grainyhead DNA-binding sites, homodimerizes via a short C-terminal domain, and contains a transactivation domain (aa 100–190) sufficient to confer transactivation to a heterologous GAL4 DBD. The DNA-binding domain maps to the region homologous to Drosophila Grainyhead DBD. GET1 also contains repression domains, consistent with dual activator/repressor function. GET1 was identified as an LMO-4 interacting partner by yeast two-hybrid.\",\n      \"method\": \"Yeast two-hybrid screen for LMO-4 partners; reporter assays for transactivation; dimerization assays; nuclear localization by cell imaging; Grainyhead site EMSA\",\n      \"journal\": \"Developmental dynamics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — multiple functional assays (DNA binding, transactivation, dimerization, localization) in a single characterization study\",\n      \"pmids\": [\"12666198\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Get1/Grhl3 acts upstream of TGFα in the EGFR/ERK pathway during eyelid closure: Get1 knockout mice have reduced TGFα expression, reduced phospho-EGFR and phospho-ERK at the leading edge, and defective F-actin polymerization and filopodia formation. TGFα treatment in organ culture rescued cell shape changes and leading edge formation in Get1−/− eyelids.\",\n      \"method\": \"Get1 knockout mouse analysis; immunostaining for phospho-EGFR, phospho-ERK, F-actin; organ culture TGFα rescue experiment\",\n      \"journal\": \"Developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis established by rescue experiment, multiple pathway readouts, single lab\",\n      \"pmids\": [\"18485343\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Get1/Grhl3 directly binds the uroplakin II promoter in urothelial cells and activates its transcription, driving urothelial differentiation and apical membrane specialization. This binding is regulated by histone modifications. Get1 knockout mice have defective bladder epithelial barrier due to failure of uroplakin-dependent apical membrane specialization.\",\n      \"method\": \"Get1 knockout mouse; genome-wide expression profiling; ChIP demonstrating Get1 binding to uroplakin II promoter; histone modification analysis\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP evidence for direct promoter binding plus in vivo knockout phenotype, single lab\",\n      \"pmids\": [\"19494835\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"GRHL3/GET1 recruits the Trithorax complex to a subset of epidermal differentiation genes, activating their expression. GET1 cooperates with Trithorax group members both for genes that are Polycomb-repressed in progenitors and for Polycomb-independent differentiation genes.\",\n      \"method\": \"Genome-wide chromatin analysis, co-recruitment assays; Grhl3 knockout epidermis analysis\",\n      \"journal\": \"PLoS genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genome-wide binding data plus knockout with defined gene expression changes, single lab\",\n      \"pmids\": [\"22829784\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Grhl3/Get1 binds and represses the miR-21 promoter in the epidermis, establishing a regulatory loop. Loss of Grhl3 increases miR-21 levels, which in Ras-transformed keratinocytes leads to enhanced downregulation of MSH2 and other miR-21 targets, in part through downregulation of the RNA-binding protein DND1 during transformation.\",\n      \"method\": \"miRNA profiling in Grhl3−/− skin; ChIP showing Grhl3 binding to miR-21 promoter; reporter and overexpression assays; subcutaneous tumor formation assay\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP for direct promoter binding, miRNA profiling, and in vivo tumorigenesis assay, single lab\",\n      \"pmids\": [\"22614019\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1990,\n      \"finding\": \"The Wrb antigen on glycophorin A (GPA) requires the interaction of GPA with band 3 (SLC4A1) for its expression on erythrocytes. Wrb antibodies immunoprecipitate both band 3 and glycophorin A together, while monospecific antibodies to each protein precipitate only their cognate antigen, indicating a direct GPA–band 3 complex underlies Wrb antigen expression.\",\n      \"method\": \"Radioimmunoassay with monoclonal anti-Wrb antibodies; co-immunoprecipitation of erythrocyte membrane proteins\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Strong — co-immunoprecipitation replicated across multiple labs with consistent results\",\n      \"pmids\": [\"2383660\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"GET1/WRB is an ER-resident transmembrane protein that functions as part of the GET/TRC40 pathway for post-translational insertion of tail-anchored (TA) proteins: its coiled-coil cytosolic domain recruits the targeting ATPase Get3/TRC40 and, together with its partner CAML/Get2, stabilizes the open conformation of Get3 to release TA substrates, while its transmembrane segments form an aqueous insertase channel that captures TA transmembrane domains and facilitates their entry into the lipid bilayer; WRB also catalyzes correct topogenesis of CAML and is required for synaptic vesicle protein biogenesis in sensory hair cells and photoreceptors, mitophagy in yeast, and—as the mammalian transcription factor GRHL3/GET1—directly binds differentiation gene promoters, recruits Trithorax complexes, and represses miR-21 in epidermal cells.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"The GET1 symbol in this corpus resolves to two distinct, internally coherent proteins: WRB, an ER-resident membrane insertase of the GET/TRC40 tail-anchored (TA) protein targeting pathway, and GRHL3/GET1, a nuclear Grainyhead-family transcription factor; both are well-evidenced and treated below. As WRB, the protein is an ER membrane receptor that, with its partner CAML/Get2, captures TRC40/Get3-delivered TA proteins and inserts them into the bilayer [#0, #2]. The cytosolic coiled-coil domain of Get1/WRB binds the TRC40/Get3 ATPase and, together with two molecular recognition features in the Get2/CAML cytosolic domain, stabilizes the open Get3 dimer conformation that drives substrate release [#3, #6]. Separately from this cytosolic recruitment function, the transmembrane segments of the Get1/2 (WRB/CAML) complex provide a bona fide insertase activity that captures the substrate TM domain and forms an aqueous channel which is gated by Get3 binding [#1, #7]; a single Get1/2 heterodimer suffices for insertion, with Get1 and Get2 binding asymmetrically to opposing Get3 subunits [#4]. WRB further acts catalytically to enforce correct three-TM topogenesis of CAML, preventing accumulation of aberrant topoforms that are proteasomally degraded [#5]. In vivo, WRB is required for biogenesis of synaptic-vesicle and TA proteins such as otoferlin and syntaxin 5 in sensory hair cells and photoreceptors, where loss disrupts ribbon synapse vesicle replenishment, hearing, and vision [#8, #9, #10], and the Get1/2 complex supports mitophagy in yeast independently of Get3 [#11]. As the transcription factor GRHL3/GET1, the protein localizes to the nucleus, binds Grainyhead DNA sites, dimerizes, and carries both activation and repression domains [#12]; it directly binds and regulates differentiation gene promoters, recruiting Trithorax complexes [#14, #15], represses the miR-21 promoter in epidermis [#16], and acts upstream of TGF\\u03b1 in EGFR/ERK signaling during eyelid closure [#13]. An unrelated erythrocyte Wrb blood-group antigen arises from a glycophorin A\\u2013band 3 complex [#17].\",\n  \"teleology\": [\n    {\n      \"year\": 1990,\n      \"claim\": \"Established the original molecular basis of the erythrocyte Wrb antigen, showing it depends on a glycophorin A\\u2013band 3 membrane complex rather than a single protein.\",\n      \"evidence\": \"Radioimmunoassay and co-immunoprecipitation of erythrocyte membrane proteins with monoclonal anti-Wrb antibodies\",\n      \"pmids\": [\"2383660\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Distinct from the ER insertase WRB and the transcription factor GET1; relationship to those proteins not addressed\", \"No structural detail of the GPA\\u2013band 3 interface\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Defined GET1/GRHL3 as a nuclear Grainyhead-family transcription factor with DNA-binding, dimerization, and dual activator/repressor domains, framing its role in epithelial gene regulation.\",\n      \"evidence\": \"Yeast two-hybrid for LMO-4 partners, EMSA, transactivation reporter assays, dimerization and localization assays\",\n      \"pmids\": [\"12666198\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct target genes not yet identified\", \"No genome-wide binding map\", \"Mechanism of repression-domain function unresolved\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Placed Grhl3/Get1 upstream of EGFR/ERK signaling in morphogenesis by showing it controls TGF\\u03b1 expression required for leading-edge actin dynamics during eyelid closure.\",\n      \"evidence\": \"Get1 knockout mouse with phospho-EGFR/ERK and F-actin staining and TGF\\u03b1 organ-culture rescue\",\n      \"pmids\": [\"18485343\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether Get1 binds the TGF\\u03b1 promoter directly not established\", \"Single lab\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Demonstrated direct promoter binding by Grhl3/Get1, linking it to terminal epithelial differentiation through activation of uroplakin II and apical membrane specialization.\",\n      \"evidence\": \"ChIP, genome-wide expression profiling, and barrier-defect phenotype in Get1 knockout bladder\",\n      \"pmids\": [\"19494835\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Identity of cooperating chromatin factors not yet defined here\", \"Histone-modification dependence described correlatively\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Identified WRB as the ER membrane receptor for the TRC40/Asna1 TA-targeting ATPase, mapping the binding interface to its cytosolic coiled-coil domain.\",\n      \"evidence\": \"Co-immunoprecipitation, imaging, and dominant-negative soluble coiled-coil interference in cells\",\n      \"pmids\": [\"21444755\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not resolve how WRB drives substrate release or insertion\", \"Role of a partner subunit not yet established\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Provided the structural mechanism by which Get1 opens Get3, showing the Get1 cytosolic domain binds two interfaces of the Get3 dimer to promote substrate release.\",\n      \"evidence\": \"X-ray crystallography of Get3\\u2013Get1CD complexes plus interface-mutant binding assays\",\n      \"pmids\": [\"22684149\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not address insertase activity of the TM segments\", \"Contribution of Get2 to opening not quantified here\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Connected GRHL3/GET1 to chromatin machinery and miRNA control, showing it recruits Trithorax complexes to differentiation genes and represses miR-21 to restrain transformation.\",\n      \"evidence\": \"Genome-wide chromatin analysis, co-recruitment assays, ChIP, miRNA profiling, and tumor-formation assays in Grhl3 knockout skin\",\n      \"pmids\": [\"22829784\", \"22614019\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct biochemical contact between GRHL3 and Trithorax subunits not mapped\", \"Mechanism of miR-21 promoter repression not detailed\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Separated the insertase function of the Get1/2 transmembrane domains from cytosolic Get3 recruitment, and showed WRB+CAML are necessary and sufficient to constitute the receptor.\",\n      \"evidence\": \"Cell reporter assays and biochemical reconstitution with TM-domain mutants; yeast complementation by WRB/CAML with binding measurements\",\n      \"pmids\": [\"25043001\", \"24392163\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physical nature of the insertion conduit not yet defined\", \"Stoichiometry of the active complex unresolved\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Established the physiological importance of WRB-dependent TA insertion in excitable sensory cells, with otoferlin and synaptic-vesicle proteins as key clients and the TRC40-binding interface as functionally essential.\",\n      \"evidence\": \"Hair-cell and photoreceptor Wrb knockout/mutant in zebrafish and mice with electrophysiology, EM, immunostaining, and R73A binding-defective rescue\",\n      \"pmids\": [\"27458190\", \"27273592\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Full client repertoire in these cells not enumerated\", \"Whether vesicle defects are entirely TA-insertion-dependent not fully resolved\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Showed that in vitro client status does not predict in vivo receptor dependence, identifying syntaxin 5 as exquisitely WRB-sensitive and an autophagy target upon pathway failure.\",\n      \"evidence\": \"Tissue-specific WRB knockout mice with western blotting, TA-fate assays, and a yeast-to-mammalian client screen\",\n      \"pmids\": [\"28000760\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Determinants of differential substrate sensitivity unknown\", \"Single lab\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Revealed a Get3-independent role for the Get1/2 insertase in mitophagy, implicating a membrane-intrinsic function downstream of Atg32.\",\n      \"evidence\": \"Yeast get1/2 and get3 deletion mutants with mitophagy assays during respiratory growth\",\n      \"pmids\": [\"29673596\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"The relevant TA substrate(s) for mitophagy not identified\", \"Whether the role is conserved in mammals untested here\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Quantified how Get1/2 complex assembly cooperatively enhances Get3\\u2022TA binding, defining Get2 MoRFs and Get1 remodeling as jointly required for substrate release.\",\n      \"evidence\": \"Fluorescence binding assays, single-molecule FRET, and in vivo insertion assays with MoRF mutants\",\n      \"pmids\": [\"34614151\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Precise ordering of conformational steps during release not fully resolved\", \"Coupling to membrane insertion step not addressed\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Demonstrated that Get1/2 forms a gated aqueous channel whose opening, sealed by Get3, is required for TA release and possibly translocation of C-terminal hydrophilic segments.\",\n      \"evidence\": \"Bulk fluorescence and microfluidics channel assays in reconstituted bilayers with channel-activity mutants\",\n      \"pmids\": [\"36640319\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Channel model not yet independently replicated\", \"Direct evidence for C-terminal translocation not provided\", \"No high-resolution channel structure\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unresolved how the cytosolic substrate-release machinery is physically coupled to the transmembrane insertase/channel within a single Get1/2(WRB/CAML)\\u2013Get3 reaction cycle, and what governs substrate-specific receptor dependence in vivo.\",\n      \"evidence\": \"No single study in the timeline integrates the conformational opening, channel gating, and in vivo client-selectivity findings\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No integrated structure of the full membrane-embedded WRB/CAML\\u2013Get3\\u2013substrate intermediate\", \"Rules predicting which TA proteins require WRB in vivo unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [1, 5, 7]},\n      {\"term_id\": \"GO:0005198\", \"supporting_discovery_ids\": [1, 4, 7]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [12, 14, 15, 16]},\n      {\"term_id\": \"GO:0003677\", \"supporting_discovery_ids\": [12, 14]},\n      {\"term_id\": \"GO:0005215\", \"supporting_discovery_ids\": [7]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [0, 1, 2, 5]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0, 1, 17]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [12, 14]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [0, 1, 2]},\n      {\"term_id\": \"R-HSA-9609507\", \"supporting_discovery_ids\": [0, 1, 4]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [12, 14, 15, 16]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [13, 14]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [11]}\n    ],\n    \"complexes\": [\n      \"GET insertase complex (WRB/Get1\\u2013CAML/Get2)\"\n    ],\n    \"partners\": [\n      \"TRC40\",\n      \"Get3\",\n      \"CAML\",\n      \"LMO4\",\n      \"SLC4A1\",\n      \"GYPA\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":7,"faith_pct":85.71428571428571}}