{"gene":"DHX30","run_date":"2026-06-09T23:54:42","timeline":{"discoveries":[{"year":2017,"finding":"De novo missense mutations in DHX30 located within conserved helicase motifs either impair ATPase activity or RNA recognition in vitro, and mutant protein variants exhibit an increased propensity to trigger stress granule (SG) formation, resulting in global translation inhibition.","method":"In vitro ATPase assays, RNA-binding assays, stress granule formation assays, global translation measurement","journal":"American Journal of Human Genetics","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — multiple orthogonal in vitro assays (ATPase, RNA recognition, SG formation, translation) on multiple independent patient-derived mutations in a single rigorous study","pmids":["29100085"],"is_preprint":false},{"year":2021,"finding":"DHX30 is established as an ATP-dependent RNA helicase; pathogenic missense variants within helicase core motifs impair both ATPase and helicase activity and trigger stress granule formation, interfering with global translation. Loss-of-function variants (haploinsufficiency/truncation) do not trigger SG formation and cause a milder phenotype, indicating that SG gain-of-function is the mechanism underlying severe disease. DHX30 is also an evolutionary conserved factor in SG assembly.","method":"ATPase assay, helicase activity assay, SG formation assay, global translation assay, CRISPR/Cas9 DHX30-deficient HEK293T and zebrafish models, in vivo behavioral assays","journal":"Genome Medicine","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — multiple orthogonal methods (ATPase, helicase, SG, translation) confirmed in both cell and zebrafish models, replicating and extending findings from PMID:29100085","pmids":["34020708"],"is_preprint":false},{"year":2010,"finding":"DHX30 interacts with the zinc-finger antiviral protein (ZAP) via their N-terminal domains, as demonstrated by pull-down and co-immunoprecipitation; shRNA-mediated knockdown of DHX30 reduces ZAP's antiviral activity, indicating DHX30 is required for optimal ZAP function in eliminating viral mRNAs.","method":"Pull-down assay, co-immunoprecipitation, shRNA knockdown with antiviral activity readout","journal":"Protein & Cell","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — reciprocal binding assays plus functional knockdown, single lab, two orthogonal methods","pmids":["21204022"],"is_preprint":false},{"year":2007,"finding":"Overexpression of DHX30 enhances HIV-1 gene expression but leads to generation of viruses that package significantly reduced levels of viral RNA and exhibit severely decreased infectivity, revealing an inhibitory role of DHX30 in HIV-1 RNA packaging.","method":"Overexpression in cell culture, viral RNA quantification, infectivity assay","journal":"Virology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — functional overexpression assay with multiple readouts (RNA packaging, infectivity), single lab","pmids":["18022663"],"is_preprint":false},{"year":2020,"finding":"DHX30, together with PCBP2, binds a 3' UTR CG-rich motif (CGPD-motif) present on mRNAs mediating p53-dependent apoptosis. In cells undergoing Nutlin-induced cell cycle arrest, the PCBP2-dependent binding of DHX30 to the CGPD-motif represses translation of these mRNAs. DHX30 depletion increases CGPD-motif mRNA translation and shifts the cellular response toward apoptosis; DHX30 inducible overexpression decreases translation of these mRNAs.","method":"Polysome profiling, RNA immunoprecipitation, shRNA knockdown and inducible overexpression with apoptosis/translation readouts","journal":"Cell Reports","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — polysome profiling plus RIP plus KD/OE functional assays, single lab, multiple orthogonal methods","pmids":["32234473"],"is_preprint":false},{"year":2021,"finding":"DHX30 exists as both a cytoplasmic and a more abundant mitochondrial isoform. Depletion of both isoforms in HCT116 cells constitutively alters polysome-associated mRNAs, enhancing translation of cytoplasmic ribosomal protein mRNAs while reducing translational efficiency of nuclear-encoded mitoribosome mRNAs, resulting in higher global translation, slower proliferation, and lower mitochondrial energy metabolism. Isoform-specific silencing supports a specific role for the cytoplasmic isoform in modulating global translation. RIP and eCLIP identified fourteen mitoribosome transcripts as direct DHX30 targets.","method":"Isoform-specific siRNA knockdown, polysome profiling, RIP, eCLIP, mitochondrial metabolic assays, proliferation assays in multiple cell lines","journal":"Cancers","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — eCLIP plus polysome profiling plus isoform-specific KD, multiple cell lines, single lab","pmids":["34503222"],"is_preprint":false},{"year":2022,"finding":"DHX30 is a component of mitochondrial RNA granules required for mitochondrial ribosome assembly and mitochondrial translation. ALS-linked mutant FUS interacts with DHX30, induces aberrant disulfide bond formation in DHX30 causing conformational change, promotes cytosolic mislocalization of DHX30 and its incorporation into stress granule-containing aggregates, and impairs mitochondrial translation and OXPHOS complex assembly — a phenotype similar to that caused by DHX30 knockdown alone.","method":"Co-immunoprecipitation, subcellular fractionation, detergent-solubility and density-gradient ultracentrifugation, blue-native gel electrophoresis, immunofluorescence, immunoelectron microscopy, DHX30 knockdown","journal":"Scientific Reports","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — multiple orthogonal methods (Co-IP, fractionation, BN-PAGE, EM) in single lab establishing DHX30 mitochondrial localization and its functional disruption by mutant FUS","pmids":["36163369"],"is_preprint":false},{"year":2022,"finding":"DHX30 is an intrinsic antiviral factor against Seneca Valley virus (SVV); it functions as a viral RNA-binding protein that inhibits SVV replication at the early stage of infection dependent on its helicase activity, inhibits double-stranded RNA production, and interacts with SVV 3D polymerase in an RNA-dependent manner. The SVV protease 3Cpro cleaves DHX30 at a specific site (dependent on 3Cpro protease activity) to antagonize its antiviral effects.","method":"LC-MS/MS, co-immunoprecipitation, RIP-seq, overexpression/knockdown with viral replication readouts, dsRNA quantification, protease cleavage assay","journal":"Journal of Virology","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — multiple methods (Co-IP, RIP-seq, functional KD/OE, cleavage assay), single lab","pmids":["36000840"],"is_preprint":false},{"year":2014,"finding":"HelG/DHX30 protein, purified from cells, exhibits helicase activity capable of unwinding DNA in vitro. Homozygous loss of helG/DHX30 in mice results in embryonic lethality by E9.5 with failure of somite differentiation and brain formation, demonstrating an essential role in early embryonic development.","method":"In vitro helicase/untwisting assay with purified protein, gene-trap homozygous mouse model with developmental phenotyping","journal":"Stem Cells and Development","confidence":"Medium","confidence_rationale":"Tier 1–3 / Moderate — in vitro helicase assay plus genetic mouse model, single lab, two orthogonal approaches","pmids":["25219788"],"is_preprint":false},{"year":2024,"finding":"DHX30 is recruited by lncRNA Anxa10-203 to form an Anxa10-203/DHX30 complex in the cytoplasm of trigeminal ganglion neurons, which enhances the stability of Mc1r mRNA, leading to upregulation of MC1R protein and increased intrinsic neuronal excitability.","method":"RNA pull-down, RNA immunoprecipitation, immunofluorescence, RNA-FISH, electrophysiology","journal":"The Journal of Headache and Pain","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, RNA pull-down and RIP without in vitro reconstitution or mutagenesis to confirm direct interaction mechanism","pmids":["38433184"],"is_preprint":false}],"current_model":"DHX30 is a DExH-box ATP-dependent RNA helicase with both cytoplasmic and mitochondrial isoforms: its cytoplasmic form represses translation of specific 3' UTR CGPD-motif mRNAs (in complex with PCBP2), modulates global ribosome biogenesis, and is required for optimal ZAP-mediated viral mRNA degradation and for restricting HIV-1 RNA packaging, while its mitochondrial form is a component of mitochondrial RNA granules essential for mitoribosome assembly and mitochondrial translation; pathogenic missense mutations in helicase core motifs abolish ATPase/helicase activity and confer a toxic gain-of-function by triggering stress granule assembly and global translation inhibition, causing a severe neurodevelopmental disorder."},"narrative":{"mechanistic_narrative":"DHX30 is a DExH-box ATP-dependent RNA helicase that governs translational output across both the cytoplasmic and mitochondrial compartments and functions as an antiviral effector [PMID:34020708, PMID:34503222]. In the cytoplasm, DHX30 acts with PCBP2 to bind a 3' UTR CG-rich motif (CGPD-motif) on p53-dependent apoptotic mRNAs and represses their translation, such that DHX30 depletion derepresses these transcripts and shifts cells toward apoptosis [PMID:32234473]; more broadly, the cytoplasmic isoform restrains global translation, and its loss enhances ribosomal protein mRNA translation while reducing translation of nuclear-encoded mitoribosome transcripts that it binds directly [PMID:34503222]. A distinct, more abundant mitochondrial isoform localizes to mitochondrial RNA granules where it is required for mitoribosome assembly, mitochondrial translation, and OXPHOS complex formation [PMID:36163369]. DHX30 additionally serves as an intrinsic antiviral factor: it is required for optimal ZAP-mediated viral mRNA elimination through an N-terminal interaction with ZAP [PMID:21204022], restricts HIV-1 RNA packaging [PMID:18022663], and inhibits Seneca Valley virus replication via helicase-dependent viral RNA binding and interaction with the viral 3D polymerase, an activity the viral 3Cpro protease cleaves to antagonize [PMID:36000840]. De novo missense mutations in conserved helicase core motifs abolish ATPase and helicase activity and confer a toxic gain-of-function that triggers stress granule assembly and global translation inhibition, causing a severe neurodevelopmental disorder, whereas loss-of-function variants produce a milder phenotype [PMID:29100085, PMID:34020708].","teleology":[{"year":2007,"claim":"Established the first functional role for DHX30, showing it inhibits HIV-1 RNA packaging despite enhancing viral gene expression, the earliest evidence linking the protein to RNA-virus biology.","evidence":"Overexpression in cell culture with viral RNA quantification and infectivity assays","pmids":["18022663"],"confidence":"Medium","gaps":["Mechanism by which DHX30 restricts packaging unresolved","No demonstration of direct viral RNA binding in this study","Reliance on overexpression rather than endogenous loss"]},{"year":2010,"claim":"Defined DHX30 as a cofactor of the zinc-finger antiviral protein ZAP, showing it is required for optimal ZAP-mediated elimination of viral mRNAs and extending its antiviral role to a defined protein partner.","evidence":"Pull-down and reciprocal co-immunoprecipitation mapping an N-terminal interaction, plus shRNA knockdown with antiviral readout","pmids":["21204022"],"confidence":"Medium","gaps":["Whether DHX30 helicase activity is needed for ZAP function not tested","Single lab","Structural basis of the N-terminal interaction unknown"]},{"year":2014,"claim":"Demonstrated catalytic helicase (unwinding) activity of purified DHX30 and its non-redundant requirement in early mammalian development.","evidence":"In vitro helicase/untwisting assay with purified protein and gene-trap homozygous mouse model","pmids":["25219788"],"confidence":"Medium","gaps":["Physiological RNA substrate not defined","Developmental lethality not mapped to a molecular function","DNA-unwinding readout leaves RNA preference open"]},{"year":2017,"claim":"Linked DHX30 to human disease, showing de novo missense mutations in helicase motifs impair ATPase or RNA recognition and promote stress granule formation with translation inhibition.","evidence":"In vitro ATPase and RNA-binding assays, stress granule and global translation assays on patient-derived variants","pmids":["29100085"],"confidence":"High","gaps":["Endogenous RNA targets in neurons not identified","SG composition and persistence not fully characterized","Genotype-phenotype correlation not yet established"]},{"year":2020,"claim":"Identified a sequence-specific translational repression mechanism, showing DHX30 with PCBP2 binds 3' UTR CGPD-motif mRNAs to repress p53-dependent apoptotic transcripts and tune the apoptosis-versus-arrest decision.","evidence":"Polysome profiling, RNA immunoprecipitation, shRNA knockdown and inducible overexpression with apoptosis/translation readouts","pmids":["32234473"],"confidence":"Medium","gaps":["Whether ATPase/helicase activity is required for repression not resolved","Direct vs PCBP2-bridged RNA contact not separated","In vivo relevance untested"]},{"year":2021,"claim":"Distinguished cytoplasmic and mitochondrial isoforms and established the cytoplasmic isoform as a modulator of global translation that directly binds nuclear-encoded mitoribosome transcripts.","evidence":"Isoform-specific siRNA knockdown, polysome profiling, RIP, eCLIP, and mitochondrial metabolic and proliferation assays in multiple cell lines","pmids":["34503222"],"confidence":"Medium","gaps":["Direct mechanism coupling translation control to proliferation not defined","Single lab","How the two isoforms are differentially generated/targeted unclear"]},{"year":2021,"claim":"Resolved the disease mechanism, showing gain-of-function SG-triggering missense variants cause severe disease while loss-of-function variants cause a milder phenotype, and confirmed DHX30 as a conserved SG factor in vivo.","evidence":"ATPase, helicase, SG and translation assays plus CRISPR DHX30-deficient HEK293T and zebrafish models with behavioral phenotyping","pmids":["34020708"],"confidence":"High","gaps":["Neuronal RNA targets driving pathology not defined","How SG assembly translates to developmental defects unknown","Therapeutic correction not addressed"]},{"year":2022,"claim":"Placed DHX30 in mitochondrial RNA granules as a requirement for mitoribosome assembly and translation, and showed ALS-mutant FUS mislocalizes DHX30 to stress-granule aggregates via aberrant disulfide-induced conformational change.","evidence":"Co-IP, subcellular fractionation, density-gradient and detergent-solubility analysis, blue-native PAGE, immunofluorescence, immunoelectron microscopy and DHX30 knockdown","pmids":["36163369"],"confidence":"Medium","gaps":["Identity of mitochondrial RNA substrates within granules not defined","Direct vs indirect role in mitoribosome assembly unresolved","Generality beyond mutant FUS context unknown"]},{"year":2022,"claim":"Showed DHX30 is an intrinsic antiviral RNA-binding factor against Seneca Valley virus acting through its helicase activity, and that the viral 3Cpro protease cleaves it to disable restriction.","evidence":"LC-MS/MS, co-IP, RIP-seq, overexpression/knockdown viral replication assays, dsRNA quantification and protease cleavage assay","pmids":["36000840"],"confidence":"Medium","gaps":["Whether antiviral function requires ZAP partnership not tested","Mechanism of dsRNA suppression not detailed","Single virus system"]},{"year":2024,"claim":"Implicated DHX30 in neuronal mRNA stabilization, acting as a partner recruited by lncRNA Anxa10-203 to stabilize Mc1r mRNA and increase neuronal excitability.","evidence":"RNA pull-down, RNA immunoprecipitation, immunofluorescence, RNA-FISH and electrophysiology in trigeminal ganglion neurons","pmids":["38433184"],"confidence":"Low","gaps":["RNA pull-down and RIP without reconstitution or mutagenesis to confirm direct interaction","Helicase-activity dependence not tested","Single lab, single physiological context"]},{"year":null,"claim":"How DHX30's distinct activities — sequence-specific translational repression, mitoribosome assembly, antiviral RNA surveillance and pathogenic SG nucleation — are coordinated by a shared catalytic mechanism and which endogenous RNA substrates drive each remains unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["No unifying structural model of substrate engagement","Comprehensive endogenous RNA target map across compartments lacking","Link between molecular activity and the neurodevelopmental phenotype not mechanistically closed"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140098","term_label":"catalytic activity, acting on RNA","supporting_discovery_ids":[0,1,8]},{"term_id":"GO:0140657","term_label":"ATP-dependent activity","supporting_discovery_ids":[0,1]},{"term_id":"GO:0003723","term_label":"RNA binding","supporting_discovery_ids":[4,5,7]},{"term_id":"GO:0045182","term_label":"translation regulator activity","supporting_discovery_ids":[4,5]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[4,5,9]},{"term_id":"GO:0005739","term_label":"mitochondrion","supporting_discovery_ids":[5,6]}],"pathway":[{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[2,3,7]},{"term_id":"R-HSA-8953854","term_label":"Metabolism of RNA","supporting_discovery_ids":[5,6]}],"complexes":["mitochondrial RNA granule","DHX30-PCBP2 CGPD-motif repressor complex","Anxa10-203/DHX30 lncRNA complex"],"partners":["PCBP2","ZAP","FUS"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q7L2E3","full_name":"ATP-dependent RNA helicase DHX30","aliases":["DEAH box protein 30"],"length_aa":1194,"mass_kda":133.9,"function":"RNA-dependent helicase (PubMed:29100085). Plays an important role in the assembly of the mitochondrial large ribosomal subunit (PubMed:25683715, PubMed:29100085). Required for optimal function of the zinc-finger antiviral protein ZC3HAV1 (By similarity). Associates with mitochondrial DNA (PubMed:18063578). Involved in nervous system development and differentiation through its involvement in the up-regulation of a number of genes which are required for neurogenesis, including GSC, NCAM1, neurogenin, and NEUROD (By similarity)","subcellular_location":"Cytoplasm; Mitochondrion; Mitochondrion matrix, mitochondrion nucleoid","url":"https://www.uniprot.org/uniprotkb/Q7L2E3/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/DHX30","classification":"Not Classified","n_dependent_lines":256,"n_total_lines":1208,"dependency_fraction":0.2119205298013245},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"CALM3","stoichiometry":0.2},{"gene":"CAPRIN1","stoichiometry":0.2},{"gene":"CAPZB","stoichiometry":0.2},{"gene":"CTCF","stoichiometry":0.2},{"gene":"DDX21","stoichiometry":0.2},{"gene":"DHX9","stoichiometry":0.2},{"gene":"DRG1","stoichiometry":0.2},{"gene":"G3BP2","stoichiometry":0.2},{"gene":"GSPT1","stoichiometry":0.2},{"gene":"HMGB2","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/DHX30","total_profiled":1310},"omim":[{"mim_id":"617804","title":"NEURODEVELOPMENTAL DISORDER WITH VARIABLE MOTOR AND LANGUAGE IMPAIRMENT; NEDMIAL","url":"https://www.omim.org/entry/617804"},{"mim_id":"616423","title":"DExH-BOX HELICASE 30; DHX30","url":"https://www.omim.org/entry/616423"},{"mim_id":"616422","title":"TRANSCRIPTION ELONGATION FACTOR, MITOCHONDRIAL; TEFM","url":"https://www.omim.org/entry/616422"},{"mim_id":"614918","title":"PENTATRICOPEPTIDE REPEAT DOMAIN-CONTAINING PROTEIN 3; PTCD3","url":"https://www.omim.org/entry/614918"},{"mim_id":"607312","title":"ZINC FINGER CCCH DOMAIN-CONTAINING ANTIVIRAL PROTEIN 1; ZC3HAV1","url":"https://www.omim.org/entry/607312"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Mitochondria","reliability":"Supported"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/DHX30"},"hgnc":{"alias_symbol":["KIAA0890","FLJ11214"],"prev_symbol":["DDX30"]},"alphafold":{"accession":"Q7L2E3","domains":[{"cath_id":"3.30.160.20","chopping":"47-146","consensus_level":"high","plddt":85.9962,"start":47,"end":146},{"cath_id":"3.30.160.20","chopping":"233-326","consensus_level":"high","plddt":86.9577,"start":233,"end":326},{"cath_id":"3.40.50.300","chopping":"452-607","consensus_level":"high","plddt":91.7392,"start":452,"end":607},{"cath_id":"3.40.50.300","chopping":"619-635_652-808","consensus_level":"high","plddt":87.6221,"start":619,"end":808},{"cath_id":"-","chopping":"855-1009","consensus_level":"medium","plddt":90.4767,"start":855,"end":1009},{"cath_id":"-","chopping":"1024-1181","consensus_level":"medium","plddt":87.448,"start":1024,"end":1181},{"cath_id":"1.10.10","chopping":"817-853","consensus_level":"high","plddt":92.9941,"start":817,"end":853}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q7L2E3","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q7L2E3-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q7L2E3-F1-predicted_aligned_error_v6.png","plddt_mean":81.06},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=DHX30","jax_strain_url":"https://www.jax.org/strain/search?query=DHX30"},"sequence":{"accession":"Q7L2E3","fasta_url":"https://rest.uniprot.org/uniprotkb/Q7L2E3.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q7L2E3/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q7L2E3"}},"corpus_meta":[{"pmid":"29100085","id":"PMC_29100085","title":"De Novo Missense Mutations in DHX30 Impair Global Translation and Cause a Neurodevelopmental Disorder.","date":"2017","source":"American journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/29100085","citation_count":71,"is_preprint":false},{"pmid":"21204022","id":"PMC_21204022","title":"DEXH-Box protein DHX30 is required for optimal function of the zinc-finger antiviral protein.","date":"2010","source":"Protein & cell","url":"https://pubmed.ncbi.nlm.nih.gov/21204022","citation_count":43,"is_preprint":false},{"pmid":"18022663","id":"PMC_18022663","title":"The packaging of human immunodeficiency virus type 1 RNA is restricted by overexpression of an RNA helicase DHX30.","date":"2007","source":"Virology","url":"https://pubmed.ncbi.nlm.nih.gov/18022663","citation_count":27,"is_preprint":false},{"pmid":"34020708","id":"PMC_34020708","title":"Genotype-phenotype correlations and novel molecular insights into the DHX30-associated neurodevelopmental disorders.","date":"2021","source":"Genome medicine","url":"https://pubmed.ncbi.nlm.nih.gov/34020708","citation_count":24,"is_preprint":false},{"pmid":"32234473","id":"PMC_32234473","title":"Nutlin-Induced Apoptosis Is Specified by a Translation Program Regulated by PCBP2 and DHX30.","date":"2020","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/32234473","citation_count":24,"is_preprint":false},{"pmid":"34503222","id":"PMC_34503222","title":"DHX30 Coordinates Cytoplasmic Translation and Mitochondrial Function Contributing to Cancer Cell Survival.","date":"2021","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/34503222","citation_count":17,"is_preprint":false},{"pmid":"25219788","id":"PMC_25219788","title":"The novel helicase helG (DHX30) is expressed during gastrulation in mice and has a structure similar to a human DExH box helicase.","date":"2014","source":"Stem cells and development","url":"https://pubmed.ncbi.nlm.nih.gov/25219788","citation_count":16,"is_preprint":false},{"pmid":"36000840","id":"PMC_36000840","title":"Seneca Valley Virus Induces DHX30 Cleavage to Antagonize Its Antiviral Effects.","date":"2022","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/36000840","citation_count":13,"is_preprint":false},{"pmid":"36163369","id":"PMC_36163369","title":"Conformational change of RNA-helicase DHX30 by ALS/FTD-linked FUS induces mitochondrial dysfunction and cytosolic aggregates.","date":"2022","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/36163369","citation_count":9,"is_preprint":false},{"pmid":"38433184","id":"PMC_38433184","title":"LncRNA Anxa10-203 enhances Mc1r mRNA stability to promote neuropathic pain by recruiting DHX30 in the trigeminal ganglion.","date":"2024","source":"The journal of headache and pain","url":"https://pubmed.ncbi.nlm.nih.gov/38433184","citation_count":7,"is_preprint":false},{"pmid":"36643085","id":"PMC_36643085","title":"A Novel De Novo Mutation of the DHX30 Gene in a Patient With Neurodevelopmental Disorder, Severe Motor Impairment, and Absent Language (NEDMIAL).","date":"2023","source":"Cureus","url":"https://pubmed.ncbi.nlm.nih.gov/36643085","citation_count":3,"is_preprint":false},{"pmid":"38366977","id":"PMC_38366977","title":"A de novo pathogenic variant in DHX30 gene in a fetus with isolated dysgenesis of the corpus callosum.","date":"2024","source":"Prenatal diagnosis","url":"https://pubmed.ncbi.nlm.nih.gov/38366977","citation_count":1,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":7647,"output_tokens":2755,"usd":0.032133,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":10065,"output_tokens":3810,"usd":0.072788,"stage2_stop_reason":"end_turn"},"total_usd":0.104921,"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\": 2017,\n      \"finding\": \"De novo missense mutations in DHX30 located within conserved helicase motifs either impair ATPase activity or RNA recognition in vitro, and mutant protein variants exhibit an increased propensity to trigger stress granule (SG) formation, resulting in global translation inhibition.\",\n      \"method\": \"In vitro ATPase assays, RNA-binding assays, stress granule formation assays, global translation measurement\",\n      \"journal\": \"American Journal of Human Genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — multiple orthogonal in vitro assays (ATPase, RNA recognition, SG formation, translation) on multiple independent patient-derived mutations in a single rigorous study\",\n      \"pmids\": [\"29100085\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"DHX30 is established as an ATP-dependent RNA helicase; pathogenic missense variants within helicase core motifs impair both ATPase and helicase activity and trigger stress granule formation, interfering with global translation. Loss-of-function variants (haploinsufficiency/truncation) do not trigger SG formation and cause a milder phenotype, indicating that SG gain-of-function is the mechanism underlying severe disease. DHX30 is also an evolutionary conserved factor in SG assembly.\",\n      \"method\": \"ATPase assay, helicase activity assay, SG formation assay, global translation assay, CRISPR/Cas9 DHX30-deficient HEK293T and zebrafish models, in vivo behavioral assays\",\n      \"journal\": \"Genome Medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — multiple orthogonal methods (ATPase, helicase, SG, translation) confirmed in both cell and zebrafish models, replicating and extending findings from PMID:29100085\",\n      \"pmids\": [\"34020708\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"DHX30 interacts with the zinc-finger antiviral protein (ZAP) via their N-terminal domains, as demonstrated by pull-down and co-immunoprecipitation; shRNA-mediated knockdown of DHX30 reduces ZAP's antiviral activity, indicating DHX30 is required for optimal ZAP function in eliminating viral mRNAs.\",\n      \"method\": \"Pull-down assay, co-immunoprecipitation, shRNA knockdown with antiviral activity readout\",\n      \"journal\": \"Protein & Cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — reciprocal binding assays plus functional knockdown, single lab, two orthogonal methods\",\n      \"pmids\": [\"21204022\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Overexpression of DHX30 enhances HIV-1 gene expression but leads to generation of viruses that package significantly reduced levels of viral RNA and exhibit severely decreased infectivity, revealing an inhibitory role of DHX30 in HIV-1 RNA packaging.\",\n      \"method\": \"Overexpression in cell culture, viral RNA quantification, infectivity assay\",\n      \"journal\": \"Virology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — functional overexpression assay with multiple readouts (RNA packaging, infectivity), single lab\",\n      \"pmids\": [\"18022663\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"DHX30, together with PCBP2, binds a 3' UTR CG-rich motif (CGPD-motif) present on mRNAs mediating p53-dependent apoptosis. In cells undergoing Nutlin-induced cell cycle arrest, the PCBP2-dependent binding of DHX30 to the CGPD-motif represses translation of these mRNAs. DHX30 depletion increases CGPD-motif mRNA translation and shifts the cellular response toward apoptosis; DHX30 inducible overexpression decreases translation of these mRNAs.\",\n      \"method\": \"Polysome profiling, RNA immunoprecipitation, shRNA knockdown and inducible overexpression with apoptosis/translation readouts\",\n      \"journal\": \"Cell Reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — polysome profiling plus RIP plus KD/OE functional assays, single lab, multiple orthogonal methods\",\n      \"pmids\": [\"32234473\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"DHX30 exists as both a cytoplasmic and a more abundant mitochondrial isoform. Depletion of both isoforms in HCT116 cells constitutively alters polysome-associated mRNAs, enhancing translation of cytoplasmic ribosomal protein mRNAs while reducing translational efficiency of nuclear-encoded mitoribosome mRNAs, resulting in higher global translation, slower proliferation, and lower mitochondrial energy metabolism. Isoform-specific silencing supports a specific role for the cytoplasmic isoform in modulating global translation. RIP and eCLIP identified fourteen mitoribosome transcripts as direct DHX30 targets.\",\n      \"method\": \"Isoform-specific siRNA knockdown, polysome profiling, RIP, eCLIP, mitochondrial metabolic assays, proliferation assays in multiple cell lines\",\n      \"journal\": \"Cancers\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — eCLIP plus polysome profiling plus isoform-specific KD, multiple cell lines, single lab\",\n      \"pmids\": [\"34503222\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"DHX30 is a component of mitochondrial RNA granules required for mitochondrial ribosome assembly and mitochondrial translation. ALS-linked mutant FUS interacts with DHX30, induces aberrant disulfide bond formation in DHX30 causing conformational change, promotes cytosolic mislocalization of DHX30 and its incorporation into stress granule-containing aggregates, and impairs mitochondrial translation and OXPHOS complex assembly — a phenotype similar to that caused by DHX30 knockdown alone.\",\n      \"method\": \"Co-immunoprecipitation, subcellular fractionation, detergent-solubility and density-gradient ultracentrifugation, blue-native gel electrophoresis, immunofluorescence, immunoelectron microscopy, DHX30 knockdown\",\n      \"journal\": \"Scientific Reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — multiple orthogonal methods (Co-IP, fractionation, BN-PAGE, EM) in single lab establishing DHX30 mitochondrial localization and its functional disruption by mutant FUS\",\n      \"pmids\": [\"36163369\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"DHX30 is an intrinsic antiviral factor against Seneca Valley virus (SVV); it functions as a viral RNA-binding protein that inhibits SVV replication at the early stage of infection dependent on its helicase activity, inhibits double-stranded RNA production, and interacts with SVV 3D polymerase in an RNA-dependent manner. The SVV protease 3Cpro cleaves DHX30 at a specific site (dependent on 3Cpro protease activity) to antagonize its antiviral effects.\",\n      \"method\": \"LC-MS/MS, co-immunoprecipitation, RIP-seq, overexpression/knockdown with viral replication readouts, dsRNA quantification, protease cleavage assay\",\n      \"journal\": \"Journal of Virology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — multiple methods (Co-IP, RIP-seq, functional KD/OE, cleavage assay), single lab\",\n      \"pmids\": [\"36000840\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"HelG/DHX30 protein, purified from cells, exhibits helicase activity capable of unwinding DNA in vitro. Homozygous loss of helG/DHX30 in mice results in embryonic lethality by E9.5 with failure of somite differentiation and brain formation, demonstrating an essential role in early embryonic development.\",\n      \"method\": \"In vitro helicase/untwisting assay with purified protein, gene-trap homozygous mouse model with developmental phenotyping\",\n      \"journal\": \"Stem Cells and Development\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1–3 / Moderate — in vitro helicase assay plus genetic mouse model, single lab, two orthogonal approaches\",\n      \"pmids\": [\"25219788\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"DHX30 is recruited by lncRNA Anxa10-203 to form an Anxa10-203/DHX30 complex in the cytoplasm of trigeminal ganglion neurons, which enhances the stability of Mc1r mRNA, leading to upregulation of MC1R protein and increased intrinsic neuronal excitability.\",\n      \"method\": \"RNA pull-down, RNA immunoprecipitation, immunofluorescence, RNA-FISH, electrophysiology\",\n      \"journal\": \"The Journal of Headache and Pain\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, RNA pull-down and RIP without in vitro reconstitution or mutagenesis to confirm direct interaction mechanism\",\n      \"pmids\": [\"38433184\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"DHX30 is a DExH-box ATP-dependent RNA helicase with both cytoplasmic and mitochondrial isoforms: its cytoplasmic form represses translation of specific 3' UTR CGPD-motif mRNAs (in complex with PCBP2), modulates global ribosome biogenesis, and is required for optimal ZAP-mediated viral mRNA degradation and for restricting HIV-1 RNA packaging, while its mitochondrial form is a component of mitochondrial RNA granules essential for mitoribosome assembly and mitochondrial translation; pathogenic missense mutations in helicase core motifs abolish ATPase/helicase activity and confer a toxic gain-of-function by triggering stress granule assembly and global translation inhibition, causing a severe neurodevelopmental disorder.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"DHX30 is a DExH-box ATP-dependent RNA helicase that governs translational output across both the cytoplasmic and mitochondrial compartments and functions as an antiviral effector [#1, #5]. In the cytoplasm, DHX30 acts with PCBP2 to bind a 3' UTR CG-rich motif (CGPD-motif) on p53-dependent apoptotic mRNAs and represses their translation, such that DHX30 depletion derepresses these transcripts and shifts cells toward apoptosis [#4]; more broadly, the cytoplasmic isoform restrains global translation, and its loss enhances ribosomal protein mRNA translation while reducing translation of nuclear-encoded mitoribosome transcripts that it binds directly [#5]. A distinct, more abundant mitochondrial isoform localizes to mitochondrial RNA granules where it is required for mitoribosome assembly, mitochondrial translation, and OXPHOS complex formation [#6]. DHX30 additionally serves as an intrinsic antiviral factor: it is required for optimal ZAP-mediated viral mRNA elimination through an N-terminal interaction with ZAP [#2], restricts HIV-1 RNA packaging [#3], and inhibits Seneca Valley virus replication via helicase-dependent viral RNA binding and interaction with the viral 3D polymerase, an activity the viral 3Cpro protease cleaves to antagonize [#7]. De novo missense mutations in conserved helicase core motifs abolish ATPase and helicase activity and confer a toxic gain-of-function that triggers stress granule assembly and global translation inhibition, causing a severe neurodevelopmental disorder, whereas loss-of-function variants produce a milder phenotype [#0, #1].\",\n  \"teleology\": [\n    {\n      \"year\": 2007,\n      \"claim\": \"Established the first functional role for DHX30, showing it inhibits HIV-1 RNA packaging despite enhancing viral gene expression, the earliest evidence linking the protein to RNA-virus biology.\",\n      \"evidence\": \"Overexpression in cell culture with viral RNA quantification and infectivity assays\",\n      \"pmids\": [\"18022663\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which DHX30 restricts packaging unresolved\", \"No demonstration of direct viral RNA binding in this study\", \"Reliance on overexpression rather than endogenous loss\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Defined DHX30 as a cofactor of the zinc-finger antiviral protein ZAP, showing it is required for optimal ZAP-mediated elimination of viral mRNAs and extending its antiviral role to a defined protein partner.\",\n      \"evidence\": \"Pull-down and reciprocal co-immunoprecipitation mapping an N-terminal interaction, plus shRNA knockdown with antiviral readout\",\n      \"pmids\": [\"21204022\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether DHX30 helicase activity is needed for ZAP function not tested\", \"Single lab\", \"Structural basis of the N-terminal interaction unknown\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Demonstrated catalytic helicase (unwinding) activity of purified DHX30 and its non-redundant requirement in early mammalian development.\",\n      \"evidence\": \"In vitro helicase/untwisting assay with purified protein and gene-trap homozygous mouse model\",\n      \"pmids\": [\"25219788\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Physiological RNA substrate not defined\", \"Developmental lethality not mapped to a molecular function\", \"DNA-unwinding readout leaves RNA preference open\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Linked DHX30 to human disease, showing de novo missense mutations in helicase motifs impair ATPase or RNA recognition and promote stress granule formation with translation inhibition.\",\n      \"evidence\": \"In vitro ATPase and RNA-binding assays, stress granule and global translation assays on patient-derived variants\",\n      \"pmids\": [\"29100085\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Endogenous RNA targets in neurons not identified\", \"SG composition and persistence not fully characterized\", \"Genotype-phenotype correlation not yet established\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Identified a sequence-specific translational repression mechanism, showing DHX30 with PCBP2 binds 3' UTR CGPD-motif mRNAs to repress p53-dependent apoptotic transcripts and tune the apoptosis-versus-arrest decision.\",\n      \"evidence\": \"Polysome profiling, RNA immunoprecipitation, shRNA knockdown and inducible overexpression with apoptosis/translation readouts\",\n      \"pmids\": [\"32234473\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether ATPase/helicase activity is required for repression not resolved\", \"Direct vs PCBP2-bridged RNA contact not separated\", \"In vivo relevance untested\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Distinguished cytoplasmic and mitochondrial isoforms and established the cytoplasmic isoform as a modulator of global translation that directly binds nuclear-encoded mitoribosome transcripts.\",\n      \"evidence\": \"Isoform-specific siRNA knockdown, polysome profiling, RIP, eCLIP, and mitochondrial metabolic and proliferation assays in multiple cell lines\",\n      \"pmids\": [\"34503222\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct mechanism coupling translation control to proliferation not defined\", \"Single lab\", \"How the two isoforms are differentially generated/targeted unclear\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Resolved the disease mechanism, showing gain-of-function SG-triggering missense variants cause severe disease while loss-of-function variants cause a milder phenotype, and confirmed DHX30 as a conserved SG factor in vivo.\",\n      \"evidence\": \"ATPase, helicase, SG and translation assays plus CRISPR DHX30-deficient HEK293T and zebrafish models with behavioral phenotyping\",\n      \"pmids\": [\"34020708\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Neuronal RNA targets driving pathology not defined\", \"How SG assembly translates to developmental defects unknown\", \"Therapeutic correction not addressed\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Placed DHX30 in mitochondrial RNA granules as a requirement for mitoribosome assembly and translation, and showed ALS-mutant FUS mislocalizes DHX30 to stress-granule aggregates via aberrant disulfide-induced conformational change.\",\n      \"evidence\": \"Co-IP, subcellular fractionation, density-gradient and detergent-solubility analysis, blue-native PAGE, immunofluorescence, immunoelectron microscopy and DHX30 knockdown\",\n      \"pmids\": [\"36163369\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Identity of mitochondrial RNA substrates within granules not defined\", \"Direct vs indirect role in mitoribosome assembly unresolved\", \"Generality beyond mutant FUS context unknown\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Showed DHX30 is an intrinsic antiviral RNA-binding factor against Seneca Valley virus acting through its helicase activity, and that the viral 3Cpro protease cleaves it to disable restriction.\",\n      \"evidence\": \"LC-MS/MS, co-IP, RIP-seq, overexpression/knockdown viral replication assays, dsRNA quantification and protease cleavage assay\",\n      \"pmids\": [\"36000840\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether antiviral function requires ZAP partnership not tested\", \"Mechanism of dsRNA suppression not detailed\", \"Single virus system\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Implicated DHX30 in neuronal mRNA stabilization, acting as a partner recruited by lncRNA Anxa10-203 to stabilize Mc1r mRNA and increase neuronal excitability.\",\n      \"evidence\": \"RNA pull-down, RNA immunoprecipitation, immunofluorescence, RNA-FISH and electrophysiology in trigeminal ganglion neurons\",\n      \"pmids\": [\"38433184\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"RNA pull-down and RIP without reconstitution or mutagenesis to confirm direct interaction\", \"Helicase-activity dependence not tested\", \"Single lab, single physiological context\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How DHX30's distinct activities — sequence-specific translational repression, mitoribosome assembly, antiviral RNA surveillance and pathogenic SG nucleation — are coordinated by a shared catalytic mechanism and which endogenous RNA substrates drive each remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No unifying structural model of substrate engagement\", \"Comprehensive endogenous RNA target map across compartments lacking\", \"Link between molecular activity and the neurodevelopmental phenotype not mechanistically closed\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140098\", \"supporting_discovery_ids\": [0, 1, 8]},\n      {\"term_id\": \"GO:0140657\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"GO:0003723\", \"supporting_discovery_ids\": [4, 5, 7]},\n      {\"term_id\": \"GO:0045182\", \"supporting_discovery_ids\": [4, 5]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [4, 5, 9]},\n      {\"term_id\": \"GO:0005739\", \"supporting_discovery_ids\": [5, 6]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-72766\", \"supporting_discovery_ids\": [4, 5]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [2, 3, 7]},\n      {\"term_id\": \"R-HSA-8953854\", \"supporting_discovery_ids\": [5, 6]}\n    ],\n    \"complexes\": [\n      \"mitochondrial RNA granule\",\n      \"DHX30-PCBP2 CGPD-motif repressor complex\",\n      \"Anxa10-203/DHX30 lncRNA complex\"\n    ],\n    \"partners\": [\n      \"PCBP2\",\n      \"ZAP\",\n      \"FUS\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":5,"faith_pct":100.0}}