{"gene":"EEF1D","run_date":"2026-06-09T23:54:42","timeline":{"discoveries":[{"year":2011,"finding":"EEF1D is a bona fide physiological substrate of protein kinase CK2, which directly phosphorylates EEF1D at serine 162 (S162). This was demonstrated by CK2 inhibitor-dependent decreases in EEF1D phosphorylation in 32P-labeled HeLa cells, direct CK2 kinase assays with FLAG-tagged EEF1D, λ-phosphatase treatment causing dramatic increases in phosphorylation, a phospho-specific antibody recognizing EEF1D pS162, and restoration of phosphorylation using CK2 inhibitor-resistant mutants.","method":"32P metabolic labeling, in vitro CK2 kinase assay, λ-phosphatase treatment, phospho-specific antibody (pS162), CK2 inhibitor-resistant mutants, 2D electrophoresis and mass spectrometry","journal":"Journal of proteome research","confidence":"High","confidence_rationale":"Tier 1 / Strong — multiple orthogonal methods including direct in vitro kinase assay, site-specific mutagenesis of kinase (inhibitor-resistant mutants), phospho-specific antibody, and metabolic labeling in a single rigorous study","pmids":["21936567"],"is_preprint":false},{"year":2015,"finding":"EEF1D undergoes alternative splicing to produce a long isoform and short isoforms with distinct functions: the long isoform acts as a transcriptional activator of heat-shock responsive genes (not as a translational elongation factor), while the short isoforms function in translation elongation. The long isoform regulates the cellular stress response through transcriptional activation.","method":"Alternative splicing characterization, functional analysis of isoforms in heat shock response and transcriptional activation (review summarizing experimental findings)","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — review summarizing prior experimental discoveries on isoform-specific functions; isoform distinction supported by molecular characterization but primary experimental details not fully described in this abstract","pmids":["25686034"],"is_preprint":false},{"year":2016,"finding":"EEF1D knockdown in oral squamous cell carcinoma (OSCC) cells reduced cell proliferation and induced epithelial-mesenchymal transition (EMT) phenotypes including cell invasion, while EEF1D and its interaction partners promote activation of cyclin D1 and vimentin proteins.","method":"siRNA knockdown, cell proliferation assay, invasion assay, Western blotting for cyclin D1 and vimentin, protein interaction partner identification","journal":"Clinical science (London, England : 1979)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function with defined cellular phenotype (proliferation, EMT/invasion) and specific molecular readouts (cyclin D1, vimentin), single lab","pmids":["26823560"],"is_preprint":false},{"year":2018,"finding":"EEF1D knockdown in osteosarcoma cells inhibited proliferation, colony-forming ability, and G2/M cell cycle transition, and decreased levels of phospho-Akt, phospho-mTOR, and phospho-Bad, placing EEF1D upstream of the Akt-mTOR and Akt-Bad signaling pathways.","method":"siRNA knockdown, cell proliferation assay, colony formation assay, cell cycle analysis, PathScan intracellular signaling array, Western blotting","journal":"Journal of experimental & clinical cancer research : CR","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function with specific signaling pathway readouts using multiple assays, single lab","pmids":["29510727"],"is_preprint":false},{"year":2019,"finding":"Biallelic loss-of-function variants in EEF1D that exclusively target the long isoform cause autosomal recessive intellectual disability, implicating the long isoform's heat shock response pathway (transcriptional activation function) rather than canonical translational elongation in the neurodevelopmental phenotype.","method":"SNP-based linkage analysis, whole exome sequencing, isoform-specific variant analysis in consanguineous family","journal":"Journal of human genetics","confidence":"Low","confidence_rationale":"Tier 3 / Weak — human genetic evidence from a single family; isoform specificity of pathogenic mechanism inferred from variant location, not directly demonstrated by functional experiment","pmids":["30787422"],"is_preprint":false},{"year":2021,"finding":"EEF1D regulates milk lipid synthesis in mammals: RNAi-mediated knockdown in primary bovine mammary epithelial cells caused aberrant lipid droplet formation and decreased milk triglyceride levels by 37.7%, acting via insulin (PI3K-Akt), AMPK, and PPAR pathways. CRISPR/Cas9 knockout mice showed incompletely developed mammary glands and decreased milk triglyceride by 23.4%.","method":"RNAi knockdown in primary bovine mammary epithelial cells, CRISPR/Cas9 knockout mice, lipid droplet imaging, triglyceride measurement, gene expression analysis of PI3K-Akt/AMPK/PPAR pathways","journal":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — two orthogonal loss-of-function approaches (RNAi and CRISPR KO) in two species with consistent phenotypic and pathway readouts","pmids":["33913197"],"is_preprint":false},{"year":2022,"finding":"EEF1D knockdown or knockout sensitizes ovarian cancer cells to cisplatin (DDP), partially via inactivation of the PI3K/AKT signaling pathway, leading to increased apoptosis (elevated cleaved caspase-3, altered Bcl-2/Bax) and decreased DNA damage repair (reduced ERCC1). EEF1D loss also affected OPTN levels.","method":"siRNA knockdown, CRISPR/Cas9 knockout, cell viability assay, apoptosis assay, xenograft mouse model, Western blotting for p-Akt, Bcl-2, Bax, cleaved caspase-3, ERCC1, OPTN","journal":"BMC cancer","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — two orthogonal loss-of-function approaches with in vivo validation and multiple molecular readouts, single lab","pmids":["35672728"],"is_preprint":false},{"year":2023,"finding":"The lncRNA NONMMUT033452.2 directly binds Eef1D (demonstrated by RNA pull-down assay), and overexpression of this lncRNA induces redistribution of Eef1D and substantially inhibits expression of downstream heat shock genes, revealing that Eef1D's heat shock gene regulation function can be sequestered by a lncRNA binding partner.","method":"RNA pull-down assay, FISH, cytoplasmic/nuclear fractionation, lncRNA overexpression, heat shock gene expression analysis","journal":"American journal of respiratory cell and molecular biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — RNA pull-down directly demonstrates binding; functional consequence on heat shock genes shown by overexpression; single lab","pmids":["36603194"],"is_preprint":false},{"year":2023,"finding":"Biallelic variants in the C-terminal GEF (guanine exchange factor) domain of EEF1D cause severe neurodevelopmental disorder with microcephaly and spasticity, indicating that the GEF domain—responsible for EEF1Bδ's role in catalyzing GTP/GDP exchange to reactivate eEF1A for aminoacyl-tRNA delivery—is essential for neurodevelopment, distinct from the alternatively spliced domain variants affecting the transcriptional function.","method":"Exome sequencing in two families, variant mapping to GEF domain, clinical phenotyping","journal":"Clinical genetics","confidence":"Low","confidence_rationale":"Tier 3 / Weak — human genetic evidence from two families; domain-based mechanistic inference without direct functional experiment on the GEF activity","pmids":["36576126"],"is_preprint":false},{"year":2024,"finding":"SRSF9 stabilizes EEF1D mRNA by binding to the 3'UTR of EEF1D mRNA (demonstrated by RNA immunoprecipitation and RNA pull-down assay), thereby upregulating EEF1D protein levels. EEF1D knockdown reversed the malignant proliferation and metastasis phenotype induced by SRSF9 overexpression in colorectal cancer cells.","method":"RNA immunoprecipitation, RNA pull-down assay, proteomics, EEF1D knockdown/overexpression, in vitro and in vivo proliferation/metastasis assays","journal":"International journal of cancer","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct RNA-protein interaction demonstrated by two orthogonal methods (RIP and RNA pull-down), epistasis shown by rescue experiment, single lab","pmids":["38771720"],"is_preprint":false},{"year":2024,"finding":"LGALS9B (Galectin-9B) binds EEF1D and competes with the E3 ubiquitin ligase HERC5 for this interaction, thereby preventing ubiquitin-proteasome-mediated degradation of EEF1D. The resulting EEF1D enrichment activates the PI3K/AKT signaling pathway to promote gastric cancer progression.","method":"Co-immunoprecipitation, competition binding assay between LGALS9B and HERC5 for EEF1D, proteasome inhibitor experiments, PI3K/AKT pathway readouts, in vitro and in vivo functional assays","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP demonstrates protein interactions; competitive binding and proteasomal degradation mechanism supported, single lab","pmids":["39639171"],"is_preprint":false},{"year":2025,"finding":"During Bovine alpha herpesvirus 1 (BoAHV1) productive infection, EEF1D undergoes enhanced nuclear translocation and forms puncta co-localizing with viral replication compartment marker ICP8 in the nucleus, while a portion of cytoplasmic EEF1D co-localizes with virion-associated protein gD. siRNA-mediated EEF1D knockdown significantly decreases BoAHV1 productive infection, indicating a proviral role for EEF1D.","method":"Immunofluorescence assay, siRNA knockdown, viral titer measurement, co-localization analysis","journal":"Veterinary microbiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct localization experiment with functional consequence (KD reduces viral infection); co-localization with viral proteins demonstrated, single lab","pmids":["40578269"],"is_preprint":false},{"year":2026,"finding":"KSHV RTA promotes EEF1D protein degradation via the ubiquitin-proteasome pathway and represses EEF1D transcription through promoter hypermethylation. RTA interacts with EEF1D protein and induces DNMT3A-dependent hypermethylation of the EEF1D promoter, a process facilitated by transcription factor PATZ1. EEF1D acts as an inhibitory factor for KSHV lytic reactivation, as EEF1D overexpression suppresses viral lytic replication while EEF1D depletion enhances viral reactivation.","method":"Co-immunoprecipitation (RTA-EEF1D interaction), dual-luciferase reporter assays, DNMT3A-dependent methylation assays, EEF1D overexpression/knockdown with viral reactivation readouts, promoter methylation analysis","journal":"Journal of virology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple complementary mechanisms demonstrated (protein interaction/degradation and transcriptional repression via methylation), with functional validation, single lab","pmids":["41805193"],"is_preprint":false},{"year":2026,"finding":"Short EEF1D isoforms containing exon 5 anchor the EEF1B complex to the endoplasmic reticulum (ER) by interacting with ER-resident scaffold proteins KTN1 and RRBP1. Deletion of exon 5 disrupts ER anchoring and causes diffuse cytoplasmic localization of the EEF1B complex, accompanied by reduced EEF1B subunit abundance in multiple tissues in vivo, without affecting global protein synthesis rates.","method":"FLAG-tagged EEF1D pulldown with mass spectrometry, exon 5 deletion mutant, exon 5 KO mice, subcellular fractionation/localization, protein abundance measurement across tissues, global protein synthesis rate assay","journal":"Life science alliance","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — mass spectrometry identifies interactors, deletion mutant confirms domain requirement, in vivo KO mouse validation with multiple tissue readouts, multiple orthogonal methods in single study","pmids":["42230146"],"is_preprint":false}],"current_model":"EEF1D encodes the eEF1Bδ subunit of the eEF1B guanine nucleotide exchange complex, which reactivates eEF1A for aminoacyl-tRNA delivery to the ribosome during translation elongation; its short isoforms (containing exon 5) anchor the EEF1B complex to the ER via KTN1/RRBP1 interactions, while the alternatively spliced long isoform functions as a transcriptional activator of heat-shock genes in the nucleus. EEF1D is directly phosphorylated at S162 by CK2, promotes cell proliferation and survival via PI3K/Akt-mTOR signaling, is stabilized post-translationally by LGALS9B competing with E3 ligase HERC5, and its mRNA is stabilized by SRSF9 binding to its 3'UTR. Loss-of-function variants in either the alternatively spliced domain (long isoform) or the GEF domain cause severe autosomal recessive neurodevelopmental disorders, and EEF1D plays functional roles in mammary gland lipid synthesis and viral replication."},"narrative":{"mechanistic_narrative":"EEF1D encodes eEF1Bδ, a subunit of the eEF1B guanine-nucleotide exchange complex that operates at the interface of translation elongation, the cellular stress response, and growth signaling [PMID:25686034, PMID:36576126]. Alternative splicing generates functionally distinct isoforms: short isoforms participate in translation elongation, whereas the alternatively spliced long isoform functions as a transcriptional activator of heat-shock responsive genes rather than an elongation factor [PMID:25686034]. Short isoforms containing exon 5 anchor the EEF1B complex to the endoplasmic reticulum through interactions with the ER scaffold proteins KTN1 and RRBP1; deletion of exon 5 causes diffuse cytoplasmic redistribution of the complex and reduced EEF1B subunit abundance in vivo without changing global protein synthesis rates [PMID:42230146]. The heat-shock transcriptional activity can be antagonized by direct binding of a lncRNA, which redistributes EEF1D and represses downstream heat-shock genes [PMID:36603194]. EEF1D is phosphorylated at serine 162 by protein kinase CK2 [PMID:21936567], and its abundance is controlled post-translationally — galectin LGALS9B competes with the E3 ligase HERC5 to block ubiquitin-proteasome degradation [PMID:39639171] — and post-transcriptionally by SRSF9 binding to its 3'UTR to stabilize the mRNA [PMID:38771720]. Across multiple cancers EEF1D promotes proliferation, survival, and metastasis largely through activation of PI3K/Akt-mTOR signaling, and its loss sensitizes cells to chemotherapy [PMID:29510727, PMID:35672728, PMID:39639171]. Biallelic loss-of-function variants targeting either the long-isoform/alternatively spliced domain or the C-terminal GEF domain cause severe autosomal recessive neurodevelopmental disorders [PMID:30787422, PMID:36576126]. EEF1D also supports mammary milk lipid synthesis [PMID:33913197] and is exploited by viruses as a host factor [PMID:40578269, PMID:41805193].","teleology":[{"year":2011,"claim":"Established that EEF1D is a direct, physiological substrate of CK2, defining a specific regulatory phosphorylation site (S162) on the elongation-factor subunit.","evidence":"32P metabolic labeling, in vitro CK2 kinase assays, λ-phosphatase treatment, phospho-specific pS162 antibody and inhibitor-resistant CK2 mutants in HeLa cells","pmids":["21936567"],"confidence":"High","gaps":["Functional consequence of S162 phosphorylation on GEF activity or complex assembly not resolved","Does not link phosphorylation to isoform-specific roles"]},{"year":2015,"claim":"Resolved that EEF1D isoforms have divergent functions, separating a translation-elongation role for short isoforms from a heat-shock transcriptional activation role for the long isoform.","evidence":"Review synthesizing alternative-splicing and isoform functional characterization in heat-shock response","pmids":["25686034"],"confidence":"Medium","gaps":["Primary experimental detail of transcriptional activation mechanism not described","Target promoters and co-factors of the long isoform unspecified"]},{"year":2016,"claim":"Connected EEF1D to oncogenic proliferation and EMT, moving it beyond a housekeeping translation role into cancer cell biology.","evidence":"siRNA knockdown in oral squamous carcinoma cells with proliferation/invasion assays and cyclin D1/vimentin readouts","pmids":["26823560"],"confidence":"Medium","gaps":["Which isoform drives the phenotype not distinguished","Direct molecular link between EEF1D and cyclin D1/vimentin not defined"]},{"year":2018,"claim":"Placed EEF1D upstream of Akt-mTOR/Akt-Bad signaling, identifying the pathway through which it drives proliferation and survival.","evidence":"siRNA knockdown in osteosarcoma cells with cell cycle analysis, signaling array, and phospho-Akt/mTOR/Bad Western blots","pmids":["29510727"],"confidence":"Medium","gaps":["Mechanism by which EEF1D activates Akt not defined","Direct vs indirect effect on signaling unresolved"]},{"year":2019,"claim":"Implicated the long isoform / heat-shock transcriptional pathway, rather than canonical elongation, in a human neurodevelopmental disorder.","evidence":"Linkage analysis and whole-exome sequencing of a consanguineous family with isoform-specific variant mapping","pmids":["30787422"],"confidence":"Low","gaps":["Single family; isoform-specific pathomechanism inferred from variant location, not demonstrated functionally","No rescue or animal model"]},{"year":2021,"claim":"Demonstrated a physiological role for EEF1D in mammary lipid metabolism, broadening its function to tissue-specific metabolic output.","evidence":"RNAi in bovine mammary epithelial cells and CRISPR/Cas9 knockout mice with lipid droplet imaging, triglyceride measurement and PI3K-Akt/AMPK/PPAR pathway readouts","pmids":["33913197"],"confidence":"High","gaps":["Whether effect is mediated by translation or signaling function unclear","Isoform responsible not identified"]},{"year":2022,"claim":"Showed EEF1D loss sensitizes cancer cells to chemotherapy via PI3K/AKT inactivation, linking it to apoptosis and DNA-damage-repair regulation.","evidence":"siRNA and CRISPR knockout in ovarian cancer cells with apoptosis assays, xenograft model, and Western blots for p-Akt, Bcl-2/Bax, cleaved caspase-3, ERCC1, OPTN","pmids":["35672728"],"confidence":"Medium","gaps":["Direct effect of EEF1D on ERCC1/OPTN not mechanistically defined","Single lab"]},{"year":2023,"claim":"Revealed that EEF1D's heat-shock transcriptional function is regulated by lncRNA sequestration, providing a mechanism for controlling its nuclear activity.","evidence":"RNA pull-down, FISH, nuclear/cytoplasmic fractionation, and lncRNA overexpression with heat-shock gene readouts","pmids":["36603194"],"confidence":"Medium","gaps":["Human ortholog lncRNA equivalence not established","Binding region on EEF1D not mapped"]},{"year":2023,"claim":"Identified the C-terminal GEF domain as essential for neurodevelopment, establishing a second, distinct disease mechanism from the alternatively spliced domain.","evidence":"Exome sequencing in two families with variant mapping to the GEF domain and clinical phenotyping","pmids":["36576126"],"confidence":"Low","gaps":["Effect on GEF/eEF1A reactivation activity not functionally tested","No animal or cellular model"]},{"year":2024,"claim":"Defined post-transcriptional control of EEF1D, showing SRSF9 stabilizes its mRNA to drive cancer proliferation and metastasis.","evidence":"RNA immunoprecipitation, RNA pull-down, proteomics, and EEF1D knockdown rescue of SRSF9-driven phenotypes in colorectal cancer cells","pmids":["38771720"],"confidence":"Medium","gaps":["3'UTR binding site not mapped","Whether stabilization is isoform-selective unknown"]},{"year":2024,"claim":"Defined post-translational control of EEF1D abundance through a competition between LGALS9B and the E3 ligase HERC5, linking stability directly to PI3K/AKT-driven tumor progression.","evidence":"Co-IP, competition binding assays, proteasome inhibitor experiments and in vitro/in vivo functional assays in gastric cancer","pmids":["39639171"],"confidence":"Medium","gaps":["Ubiquitination site on EEF1D not mapped","Reciprocal validation in additional models lacking"]},{"year":2025,"claim":"Identified EEF1D as a proviral host factor that translocates to viral replication compartments during herpesvirus infection.","evidence":"Immunofluorescence co-localization with ICP8 and gD, siRNA knockdown, and viral titer measurement during BoAHV1 infection","pmids":["40578269"],"confidence":"Medium","gaps":["Molecular function of EEF1D within replication compartments unknown","Direct viral protein binding not demonstrated"]},{"year":2026,"claim":"Showed that KSHV RTA suppresses EEF1D both by proteasomal degradation and promoter hypermethylation, establishing EEF1D as a host restriction factor for viral lytic reactivation.","evidence":"Co-IP, dual-luciferase reporters, DNMT3A-dependent methylation assays and EEF1D overexpression/knockdown with viral reactivation readouts","pmids":["41805193"],"confidence":"Medium","gaps":["Mechanism by which EEF1D restricts lytic replication not defined","Single lab"]},{"year":2026,"claim":"Established the molecular basis of EEF1B complex localization, showing exon-5-containing short isoforms anchor the complex to the ER via KTN1/RRBP1 independent of bulk translation.","evidence":"FLAG-EEF1D pulldown/MS, exon 5 deletion mutant, exon 5 KO mice, subcellular fractionation, multi-tissue abundance and global protein synthesis assays","pmids":["42230146"],"confidence":"High","gaps":["Functional purpose of ER anchoring (localized translation?) not established","Connection between ER anchoring and disease/cancer phenotypes untested"]},{"year":null,"claim":"How EEF1D's distinct biochemical activities — GEF-mediated elongation, ER anchoring, and long-isoform heat-shock transcription — are coordinated, and which activity underlies its disease, metabolic, and cancer roles, remains unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["Isoform attribution of most cancer/metabolic phenotypes not defined","No structural model linking GEF and transcriptional functions","Direct demonstration of GEF activity loss in disease variants lacking"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[1,7]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[8]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[13]}],"localization":[{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[13]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[7,11]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[11,13]}],"pathway":[{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[8,13]},{"term_id":"R-HSA-8953897","term_label":"Cellular responses to stimuli","supporting_discovery_ids":[1,7]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[3,10]}],"complexes":["eEF1B guanine nucleotide exchange complex"],"partners":["KTN1","RRBP1","LGALS9B","HERC5","SRSF9","CK2"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P29692","full_name":"Elongation factor 1-delta","aliases":["Antigen NY-CO-4"],"length_aa":281,"mass_kda":31.1,"function":"EF-1-beta and EF-1-delta stimulate the exchange of GDP bound to EF-1-alpha to GTP, regenerating EF-1-alpha for another round of transfer of aminoacyl-tRNAs to the ribosome Regulates induction of heat-shock-responsive genes through association with heat shock transcription factors and direct DNA-binding at heat shock promoter elements (HSE)","subcellular_location":"Nucleus","url":"https://www.uniprot.org/uniprotkb/P29692/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/EEF1D","classification":"Not Classified","n_dependent_lines":34,"n_total_lines":1208,"dependency_fraction":0.028145695364238412},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"EEF1G","stoichiometry":10.0}],"url":"https://opencell.sf.czbiohub.org/search/EEF1D","total_profiled":1310},"omim":[{"mim_id":"621150","title":"NEURODEVELOPMENTAL DISORDER WITH THIN CORPUS CALLOSUM, HYPOTONIA, AND ABSENT LANGUAGE; NEDTCHAL","url":"https://www.omim.org/entry/621150"},{"mim_id":"600655","title":"EUKARYOTIC TRANSLATION ELONGATION FACTOR 1, BETA-2; EEF1B2","url":"https://www.omim.org/entry/600655"},{"mim_id":"600381","title":"KINECTIN; KTN1","url":"https://www.omim.org/entry/600381"},{"mim_id":"153550","title":"CHROMOSOME 5q DELETION SYNDROME","url":"https://www.omim.org/entry/153550"},{"mim_id":"130593","title":"EUKARYOTIC TRANSLATION ELONGATION FACTOR 1, GAMMA; EEF1G","url":"https://www.omim.org/entry/130593"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nucleoplasm","reliability":"Supported"},{"location":"Nucleoli fibrillar center","reliability":"Supported"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/EEF1D"},"hgnc":{"alias_symbol":["EF-1D","FLJ20897"],"prev_symbol":[]},"alphafold":{"accession":"P29692","domains":[{"cath_id":"3.30.70.60","chopping":"203-281","consensus_level":"high","plddt":83.2876,"start":203,"end":281}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P29692","model_url":"https://alphafold.ebi.ac.uk/files/AF-P29692-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P29692-F1-predicted_aligned_error_v6.png","plddt_mean":73.0},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=EEF1D","jax_strain_url":"https://www.jax.org/strain/search?query=EEF1D"},"sequence":{"accession":"P29692","fasta_url":"https://rest.uniprot.org/uniprotkb/P29692.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P29692/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P29692"}},"corpus_meta":[{"pmid":"16968546","id":"PMC_16968546","title":"Medulloblastoma outcome is adversely associated with overexpression of EEF1D, RPL30, and RPS20 on the long arm of chromosome 8.","date":"2006","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/16968546","citation_count":62,"is_preprint":false},{"pmid":"26823560","id":"PMC_26823560","title":"EEF1D modulates proliferation and epithelial-mesenchymal transition in oral squamous cell carcinoma.","date":"2016","source":"Clinical science (London, England : 1979)","url":"https://pubmed.ncbi.nlm.nih.gov/26823560","citation_count":34,"is_preprint":false},{"pmid":"29510727","id":"PMC_29510727","title":"EEF1D overexpression promotes osteosarcoma cell proliferation by facilitating Akt-mTOR and Akt-bad signaling.","date":"2018","source":"Journal of experimental & clinical cancer research : CR","url":"https://pubmed.ncbi.nlm.nih.gov/29510727","citation_count":29,"is_preprint":false},{"pmid":"21936567","id":"PMC_21936567","title":"Unbiased functional proteomics strategy for protein kinase inhibitor validation and identification of bona fide protein kinase substrates: application to identification of EEF1D as a substrate for CK2.","date":"2011","source":"Journal of proteome research","url":"https://pubmed.ncbi.nlm.nih.gov/21936567","citation_count":26,"is_preprint":false},{"pmid":"33029523","id":"PMC_33029523","title":"EEF1D Promotes Glioma Proliferation, Migration, and Invasion through EMT and PI3K/Akt Pathway.","date":"2020","source":"BioMed research international","url":"https://pubmed.ncbi.nlm.nih.gov/33029523","citation_count":23,"is_preprint":false},{"pmid":"25686034","id":"PMC_25686034","title":"Regulation of translation factor EEF1D gene function by alternative splicing.","date":"2015","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/25686034","citation_count":17,"is_preprint":false},{"pmid":"33913197","id":"PMC_33913197","title":"EEF1D facilitates milk lipid synthesis by regulation of PI3K-Akt signaling in mammals.","date":"2021","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/33913197","citation_count":17,"is_preprint":false},{"pmid":"28667419","id":"PMC_28667419","title":"Regulation of DNA methylation on EEF1D and RPL8 expression in cattle.","date":"2017","source":"Genetica","url":"https://pubmed.ncbi.nlm.nih.gov/28667419","citation_count":16,"is_preprint":false},{"pmid":"30787422","id":"PMC_30787422","title":"Biallelic loss of EEF1D function links heat shock response pathway to autosomal recessive intellectual disability.","date":"2019","source":"Journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/30787422","citation_count":13,"is_preprint":false},{"pmid":"24239553","id":"PMC_24239553","title":"Identification and expression pattern of two novel alternative splicing variants of EEF1D gene of dairy cattle.","date":"2013","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/24239553","citation_count":8,"is_preprint":false},{"pmid":"34790806","id":"PMC_34790806","title":"The role of EEF1D in disease pathogenesis: a narrative review.","date":"2021","source":"Annals of translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/34790806","citation_count":7,"is_preprint":false},{"pmid":"36603194","id":"PMC_36603194","title":"Prenatal LPS Exposure Promotes Allergic Airway Inflammation via Long Coding RNA NONMMUT033452.2, and Protein Binding Partner, Eef1D.","date":"2023","source":"American journal of respiratory cell and molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/36603194","citation_count":7,"is_preprint":false},{"pmid":"35672728","id":"PMC_35672728","title":"Interfering with the expression of EEF1D gene enhances the sensitivity of ovarian cancer cells to cisplatin.","date":"2022","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/35672728","citation_count":6,"is_preprint":false},{"pmid":"38771720","id":"PMC_38771720","title":"EEF1D stabilized by SRSF9 promotes colorectal cancer via enhancing the proliferation and metastasis.","date":"2024","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/38771720","citation_count":4,"is_preprint":false},{"pmid":"26794801","id":"PMC_26794801","title":"Identification of novel isoforms of dairy goat EEF1D and their mRNA expression characterization.","date":"2016","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/26794801","citation_count":4,"is_preprint":false},{"pmid":"36576126","id":"PMC_36576126","title":"Expanding the spectrum of EEF1D neurodevelopmental disorders: Biallelic variants in the guanine exchange domain.","date":"2023","source":"Clinical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/36576126","citation_count":3,"is_preprint":false},{"pmid":"39639171","id":"PMC_39639171","title":"LGALS9B stabilizes EEF1D protein and activates the PI3K/AKT signaling pathway to promote gastric cancer occurrence and metastasis.","date":"2024","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/39639171","citation_count":2,"is_preprint":false},{"pmid":"40578269","id":"PMC_40578269","title":"EEF1D signaling contributes to Bovine alpha herpesvirus 1 productive infection, potentially through regulation of viral replication compartments.","date":"2025","source":"Veterinary microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/40578269","citation_count":1,"is_preprint":false},{"pmid":"38083972","id":"PMC_38083972","title":"Autosomal recessive intellectual disability caused by compound heterozygous variants of the EEF1D gene in a Chinese family.","date":"2023","source":"Molecular genetics & genomic medicine","url":"https://pubmed.ncbi.nlm.nih.gov/38083972","citation_count":0,"is_preprint":false},{"pmid":"41805193","id":"PMC_41805193","title":"Repression of EEF1D by KSHV RTA promotes viral lytic reactivation.","date":"2026","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/41805193","citation_count":0,"is_preprint":false},{"pmid":"42230146","id":"PMC_42230146","title":"The ER anchoring and abundance of the EEF1B complex is affected by tissue-specific alternative EEF1D splicing.","date":"2026","source":"Life science alliance","url":"https://pubmed.ncbi.nlm.nih.gov/42230146","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":12052,"output_tokens":4007,"usd":0.04813,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11860,"output_tokens":4421,"usd":0.084913,"stage2_stop_reason":"end_turn"},"total_usd":0.133043,"stage1_batch_id":"msgbatch_01AsR7g8dGTEiEw9U83bJeWn","stage2_batch_id":"msgbatch_01YV5ZyxtFNUZEjxRcpd9jzb","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2011,\n      \"finding\": \"EEF1D is a bona fide physiological substrate of protein kinase CK2, which directly phosphorylates EEF1D at serine 162 (S162). This was demonstrated by CK2 inhibitor-dependent decreases in EEF1D phosphorylation in 32P-labeled HeLa cells, direct CK2 kinase assays with FLAG-tagged EEF1D, λ-phosphatase treatment causing dramatic increases in phosphorylation, a phospho-specific antibody recognizing EEF1D pS162, and restoration of phosphorylation using CK2 inhibitor-resistant mutants.\",\n      \"method\": \"32P metabolic labeling, in vitro CK2 kinase assay, λ-phosphatase treatment, phospho-specific antibody (pS162), CK2 inhibitor-resistant mutants, 2D electrophoresis and mass spectrometry\",\n      \"journal\": \"Journal of proteome research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — multiple orthogonal methods including direct in vitro kinase assay, site-specific mutagenesis of kinase (inhibitor-resistant mutants), phospho-specific antibody, and metabolic labeling in a single rigorous study\",\n      \"pmids\": [\"21936567\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"EEF1D undergoes alternative splicing to produce a long isoform and short isoforms with distinct functions: the long isoform acts as a transcriptional activator of heat-shock responsive genes (not as a translational elongation factor), while the short isoforms function in translation elongation. The long isoform regulates the cellular stress response through transcriptional activation.\",\n      \"method\": \"Alternative splicing characterization, functional analysis of isoforms in heat shock response and transcriptional activation (review summarizing experimental findings)\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — review summarizing prior experimental discoveries on isoform-specific functions; isoform distinction supported by molecular characterization but primary experimental details not fully described in this abstract\",\n      \"pmids\": [\"25686034\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"EEF1D knockdown in oral squamous cell carcinoma (OSCC) cells reduced cell proliferation and induced epithelial-mesenchymal transition (EMT) phenotypes including cell invasion, while EEF1D and its interaction partners promote activation of cyclin D1 and vimentin proteins.\",\n      \"method\": \"siRNA knockdown, cell proliferation assay, invasion assay, Western blotting for cyclin D1 and vimentin, protein interaction partner identification\",\n      \"journal\": \"Clinical science (London, England : 1979)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with defined cellular phenotype (proliferation, EMT/invasion) and specific molecular readouts (cyclin D1, vimentin), single lab\",\n      \"pmids\": [\"26823560\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"EEF1D knockdown in osteosarcoma cells inhibited proliferation, colony-forming ability, and G2/M cell cycle transition, and decreased levels of phospho-Akt, phospho-mTOR, and phospho-Bad, placing EEF1D upstream of the Akt-mTOR and Akt-Bad signaling pathways.\",\n      \"method\": \"siRNA knockdown, cell proliferation assay, colony formation assay, cell cycle analysis, PathScan intracellular signaling array, Western blotting\",\n      \"journal\": \"Journal of experimental & clinical cancer research : CR\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with specific signaling pathway readouts using multiple assays, single lab\",\n      \"pmids\": [\"29510727\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Biallelic loss-of-function variants in EEF1D that exclusively target the long isoform cause autosomal recessive intellectual disability, implicating the long isoform's heat shock response pathway (transcriptional activation function) rather than canonical translational elongation in the neurodevelopmental phenotype.\",\n      \"method\": \"SNP-based linkage analysis, whole exome sequencing, isoform-specific variant analysis in consanguineous family\",\n      \"journal\": \"Journal of human genetics\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — human genetic evidence from a single family; isoform specificity of pathogenic mechanism inferred from variant location, not directly demonstrated by functional experiment\",\n      \"pmids\": [\"30787422\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"EEF1D regulates milk lipid synthesis in mammals: RNAi-mediated knockdown in primary bovine mammary epithelial cells caused aberrant lipid droplet formation and decreased milk triglyceride levels by 37.7%, acting via insulin (PI3K-Akt), AMPK, and PPAR pathways. CRISPR/Cas9 knockout mice showed incompletely developed mammary glands and decreased milk triglyceride by 23.4%.\",\n      \"method\": \"RNAi knockdown in primary bovine mammary epithelial cells, CRISPR/Cas9 knockout mice, lipid droplet imaging, triglyceride measurement, gene expression analysis of PI3K-Akt/AMPK/PPAR pathways\",\n      \"journal\": \"FASEB journal : official publication of the Federation of American Societies for Experimental Biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — two orthogonal loss-of-function approaches (RNAi and CRISPR KO) in two species with consistent phenotypic and pathway readouts\",\n      \"pmids\": [\"33913197\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"EEF1D knockdown or knockout sensitizes ovarian cancer cells to cisplatin (DDP), partially via inactivation of the PI3K/AKT signaling pathway, leading to increased apoptosis (elevated cleaved caspase-3, altered Bcl-2/Bax) and decreased DNA damage repair (reduced ERCC1). EEF1D loss also affected OPTN levels.\",\n      \"method\": \"siRNA knockdown, CRISPR/Cas9 knockout, cell viability assay, apoptosis assay, xenograft mouse model, Western blotting for p-Akt, Bcl-2, Bax, cleaved caspase-3, ERCC1, OPTN\",\n      \"journal\": \"BMC cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — two orthogonal loss-of-function approaches with in vivo validation and multiple molecular readouts, single lab\",\n      \"pmids\": [\"35672728\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"The lncRNA NONMMUT033452.2 directly binds Eef1D (demonstrated by RNA pull-down assay), and overexpression of this lncRNA induces redistribution of Eef1D and substantially inhibits expression of downstream heat shock genes, revealing that Eef1D's heat shock gene regulation function can be sequestered by a lncRNA binding partner.\",\n      \"method\": \"RNA pull-down assay, FISH, cytoplasmic/nuclear fractionation, lncRNA overexpression, heat shock gene expression analysis\",\n      \"journal\": \"American journal of respiratory cell and molecular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — RNA pull-down directly demonstrates binding; functional consequence on heat shock genes shown by overexpression; single lab\",\n      \"pmids\": [\"36603194\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Biallelic variants in the C-terminal GEF (guanine exchange factor) domain of EEF1D cause severe neurodevelopmental disorder with microcephaly and spasticity, indicating that the GEF domain—responsible for EEF1Bδ's role in catalyzing GTP/GDP exchange to reactivate eEF1A for aminoacyl-tRNA delivery—is essential for neurodevelopment, distinct from the alternatively spliced domain variants affecting the transcriptional function.\",\n      \"method\": \"Exome sequencing in two families, variant mapping to GEF domain, clinical phenotyping\",\n      \"journal\": \"Clinical genetics\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — human genetic evidence from two families; domain-based mechanistic inference without direct functional experiment on the GEF activity\",\n      \"pmids\": [\"36576126\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"SRSF9 stabilizes EEF1D mRNA by binding to the 3'UTR of EEF1D mRNA (demonstrated by RNA immunoprecipitation and RNA pull-down assay), thereby upregulating EEF1D protein levels. EEF1D knockdown reversed the malignant proliferation and metastasis phenotype induced by SRSF9 overexpression in colorectal cancer cells.\",\n      \"method\": \"RNA immunoprecipitation, RNA pull-down assay, proteomics, EEF1D knockdown/overexpression, in vitro and in vivo proliferation/metastasis assays\",\n      \"journal\": \"International journal of cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct RNA-protein interaction demonstrated by two orthogonal methods (RIP and RNA pull-down), epistasis shown by rescue experiment, single lab\",\n      \"pmids\": [\"38771720\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"LGALS9B (Galectin-9B) binds EEF1D and competes with the E3 ubiquitin ligase HERC5 for this interaction, thereby preventing ubiquitin-proteasome-mediated degradation of EEF1D. The resulting EEF1D enrichment activates the PI3K/AKT signaling pathway to promote gastric cancer progression.\",\n      \"method\": \"Co-immunoprecipitation, competition binding assay between LGALS9B and HERC5 for EEF1D, proteasome inhibitor experiments, PI3K/AKT pathway readouts, in vitro and in vivo functional assays\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP demonstrates protein interactions; competitive binding and proteasomal degradation mechanism supported, single lab\",\n      \"pmids\": [\"39639171\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"During Bovine alpha herpesvirus 1 (BoAHV1) productive infection, EEF1D undergoes enhanced nuclear translocation and forms puncta co-localizing with viral replication compartment marker ICP8 in the nucleus, while a portion of cytoplasmic EEF1D co-localizes with virion-associated protein gD. siRNA-mediated EEF1D knockdown significantly decreases BoAHV1 productive infection, indicating a proviral role for EEF1D.\",\n      \"method\": \"Immunofluorescence assay, siRNA knockdown, viral titer measurement, co-localization analysis\",\n      \"journal\": \"Veterinary microbiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct localization experiment with functional consequence (KD reduces viral infection); co-localization with viral proteins demonstrated, single lab\",\n      \"pmids\": [\"40578269\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"KSHV RTA promotes EEF1D protein degradation via the ubiquitin-proteasome pathway and represses EEF1D transcription through promoter hypermethylation. RTA interacts with EEF1D protein and induces DNMT3A-dependent hypermethylation of the EEF1D promoter, a process facilitated by transcription factor PATZ1. EEF1D acts as an inhibitory factor for KSHV lytic reactivation, as EEF1D overexpression suppresses viral lytic replication while EEF1D depletion enhances viral reactivation.\",\n      \"method\": \"Co-immunoprecipitation (RTA-EEF1D interaction), dual-luciferase reporter assays, DNMT3A-dependent methylation assays, EEF1D overexpression/knockdown with viral reactivation readouts, promoter methylation analysis\",\n      \"journal\": \"Journal of virology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple complementary mechanisms demonstrated (protein interaction/degradation and transcriptional repression via methylation), with functional validation, single lab\",\n      \"pmids\": [\"41805193\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"Short EEF1D isoforms containing exon 5 anchor the EEF1B complex to the endoplasmic reticulum (ER) by interacting with ER-resident scaffold proteins KTN1 and RRBP1. Deletion of exon 5 disrupts ER anchoring and causes diffuse cytoplasmic localization of the EEF1B complex, accompanied by reduced EEF1B subunit abundance in multiple tissues in vivo, without affecting global protein synthesis rates.\",\n      \"method\": \"FLAG-tagged EEF1D pulldown with mass spectrometry, exon 5 deletion mutant, exon 5 KO mice, subcellular fractionation/localization, protein abundance measurement across tissues, global protein synthesis rate assay\",\n      \"journal\": \"Life science alliance\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — mass spectrometry identifies interactors, deletion mutant confirms domain requirement, in vivo KO mouse validation with multiple tissue readouts, multiple orthogonal methods in single study\",\n      \"pmids\": [\"42230146\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"EEF1D encodes the eEF1Bδ subunit of the eEF1B guanine nucleotide exchange complex, which reactivates eEF1A for aminoacyl-tRNA delivery to the ribosome during translation elongation; its short isoforms (containing exon 5) anchor the EEF1B complex to the ER via KTN1/RRBP1 interactions, while the alternatively spliced long isoform functions as a transcriptional activator of heat-shock genes in the nucleus. EEF1D is directly phosphorylated at S162 by CK2, promotes cell proliferation and survival via PI3K/Akt-mTOR signaling, is stabilized post-translationally by LGALS9B competing with E3 ligase HERC5, and its mRNA is stabilized by SRSF9 binding to its 3'UTR. Loss-of-function variants in either the alternatively spliced domain (long isoform) or the GEF domain cause severe autosomal recessive neurodevelopmental disorders, and EEF1D plays functional roles in mammary gland lipid synthesis and viral replication.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"EEF1D encodes eEF1Bδ, a subunit of the eEF1B guanine-nucleotide exchange complex that operates at the interface of translation elongation, the cellular stress response, and growth signaling [#1, #8]. Alternative splicing generates functionally distinct isoforms: short isoforms participate in translation elongation, whereas the alternatively spliced long isoform functions as a transcriptional activator of heat-shock responsive genes rather than an elongation factor [#1]. Short isoforms containing exon 5 anchor the EEF1B complex to the endoplasmic reticulum through interactions with the ER scaffold proteins KTN1 and RRBP1; deletion of exon 5 causes diffuse cytoplasmic redistribution of the complex and reduced EEF1B subunit abundance in vivo without changing global protein synthesis rates [#13]. The heat-shock transcriptional activity can be antagonized by direct binding of a lncRNA, which redistributes EEF1D and represses downstream heat-shock genes [#7]. EEF1D is phosphorylated at serine 162 by protein kinase CK2 [#0], and its abundance is controlled post-translationally — galectin LGALS9B competes with the E3 ligase HERC5 to block ubiquitin-proteasome degradation [#10] — and post-transcriptionally by SRSF9 binding to its 3'UTR to stabilize the mRNA [#9]. Across multiple cancers EEF1D promotes proliferation, survival, and metastasis largely through activation of PI3K/Akt-mTOR signaling, and its loss sensitizes cells to chemotherapy [#3, #6, #10]. Biallelic loss-of-function variants targeting either the long-isoform/alternatively spliced domain or the C-terminal GEF domain cause severe autosomal recessive neurodevelopmental disorders [#4, #8]. EEF1D also supports mammary milk lipid synthesis [#5] and is exploited by viruses as a host factor [#11, #12].\",\n  \"teleology\": [\n    {\n      \"year\": 2011,\n      \"claim\": \"Established that EEF1D is a direct, physiological substrate of CK2, defining a specific regulatory phosphorylation site (S162) on the elongation-factor subunit.\",\n      \"evidence\": \"32P metabolic labeling, in vitro CK2 kinase assays, λ-phosphatase treatment, phospho-specific pS162 antibody and inhibitor-resistant CK2 mutants in HeLa cells\",\n      \"pmids\": [\"21936567\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of S162 phosphorylation on GEF activity or complex assembly not resolved\", \"Does not link phosphorylation to isoform-specific roles\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Resolved that EEF1D isoforms have divergent functions, separating a translation-elongation role for short isoforms from a heat-shock transcriptional activation role for the long isoform.\",\n      \"evidence\": \"Review synthesizing alternative-splicing and isoform functional characterization in heat-shock response\",\n      \"pmids\": [\"25686034\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Primary experimental detail of transcriptional activation mechanism not described\", \"Target promoters and co-factors of the long isoform unspecified\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Connected EEF1D to oncogenic proliferation and EMT, moving it beyond a housekeeping translation role into cancer cell biology.\",\n      \"evidence\": \"siRNA knockdown in oral squamous carcinoma cells with proliferation/invasion assays and cyclin D1/vimentin readouts\",\n      \"pmids\": [\"26823560\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Which isoform drives the phenotype not distinguished\", \"Direct molecular link between EEF1D and cyclin D1/vimentin not defined\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Placed EEF1D upstream of Akt-mTOR/Akt-Bad signaling, identifying the pathway through which it drives proliferation and survival.\",\n      \"evidence\": \"siRNA knockdown in osteosarcoma cells with cell cycle analysis, signaling array, and phospho-Akt/mTOR/Bad Western blots\",\n      \"pmids\": [\"29510727\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which EEF1D activates Akt not defined\", \"Direct vs indirect effect on signaling unresolved\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Implicated the long isoform / heat-shock transcriptional pathway, rather than canonical elongation, in a human neurodevelopmental disorder.\",\n      \"evidence\": \"Linkage analysis and whole-exome sequencing of a consanguineous family with isoform-specific variant mapping\",\n      \"pmids\": [\"30787422\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Single family; isoform-specific pathomechanism inferred from variant location, not demonstrated functionally\", \"No rescue or animal model\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Demonstrated a physiological role for EEF1D in mammary lipid metabolism, broadening its function to tissue-specific metabolic output.\",\n      \"evidence\": \"RNAi in bovine mammary epithelial cells and CRISPR/Cas9 knockout mice with lipid droplet imaging, triglyceride measurement and PI3K-Akt/AMPK/PPAR pathway readouts\",\n      \"pmids\": [\"33913197\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether effect is mediated by translation or signaling function unclear\", \"Isoform responsible not identified\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Showed EEF1D loss sensitizes cancer cells to chemotherapy via PI3K/AKT inactivation, linking it to apoptosis and DNA-damage-repair regulation.\",\n      \"evidence\": \"siRNA and CRISPR knockout in ovarian cancer cells with apoptosis assays, xenograft model, and Western blots for p-Akt, Bcl-2/Bax, cleaved caspase-3, ERCC1, OPTN\",\n      \"pmids\": [\"35672728\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct effect of EEF1D on ERCC1/OPTN not mechanistically defined\", \"Single lab\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Revealed that EEF1D's heat-shock transcriptional function is regulated by lncRNA sequestration, providing a mechanism for controlling its nuclear activity.\",\n      \"evidence\": \"RNA pull-down, FISH, nuclear/cytoplasmic fractionation, and lncRNA overexpression with heat-shock gene readouts\",\n      \"pmids\": [\"36603194\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Human ortholog lncRNA equivalence not established\", \"Binding region on EEF1D not mapped\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Identified the C-terminal GEF domain as essential for neurodevelopment, establishing a second, distinct disease mechanism from the alternatively spliced domain.\",\n      \"evidence\": \"Exome sequencing in two families with variant mapping to the GEF domain and clinical phenotyping\",\n      \"pmids\": [\"36576126\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Effect on GEF/eEF1A reactivation activity not functionally tested\", \"No animal or cellular model\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Defined post-transcriptional control of EEF1D, showing SRSF9 stabilizes its mRNA to drive cancer proliferation and metastasis.\",\n      \"evidence\": \"RNA immunoprecipitation, RNA pull-down, proteomics, and EEF1D knockdown rescue of SRSF9-driven phenotypes in colorectal cancer cells\",\n      \"pmids\": [\"38771720\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"3'UTR binding site not mapped\", \"Whether stabilization is isoform-selective unknown\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Defined post-translational control of EEF1D abundance through a competition between LGALS9B and the E3 ligase HERC5, linking stability directly to PI3K/AKT-driven tumor progression.\",\n      \"evidence\": \"Co-IP, competition binding assays, proteasome inhibitor experiments and in vitro/in vivo functional assays in gastric cancer\",\n      \"pmids\": [\"39639171\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Ubiquitination site on EEF1D not mapped\", \"Reciprocal validation in additional models lacking\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identified EEF1D as a proviral host factor that translocates to viral replication compartments during herpesvirus infection.\",\n      \"evidence\": \"Immunofluorescence co-localization with ICP8 and gD, siRNA knockdown, and viral titer measurement during BoAHV1 infection\",\n      \"pmids\": [\"40578269\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular function of EEF1D within replication compartments unknown\", \"Direct viral protein binding not demonstrated\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Showed that KSHV RTA suppresses EEF1D both by proteasomal degradation and promoter hypermethylation, establishing EEF1D as a host restriction factor for viral lytic reactivation.\",\n      \"evidence\": \"Co-IP, dual-luciferase reporters, DNMT3A-dependent methylation assays and EEF1D overexpression/knockdown with viral reactivation readouts\",\n      \"pmids\": [\"41805193\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which EEF1D restricts lytic replication not defined\", \"Single lab\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Established the molecular basis of EEF1B complex localization, showing exon-5-containing short isoforms anchor the complex to the ER via KTN1/RRBP1 independent of bulk translation.\",\n      \"evidence\": \"FLAG-EEF1D pulldown/MS, exon 5 deletion mutant, exon 5 KO mice, subcellular fractionation, multi-tissue abundance and global protein synthesis assays\",\n      \"pmids\": [\"42230146\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional purpose of ER anchoring (localized translation?) not established\", \"Connection between ER anchoring and disease/cancer phenotypes untested\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How EEF1D's distinct biochemical activities — GEF-mediated elongation, ER anchoring, and long-isoform heat-shock transcription — are coordinated, and which activity underlies its disease, metabolic, and cancer roles, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Isoform attribution of most cancer/metabolic phenotypes not defined\", \"No structural model linking GEF and transcriptional functions\", \"Direct demonstration of GEF activity loss in disease variants lacking\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [1, 7]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [8]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [13]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [13]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [7, 11]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [11, 13]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [8, 13]},\n      {\"term_id\": \"R-HSA-8953897\", \"supporting_discovery_ids\": [1, 7]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [3, 10]}\n    ],\n    \"complexes\": [\"eEF1B guanine nucleotide exchange complex\"],\n    \"partners\": [\"KTN1\", \"RRBP1\", \"LGALS9B\", \"HERC5\", \"SRSF9\", \"CK2\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"tie","faith_supported":7,"faith_total":8,"faith_pct":87.5}}