{"gene":"DHX33","run_date":"2026-04-28T17:46:02","timeline":{"discoveries":[{"year":2011,"finding":"DHX33 is a cell cycle-regulated nucleolar protein that associates with ribosomal DNA (rDNA) loci and interacts with the RNA Pol I transcription factor UBF; DHX33 knockdown decreases Pol I association with rDNA and dramatically reduces 47S rRNA synthesis, while an NTPase-defective mutant (K94R) acts as a dominant negative inhibitor of rRNA synthesis.","method":"lentiviral RNAi screen, ChIP, Co-IP, mutagenesis (K94R dominant negative, DNA-binding defective mutant), rRNA synthesis assays","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 1–2 — multiple orthogonal methods including mutagenesis, ChIP, Co-IP, and functional assays in a single study","pmids":["21930779"],"is_preprint":false},{"year":2013,"finding":"DHX33 acts as a cytosolic RNA sensor that binds dsRNA via its helicase C domain, interacts with NLRP3, and forms the inflammasome complex following RNA stimulation, leading to caspase-1 activation and IL-1β/IL-18 secretion in human macrophages.","method":"shRNA knockdown, binding domain mapping, Co-IP (DHX33–NLRP3 interaction), caspase-1 activity assay, cytokine secretion assay","journal":"Immunity","confidence":"High","confidence_rationale":"Tier 2 — domain-mapping pulldown, Co-IP, and functional inflammasome assays; replicated in independent study (PMID:24037184)","pmids":["23871209"],"is_preprint":false},{"year":2013,"finding":"DHX33 interacts with IPS-1/MAVS in myeloid dendritic cells via the HELICc domain of DHX33 and the C-terminal domain of IPS-1, independently of RIG-I/MDA5, to mediate dsRNA-induced type I IFN production and activation of MAP kinases, NF-κB, and IRF3.","method":"shRNA knockdown, domain-mapping Co-IP (HELICc–IPS-1 C-terminal), poly I:C binding assay, signaling pathway analysis","journal":"Cellular & molecular immunology","confidence":"High","confidence_rationale":"Tier 2 — reciprocal domain-mapping Co-IP with functional signaling readouts; consistent with PMID:23871209","pmids":["24037184"],"is_preprint":false},{"year":2013,"finding":"DHX33 protein translation is regulated oppositely by ARF (which reduces polysome-associated DHX33 mRNA) and RasV12 (which shifts DHX33 mRNA to actively translating polysomes via PI3K/mTOR/MAPK pathways); DHX33 is required downstream of RasV12 for enhanced rRNA transcription and cellular transformation.","method":"polysome fractionation, translational reporter assays, kinase inhibitor treatment, DHX33 knockdown rescue experiments, in vitro and in vivo transformation assays","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods including polysome profiling, pharmacological inhibition, and in vivo tumor models","pmids":["23401854"],"is_preprint":false},{"year":2015,"finding":"DHX33 promotes mRNA translation initiation by facilitating elongation-competent 80S ribosome assembly; DHX33 reduction markedly reduces polyribosome formation and causes global inhibition of mRNA translation, with an accumulation of mRNAs stalled at the 80S ribosome; helicase-defective DHX33 cannot rescue this phenotype.","method":"polyribosome profiling, RNA immunoprecipitation (RIP), ribosomal protein/translation factor Co-IP, helicase-dead mutant rescue","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods including polysome profiling, RIP, and mutagenesis with rescue","pmids":["26100019"],"is_preprint":false},{"year":2016,"finding":"DHX33 physically associates with promoters of cell cycle genes (cyclins, E2F1, CDC, MCM genes) and controls loading of active RNA polymerase II onto these promoters to drive cell cycle progression; CRISPR-mediated knockout in zebrafish confirmed downregulation of these targets in vivo.","method":"ChIP, RNA Pol II ChIP, siRNA knockdown, cell cycle analysis, CRISPR/Cas9 knockout in zebrafish, xenograft model","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 — ChIP-based mechanism with multiple orthogonal validations including in vivo zebrafish model","pmids":["27601587"],"is_preprint":false},{"year":2017,"finding":"c-Myc binds to the DHX33 upstream promoter region and stimulates DHX33 transcription; DHX33 in turn promotes transcription of MMP9, MMP14, and PLAU by directly binding to their promoters, thereby promoting cancer cell migration.","method":"ChIP (Myc at DHX33 promoter; DHX33 at MMP/PLAU promoters), luciferase reporter, knockdown/rescue, in vivo mouse leukemia model","journal":"Carcinogenesis","confidence":"Medium","confidence_rationale":"Tier 2 — ChIP and functional assays from a single lab","pmids":["28498893"],"is_preprint":false},{"year":2017,"finding":"USP36 deubiquitinase reduces ubiquitination of DHX33 and increases its protein stability; loss of USP36 destabilizes DHX33, impairs rRNA synthesis and protein translation, and causes preimplantation lethality in mice.","method":"ubiquitination assay, protein stability assay, USP36 knockout mouse, shRNA knockdown, Northern blot, O-propargyl-puromycin incorporation, electron microscopy","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 — in vivo genetic model combined with biochemical ubiquitination assays and multiple orthogonal methods","pmids":["29273634"],"is_preprint":false},{"year":2018,"finding":"Purified recombinant DHX33 protein possesses ATPase activity stimulated by DNA or RNA duplexes, and this ATPase activity is coupled to unwinding of both RNA and DNA duplexes; mutation of a key residue in the ATP-binding site abolishes unwinding activity.","method":"in vitro ATPase assay with purified recombinant protein, helicase unwinding assay, ATP-binding site mutagenesis","journal":"Biochemistry","confidence":"High","confidence_rationale":"Tier 1 — in vitro biochemical reconstitution with mutagenesis of active site","pmids":["29870660"],"is_preprint":false},{"year":2018,"finding":"DHX33 is required for glioblastoma cell proliferation and migration; overexpression of wild-type DHX33 but not a helicase-dead mutant confers resistance to mTOR inhibitors, indicating the helicase activity is required for this function.","method":"DHX33 knockdown (proliferation/migration assays, xenograft), wild-type vs. helicase-dead mutant rescue, mTOR inhibitor treatment","journal":"Cellular signalling","confidence":"Medium","confidence_rationale":"Tier 2 — mutagenesis with functional rescue in cancer cell context, single lab","pmids":["30552990"],"is_preprint":false},{"year":2018,"finding":"DHX33 produces two protein isoforms of different size from two in-frame start codons via alternative translation initiation (leaky scanning); both isoforms have similar cellular localization and functions.","method":"mutagenesis of start codons, cell line and mouse model analysis, immunoblotting","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 — genetic mutagenesis with multiple model validations, single lab","pmids":["29864424"],"is_preprint":false},{"year":2019,"finding":"A short DHX33 variant (DHX33-2, 534 aa comprising the C-terminal helicase domain) localizes preferentially to the cytoplasm and interacts with DDX3, eIF3, hnRNPs, and poly(A)-binding protein to stimulate translation of a subset of mRNAs involved in cell proliferation.","method":"protein immunoprecipitation, RIP-seq, RNA sequencing, subcellular fractionation","journal":"Journal of cellular physiology","confidence":"Medium","confidence_rationale":"Tier 2–3 — Co-IP and RIP with functional translation assays, single lab","pmids":["30684270"],"is_preprint":false},{"year":2020,"finding":"DHX33 recruits Gadd45a and DNA dioxygenase Tet1 to promoters of specific genes (including glycolytic genes LDHA, PDK1, PKM2), causing local DNA demethylation (reduced 5-hydroxymethylcytosine) and enhanced histone H4 acetylation to promote their transcription; this involves DHX33 binding to CG-rich promoter regions, potentially via R-loop formation.","method":"ChIP, 5hmC quantification, Co-IP (DHX33–AP-2β–Gadd45a complex), RNA-seq, promoter methylation analysis, DHX33 knockdown","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods (ChIP, 5hmC assay, Co-IP, RNA-seq) establishing a mechanistic epigenetic pathway","pmids":["32312884","32617965"],"is_preprint":false},{"year":2022,"finding":"GSK-3β directly phosphorylates DHX33 at T482, triggering ubiquitination-mediated protein degradation; a major ubiquitination site was identified at K94 (also critical for ATP binding/helicase activity); cancer cells with frequent GSK-3β inactivation have elevated DHX33 stability.","method":"in vitro kinase assay, phosphorylation site mutagenesis (T482), ubiquitination assay, protein stability assay, GSK-3β inhibitor/activator treatment","journal":"Cellular signalling","confidence":"High","confidence_rationale":"Tier 1–2 — in vitro kinase assay with mutagenesis and ubiquitination assay, single lab but multiple orthogonal methods","pmids":["36403931"],"is_preprint":false},{"year":2023,"finding":"Dhx33 is required for activation-induced upregulation of ribosomal DNA transcription in B cells; B-cell-specific deletion of Dhx33 impairs B-cell growth and proliferation after activation, causing nucleolar stress, p53 accumulation, and cellular death, without affecting steady-state B-cell function.","method":"CRISPR/Cas9-mediated B-cell-specific knockout mouse, rDNA transcription assay, 47S rRNA measurement, p53 immunoblot, germinal center analysis","journal":"Cellular & molecular immunology","confidence":"High","confidence_rationale":"Tier 2 — clean conditional knockout in vivo with mechanistic rRNA and p53 pathway readouts","pmids":["36631557"],"is_preprint":false},{"year":2023,"finding":"DHX33 promotes expression of mevalonate pathway genes downstream of mutant p53 and Ras; in vivo lung tumors carrying mutant p53/KrasG12D show upregulated mevalonate pathway genes that are debilitated upon DHX33 loss.","method":"DHX33 knockdown, in vivo KrasG12D/p53-mutant mouse model, gene expression analysis","journal":"Biochimica et biophysica acta. General subjects","confidence":"Medium","confidence_rationale":"Tier 2 — in vivo genetic epistasis with pathway gene readout, single lab","pmids":["38143011"],"is_preprint":false},{"year":2024,"finding":"DHX33 promotes expression of lipid metabolism genes FADS1, FADS2, and SCD1, sensitizing cancer cells to ferroptosis; pharmacological inhibition of DHX33 by KY386 induces ferroptosis-mediated cancer cell death.","method":"DHX33 inhibitor (KY386) treatment, ferroptosis pathway assays, gene expression analysis, cell viability assays","journal":"ACS omega","confidence":"Low","confidence_rationale":"Tier 3 — pharmacological inhibitor study without direct mechanistic mutagenesis, single lab","pmids":["38973855"],"is_preprint":false}],"current_model":"DHX33 is a nucleolar/cytoplasmic DEAH-box RNA helicase with ATPase-coupled DNA/RNA unwinding activity that (1) associates with rDNA loci via UBF to promote RNA Pol I-dependent 47S rRNA transcription and ribosome biogenesis, (2) facilitates elongation-competent 80S ribosome assembly to drive mRNA translation initiation, (3) acts as a transcriptional regulator by binding gene promoters in complex with Gadd45a/Tet1 to cause DNA demethylation and RNA Pol II loading, (4) senses cytosolic dsRNA via its HELICc domain to activate the NLRP3 inflammasome (via NLRP3 interaction) or IPS-1/MAVS-dependent type I IFN signaling in innate immune cells, and (5) is post-translationally regulated by GSK-3β-mediated phosphorylation at T482 (triggering ubiquitin-mediated degradation) and by USP36-mediated deubiquitination (stabilization)."},"narrative":{"teleology":[{"year":2011,"claim":"Establishing that DHX33 is required for rDNA transcription resolved how this DEAH-box helicase participates in ribosome biogenesis: it associates with rDNA loci and UBF to promote RNA Pol I occupancy and 47S rRNA synthesis, with its NTPase activity being essential.","evidence":"RNAi screen, ChIP at rDNA, Co-IP with UBF, dominant-negative K94R mutant, rRNA synthesis assays in human cells","pmids":["21930779"],"confidence":"High","gaps":["Structural basis of DHX33–UBF interaction unknown","Direct versus indirect association with rDNA not resolved","Whether DHX33 unwinds R-loops or other structures at rDNA not tested"]},{"year":2013,"claim":"Identification of DHX33 as a cytosolic dsRNA sensor that activates both the NLRP3 inflammasome and the MAVS-dependent type I IFN pathway revealed a dual innate immune function mediated through its HELICc domain.","evidence":"Domain-mapping Co-IP (HELICc with NLRP3 and IPS-1/MAVS), shRNA knockdown in macrophages and myeloid DCs, caspase-1 and cytokine secretion assays, poly I:C binding assays","pmids":["23871209","24037184"],"confidence":"High","gaps":["Relative contribution of DHX33 versus RIG-I/MDA5 during physiological viral infection not determined","In vivo immune phenotype of DHX33 deletion in myeloid cells not reported"]},{"year":2013,"claim":"Demonstrating that oncogenic Ras upregulates DHX33 translation via PI3K/mTOR/MAPK and that DHX33 is required for Ras-driven rRNA transcription and transformation placed DHX33 as a critical downstream effector linking growth factor signaling to ribosome biogenesis in cancer.","evidence":"Polysome fractionation, kinase inhibitor panels, DHX33 knockdown rescue of RasV12-driven transformation, in vivo tumor model","pmids":["23401854"],"confidence":"High","gaps":["Whether DHX33 translation is directly controlled by mTORC1 phosphorylation of specific factors not shown","ARF-mediated translational repression mechanism not molecularly defined"]},{"year":2015,"claim":"Showing that DHX33 promotes conversion of 80S monosomes to elongation-competent polysomes established a second, translation-level function distinct from its rRNA synthesis role, explaining how DHX33 loss causes global translational inhibition.","evidence":"Polyribosome profiling, RNA immunoprecipitation, Co-IP with ribosomal proteins and translation factors, helicase-dead mutant rescue failure","pmids":["26100019"],"confidence":"High","gaps":["Specific RNA substrates unwound by DHX33 during 80S-to-polysome transition not identified","Whether DHX33 acts on mRNA secondary structures or ribosomal RNA rearrangements unclear"]},{"year":2016,"claim":"Discovery that DHX33 occupies promoters of cell cycle genes and is required for RNA Pol II loading at these loci expanded its role from a ribosome biogenesis factor to a direct transcriptional regulator of Pol II-dependent genes.","evidence":"ChIP for DHX33 and RNA Pol II at cell cycle gene promoters, siRNA knockdown, CRISPR knockout in zebrafish, xenograft tumor model","pmids":["27601587"],"confidence":"High","gaps":["Mechanism by which a helicase facilitates Pol II loading not determined","Whether DHX33 opens chromatin or resolves R-loops at Pol II promoters not tested"]},{"year":2017,"claim":"Identification of USP36 as the deubiquitinase that stabilizes DHX33 revealed the first post-translational regulatory mechanism controlling DHX33 abundance, with loss of USP36 phenocopying DHX33 loss in rRNA synthesis and causing preimplantation lethality in mice.","evidence":"Ubiquitination and protein stability assays, USP36 knockout mouse, Northern blot for rRNA, puromycin incorporation for translation","pmids":["29273634"],"confidence":"High","gaps":["E3 ligase responsible for DHX33 ubiquitination not identified at this point","Specific ubiquitin chain types on DHX33 not characterized"]},{"year":2018,"claim":"Biochemical reconstitution with purified recombinant DHX33 demonstrated that it possesses bona fide ATPase-coupled unwinding activity on both DNA and RNA duplexes, confirming the enzymatic basis of its diverse cellular functions.","evidence":"In vitro ATPase assay with purified recombinant protein, helicase unwinding assay on DNA/RNA substrates, ATP-binding site mutagenesis","pmids":["29870660"],"confidence":"High","gaps":["Substrate specificity (preferred duplex length, sequence, or structure) not systematically characterized","No structural model of DHX33 available"]},{"year":2020,"claim":"Discovery that DHX33 recruits Gadd45a and Tet1 to CG-rich promoters to drive DNA demethylation and histone H4 acetylation provided a mechanistic explanation for how a helicase regulates Pol II transcription—through epigenetic remodeling.","evidence":"ChIP, 5hmC quantification, Co-IP of DHX33–AP-2β–Gadd45a complex, RNA-seq, promoter methylation analysis","pmids":["32312884","32617965"],"confidence":"High","gaps":["Whether R-loop formation by DHX33 is required for Tet1 recruitment not directly shown","Genome-wide map of DHX33 binding sites (ChIP-seq) not reported"]},{"year":2022,"claim":"Identification of GSK-3β-mediated phosphorylation at T482 as a trigger for DHX33 ubiquitination and degradation completed the regulatory circuit, explaining how growth-promoting signals (which inactivate GSK-3β) elevate DHX33 protein in cancer cells; K94 was identified as a major ubiquitination site that also overlaps the ATP-binding region.","evidence":"In vitro kinase assay, T482 mutagenesis, ubiquitination assay, GSK-3β inhibitor/activator treatments, protein stability measurements","pmids":["36403931"],"confidence":"High","gaps":["Identity of the E3 ubiquitin ligase acting downstream of GSK-3β phosphorylation not determined","Whether T482 phosphorylation directly creates a phosphodegron not resolved"]},{"year":2023,"claim":"B-cell-specific Dhx33 deletion in mice demonstrated that DHX33 is dispensable for steady-state B cells but essential for activation-induced rDNA transcription, proliferation, and germinal center responses, providing the first in vivo immune cell-autonomous requirement for DHX33 in ribosome biogenesis.","evidence":"Conditional CRISPR/Cas9 B-cell knockout mouse, 47S rRNA measurement, p53 accumulation assay, germinal center analysis","pmids":["36631557"],"confidence":"High","gaps":["Whether DHX33 loss triggers nucleolar stress in other rapidly proliferating immune cells not examined","Extent to which p53-dependent apoptosis accounts for the full B-cell phenotype not dissected"]},{"year":null,"claim":"Major unresolved questions include the structural basis of DHX33 substrate recognition, the identity of the E3 ligase mediating its phosphodegron-triggered degradation, genome-wide mapping of DHX33 chromatin occupancy, and the physiological relevance of DHX33 in antiviral innate immunity in vivo.","evidence":"","pmids":[],"confidence":"Low","gaps":["No structural model of DHX33","E3 ubiquitin ligase downstream of GSK-3β phosphorylation unknown","No genome-wide ChIP-seq for DHX33 reported","In vivo role in antiviral immunity not tested with conditional knockout models"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140657","term_label":"ATP-dependent activity","supporting_discovery_ids":[0,4,8]},{"term_id":"GO:0003723","term_label":"RNA binding","supporting_discovery_ids":[1,2,4,8,11]},{"term_id":"GO:0003677","term_label":"DNA binding","supporting_discovery_ids":[5,8,12]},{"term_id":"GO:0016787","term_label":"hydrolase activity","supporting_discovery_ids":[8]},{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[5,6,12]},{"term_id":"GO:0140299","term_label":"molecular sensor activity","supporting_discovery_ids":[1,2]}],"localization":[{"term_id":"GO:0005730","term_label":"nucleolus","supporting_discovery_ids":[0,14]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[1,2,11]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[0,5,12]}],"pathway":[{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[0,5,6,12,14]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[4,7,11]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[1,2,14]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[5]},{"term_id":"R-HSA-1852241","term_label":"Organelle biogenesis and maintenance","supporting_discovery_ids":[0,14]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[12,15]}],"complexes":["NLRP3 inflammasome"],"partners":["UBF","NLRP3","MAVS","USP36","GSK3B","GADD45A","TET1","DDX3X"],"other_free_text":[]},"mechanistic_narrative":"DHX33 is a DEAH-box RNA helicase that couples ATPase-driven nucleic acid unwinding to ribosome biogenesis, mRNA translation, transcriptional regulation, and innate immune sensing. Its ATPase activity, stimulated by DNA or RNA duplexes, is essential for promoting RNA Pol I-dependent 47S rRNA synthesis at rDNA loci (via interaction with UBF) and for facilitating elongation-competent 80S ribosome assembly required for global mRNA translation initiation [PMID:21930779, PMID:26100019, PMID:29870660]. DHX33 also functions as a transcriptional regulator by binding CG-rich promoters and recruiting Gadd45a/Tet1 to promote DNA demethylation and RNA Pol II loading at target genes including cell cycle regulators and metabolic enzymes [PMID:27601587, PMID:32312884]. In innate immunity, DHX33 senses cytosolic dsRNA through its HELICc domain and activates both the NLRP3 inflammasome and the IPS-1/MAVS-dependent type I interferon pathway, while its protein stability is controlled by opposing USP36-mediated deubiquitination and GSK-3β phosphorylation-triggered ubiquitin-dependent degradation [PMID:23871209, PMID:24037184, PMID:29273634, PMID:36403931]."},"prefetch_data":{"uniprot":{"accession":"Q9H6R0","full_name":"ATP-dependent RNA helicase DHX33","aliases":["DEAH box protein 33"],"length_aa":707,"mass_kda":78.9,"function":"Implicated in nucleolar organization, ribosome biogenesis, protein synthesis and cytoplasmic dsRNA sensing (By similarity) (PubMed:21930779, PubMed:23871209, PubMed:26100019). Stimulates RNA polymerase I transcription of the 47S precursor rRNA. Associates with ribosomal DNA (rDNA) loci where it is involved in POLR1A recruitment (PubMed:21930779). In the cytoplasm, promotes elongation-competent 80S ribosome assembly at the late stage of mRNA translation initiation (PubMed:26100019). Senses cytosolic dsRNA mediating NLRP3 inflammasome formation in macrophages and type I interferon production in myeloid dendritic cells (PubMed:23871209). Required for NLRP3 activation induced by viral dsRNA and bacterial RNA (PubMed:23871209). In dendritic cells, required for induction of type I interferon production induced by cytoplasmic dsRNA via the activation of MAPK and NF-kappa-B signaling pathways (By similarity)","subcellular_location":"Nucleus, nucleolus; Nucleus, nucleoplasm; Cytoplasm; Nucleus; Inflammasome","url":"https://www.uniprot.org/uniprotkb/Q9H6R0/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/DHX33","classification":"Common Essential","n_dependent_lines":1166,"n_total_lines":1208,"dependency_fraction":0.9652317880794702},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"USP36","stoichiometry":4.0},{"gene":"FKBP5","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/DHX33","total_profiled":1310},"omim":[{"mim_id":"614405","title":"DEAH-BOX HELICASE 33; DHX33","url":"https://www.omim.org/entry/614405"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Enhanced","locations":[{"location":"Nucleoli","reliability":"Enhanced"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/DHX33"},"hgnc":{"alias_symbol":["FLJ21972","DKFZp762F2011"],"prev_symbol":["DDX33"]},"alphafold":{"accession":"Q9H6R0","domains":[{"cath_id":"3.40.50.300","chopping":"64-252","consensus_level":"high","plddt":90.3928,"start":64,"end":252},{"cath_id":"3.40.50.300","chopping":"259-434","consensus_level":"high","plddt":91.0686,"start":259,"end":434},{"cath_id":"1.10.10","chopping":"439-500","consensus_level":"medium","plddt":89.2418,"start":439,"end":500},{"cath_id":"-","chopping":"505-707","consensus_level":"medium","plddt":90.5547,"start":505,"end":707}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9H6R0","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9H6R0-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9H6R0-F1-predicted_aligned_error_v6.png","plddt_mean":85.31},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=DHX33","jax_strain_url":"https://www.jax.org/strain/search?query=DHX33"},"sequence":{"accession":"Q9H6R0","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9H6R0.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9H6R0/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9H6R0"}},"corpus_meta":[{"pmid":"23871209","id":"PMC_23871209","title":"The DHX33 RNA helicase senses cytosolic RNA and activates the NLRP3 inflammasome.","date":"2013","source":"Immunity","url":"https://pubmed.ncbi.nlm.nih.gov/23871209","citation_count":173,"is_preprint":false},{"pmid":"21930779","id":"PMC_21930779","title":"Identification of DHX33 as a mediator of rRNA synthesis and cell growth.","date":"2011","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/21930779","citation_count":55,"is_preprint":false},{"pmid":"24037184","id":"PMC_24037184","title":"The interaction between the helicase DHX33 and IPS-1 as a novel pathway to sense double-stranded RNA and RNA viruses in myeloid dendritic cells.","date":"2013","source":"Cellular & molecular immunology","url":"https://pubmed.ncbi.nlm.nih.gov/24037184","citation_count":49,"is_preprint":false},{"pmid":"29273634","id":"PMC_29273634","title":"Loss of the deubiquitinase USP36 destabilizes the RNA helicase DHX33 and causes preimplantation lethality in mice.","date":"2017","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/29273634","citation_count":38,"is_preprint":false},{"pmid":"27693040","id":"PMC_27693040","title":"miR-634 exhibits anti-tumor activities toward hepatocellular carcinoma via Rab1A and DHX33.","date":"2016","source":"Molecular oncology","url":"https://pubmed.ncbi.nlm.nih.gov/27693040","citation_count":35,"is_preprint":false},{"pmid":"26100019","id":"PMC_26100019","title":"The DHX33 RNA Helicase Promotes mRNA Translation Initiation.","date":"2015","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/26100019","citation_count":30,"is_preprint":false},{"pmid":"28498893","id":"PMC_28498893","title":"Role of DHX33 in c-Myc-induced cancers.","date":"2017","source":"Carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/28498893","citation_count":29,"is_preprint":false},{"pmid":"23401854","id":"PMC_23401854","title":"P19ARF and RasV¹² offer opposing regulation of DHX33 translation to dictate tumor cell fate.","date":"2013","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/23401854","citation_count":24,"is_preprint":false},{"pmid":"27601587","id":"PMC_27601587","title":"DHX33 Transcriptionally Controls Genes Involved in the Cell Cycle.","date":"2016","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/27601587","citation_count":24,"is_preprint":false},{"pmid":"30552990","id":"PMC_30552990","title":"The RNA helicase DHX33 is required for cancer cell proliferation in human glioblastoma and confers resistance to PI3K/mTOR inhibition.","date":"2018","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/30552990","citation_count":24,"is_preprint":false},{"pmid":"32004669","id":"PMC_32004669","title":"DHX33 promotes colon cancer development downstream of Wnt signaling.","date":"2020","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/32004669","citation_count":18,"is_preprint":false},{"pmid":"33843021","id":"PMC_33843021","title":"Long non-coding RNA HOTAIR promotes hepatocellular carcinoma progression by regulating miR-526b-3p/DHX33 axis.","date":"2021","source":"Genes & genomics","url":"https://pubmed.ncbi.nlm.nih.gov/33843021","citation_count":17,"is_preprint":false},{"pmid":"32717723","id":"PMC_32717723","title":"Circular RNA DHX33 promotes malignant behavior in ccRCC by targeting miR-489-3p/MEK1 axis.","date":"2020","source":"Aging","url":"https://pubmed.ncbi.nlm.nih.gov/32717723","citation_count":17,"is_preprint":false},{"pmid":"32617965","id":"PMC_32617965","title":"Function of DHX33 in promoting Warburg effect via regulation of glycolytic genes.","date":"2020","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/32617965","citation_count":16,"is_preprint":false},{"pmid":"32767810","id":"PMC_32767810","title":"Targeting RNA helicase DHX33 blocks Ras-driven lung tumorigenesis in vivo.","date":"2020","source":"Cancer science","url":"https://pubmed.ncbi.nlm.nih.gov/32767810","citation_count":13,"is_preprint":false},{"pmid":"36631557","id":"PMC_36631557","title":"Dhx33 promotes B-cell growth and proliferation by controlling activation-induced rRNA upregulation.","date":"2023","source":"Cellular & molecular immunology","url":"https://pubmed.ncbi.nlm.nih.gov/36631557","citation_count":11,"is_preprint":false},{"pmid":"32312884","id":"PMC_32312884","title":"DHX33 Recruits Gadd45a To Cause DNA Demethylation and Regulates a Subset of Gene Transcription.","date":"2020","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/32312884","citation_count":9,"is_preprint":false},{"pmid":"29870660","id":"PMC_29870660","title":"Recombinant DHX33 Protein Possesses Dual DNA/RNA Helicase Activity.","date":"2018","source":"Biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/29870660","citation_count":8,"is_preprint":false},{"pmid":"23890068","id":"PMC_23890068","title":"RNA helicase DHX33 puts a new twist on NLRP3 inflammasome activation.","date":"2013","source":"Immunity","url":"https://pubmed.ncbi.nlm.nih.gov/23890068","citation_count":7,"is_preprint":false},{"pmid":"36403931","id":"PMC_36403931","title":"GSK-3β phosphorylation of DHX33 leads to its ubiquitination mediated protein degradation.","date":"2022","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/36403931","citation_count":4,"is_preprint":false},{"pmid":"37838340","id":"PMC_37838340","title":"Development of small molecule inhibitors targeting RNA helicase DHX33 as anti-cancer agents.","date":"2023","source":"Bioorganic & medicinal chemistry letters","url":"https://pubmed.ncbi.nlm.nih.gov/37838340","citation_count":3,"is_preprint":false},{"pmid":"38143011","id":"PMC_38143011","title":"DHX33 mediates p53 to regulate mevalonate pathway gene transcription in human cancers.","date":"2023","source":"Biochimica et biophysica acta. General subjects","url":"https://pubmed.ncbi.nlm.nih.gov/38143011","citation_count":2,"is_preprint":false},{"pmid":"29864424","id":"PMC_29864424","title":"Alternative translation initiation from two in-frame start codons in DHX33 gene.","date":"2018","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/29864424","citation_count":2,"is_preprint":false},{"pmid":"30684270","id":"PMC_30684270","title":"A 54-kDa short variant of DHX33 functions in regulating mRNA translation.","date":"2019","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/30684270","citation_count":1,"is_preprint":false},{"pmid":"37543097","id":"PMC_37543097","title":"RNA helicase DHX33 regulates HMGB family genes in human cancer cells.","date":"2023","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/37543097","citation_count":1,"is_preprint":false},{"pmid":"38973855","id":"PMC_38973855","title":"An RNA Helicase DHX33 Inhibitor Shows Broad Anticancer Activity via Inducing Ferroptosis in Cancer Cells.","date":"2024","source":"ACS omega","url":"https://pubmed.ncbi.nlm.nih.gov/38973855","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":10960,"output_tokens":3965,"usd":0.046178},"stage2":{"model":"claude-opus-4-6","input_tokens":7447,"output_tokens":3223,"usd":0.176715},"total_usd":0.222893,"stage1_batch_id":"msgbatch_0118RScg3WGaGkaZRtWZbtce","stage2_batch_id":"msgbatch_01Cpfzvtbj42aqp5oYZ5Te2s","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2011,\n      \"finding\": \"DHX33 is a cell cycle-regulated nucleolar protein that associates with ribosomal DNA (rDNA) loci and interacts with the RNA Pol I transcription factor UBF; DHX33 knockdown decreases Pol I association with rDNA and dramatically reduces 47S rRNA synthesis, while an NTPase-defective mutant (K94R) acts as a dominant negative inhibitor of rRNA synthesis.\",\n      \"method\": \"lentiviral RNAi screen, ChIP, Co-IP, mutagenesis (K94R dominant negative, DNA-binding defective mutant), rRNA synthesis assays\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — multiple orthogonal methods including mutagenesis, ChIP, Co-IP, and functional assays in a single study\",\n      \"pmids\": [\"21930779\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"DHX33 acts as a cytosolic RNA sensor that binds dsRNA via its helicase C domain, interacts with NLRP3, and forms the inflammasome complex following RNA stimulation, leading to caspase-1 activation and IL-1β/IL-18 secretion in human macrophages.\",\n      \"method\": \"shRNA knockdown, binding domain mapping, Co-IP (DHX33–NLRP3 interaction), caspase-1 activity assay, cytokine secretion assay\",\n      \"journal\": \"Immunity\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — domain-mapping pulldown, Co-IP, and functional inflammasome assays; replicated in independent study (PMID:24037184)\",\n      \"pmids\": [\"23871209\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"DHX33 interacts with IPS-1/MAVS in myeloid dendritic cells via the HELICc domain of DHX33 and the C-terminal domain of IPS-1, independently of RIG-I/MDA5, to mediate dsRNA-induced type I IFN production and activation of MAP kinases, NF-κB, and IRF3.\",\n      \"method\": \"shRNA knockdown, domain-mapping Co-IP (HELICc–IPS-1 C-terminal), poly I:C binding assay, signaling pathway analysis\",\n      \"journal\": \"Cellular & molecular immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal domain-mapping Co-IP with functional signaling readouts; consistent with PMID:23871209\",\n      \"pmids\": [\"24037184\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"DHX33 protein translation is regulated oppositely by ARF (which reduces polysome-associated DHX33 mRNA) and RasV12 (which shifts DHX33 mRNA to actively translating polysomes via PI3K/mTOR/MAPK pathways); DHX33 is required downstream of RasV12 for enhanced rRNA transcription and cellular transformation.\",\n      \"method\": \"polysome fractionation, translational reporter assays, kinase inhibitor treatment, DHX33 knockdown rescue experiments, in vitro and in vivo transformation assays\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods including polysome profiling, pharmacological inhibition, and in vivo tumor models\",\n      \"pmids\": [\"23401854\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"DHX33 promotes mRNA translation initiation by facilitating elongation-competent 80S ribosome assembly; DHX33 reduction markedly reduces polyribosome formation and causes global inhibition of mRNA translation, with an accumulation of mRNAs stalled at the 80S ribosome; helicase-defective DHX33 cannot rescue this phenotype.\",\n      \"method\": \"polyribosome profiling, RNA immunoprecipitation (RIP), ribosomal protein/translation factor Co-IP, helicase-dead mutant rescue\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods including polysome profiling, RIP, and mutagenesis with rescue\",\n      \"pmids\": [\"26100019\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"DHX33 physically associates with promoters of cell cycle genes (cyclins, E2F1, CDC, MCM genes) and controls loading of active RNA polymerase II onto these promoters to drive cell cycle progression; CRISPR-mediated knockout in zebrafish confirmed downregulation of these targets in vivo.\",\n      \"method\": \"ChIP, RNA Pol II ChIP, siRNA knockdown, cell cycle analysis, CRISPR/Cas9 knockout in zebrafish, xenograft model\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — ChIP-based mechanism with multiple orthogonal validations including in vivo zebrafish model\",\n      \"pmids\": [\"27601587\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"c-Myc binds to the DHX33 upstream promoter region and stimulates DHX33 transcription; DHX33 in turn promotes transcription of MMP9, MMP14, and PLAU by directly binding to their promoters, thereby promoting cancer cell migration.\",\n      \"method\": \"ChIP (Myc at DHX33 promoter; DHX33 at MMP/PLAU promoters), luciferase reporter, knockdown/rescue, in vivo mouse leukemia model\",\n      \"journal\": \"Carcinogenesis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — ChIP and functional assays from a single lab\",\n      \"pmids\": [\"28498893\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"USP36 deubiquitinase reduces ubiquitination of DHX33 and increases its protein stability; loss of USP36 destabilizes DHX33, impairs rRNA synthesis and protein translation, and causes preimplantation lethality in mice.\",\n      \"method\": \"ubiquitination assay, protein stability assay, USP36 knockout mouse, shRNA knockdown, Northern blot, O-propargyl-puromycin incorporation, electron microscopy\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — in vivo genetic model combined with biochemical ubiquitination assays and multiple orthogonal methods\",\n      \"pmids\": [\"29273634\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Purified recombinant DHX33 protein possesses ATPase activity stimulated by DNA or RNA duplexes, and this ATPase activity is coupled to unwinding of both RNA and DNA duplexes; mutation of a key residue in the ATP-binding site abolishes unwinding activity.\",\n      \"method\": \"in vitro ATPase assay with purified recombinant protein, helicase unwinding assay, ATP-binding site mutagenesis\",\n      \"journal\": \"Biochemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro biochemical reconstitution with mutagenesis of active site\",\n      \"pmids\": [\"29870660\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"DHX33 is required for glioblastoma cell proliferation and migration; overexpression of wild-type DHX33 but not a helicase-dead mutant confers resistance to mTOR inhibitors, indicating the helicase activity is required for this function.\",\n      \"method\": \"DHX33 knockdown (proliferation/migration assays, xenograft), wild-type vs. helicase-dead mutant rescue, mTOR inhibitor treatment\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — mutagenesis with functional rescue in cancer cell context, single lab\",\n      \"pmids\": [\"30552990\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"DHX33 produces two protein isoforms of different size from two in-frame start codons via alternative translation initiation (leaky scanning); both isoforms have similar cellular localization and functions.\",\n      \"method\": \"mutagenesis of start codons, cell line and mouse model analysis, immunoblotting\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic mutagenesis with multiple model validations, single lab\",\n      \"pmids\": [\"29864424\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"A short DHX33 variant (DHX33-2, 534 aa comprising the C-terminal helicase domain) localizes preferentially to the cytoplasm and interacts with DDX3, eIF3, hnRNPs, and poly(A)-binding protein to stimulate translation of a subset of mRNAs involved in cell proliferation.\",\n      \"method\": \"protein immunoprecipitation, RIP-seq, RNA sequencing, subcellular fractionation\",\n      \"journal\": \"Journal of cellular physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 — Co-IP and RIP with functional translation assays, single lab\",\n      \"pmids\": [\"30684270\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"DHX33 recruits Gadd45a and DNA dioxygenase Tet1 to promoters of specific genes (including glycolytic genes LDHA, PDK1, PKM2), causing local DNA demethylation (reduced 5-hydroxymethylcytosine) and enhanced histone H4 acetylation to promote their transcription; this involves DHX33 binding to CG-rich promoter regions, potentially via R-loop formation.\",\n      \"method\": \"ChIP, 5hmC quantification, Co-IP (DHX33–AP-2β–Gadd45a complex), RNA-seq, promoter methylation analysis, DHX33 knockdown\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods (ChIP, 5hmC assay, Co-IP, RNA-seq) establishing a mechanistic epigenetic pathway\",\n      \"pmids\": [\"32312884\", \"32617965\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"GSK-3β directly phosphorylates DHX33 at T482, triggering ubiquitination-mediated protein degradation; a major ubiquitination site was identified at K94 (also critical for ATP binding/helicase activity); cancer cells with frequent GSK-3β inactivation have elevated DHX33 stability.\",\n      \"method\": \"in vitro kinase assay, phosphorylation site mutagenesis (T482), ubiquitination assay, protein stability assay, GSK-3β inhibitor/activator treatment\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — in vitro kinase assay with mutagenesis and ubiquitination assay, single lab but multiple orthogonal methods\",\n      \"pmids\": [\"36403931\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Dhx33 is required for activation-induced upregulation of ribosomal DNA transcription in B cells; B-cell-specific deletion of Dhx33 impairs B-cell growth and proliferation after activation, causing nucleolar stress, p53 accumulation, and cellular death, without affecting steady-state B-cell function.\",\n      \"method\": \"CRISPR/Cas9-mediated B-cell-specific knockout mouse, rDNA transcription assay, 47S rRNA measurement, p53 immunoblot, germinal center analysis\",\n      \"journal\": \"Cellular & molecular immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean conditional knockout in vivo with mechanistic rRNA and p53 pathway readouts\",\n      \"pmids\": [\"36631557\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"DHX33 promotes expression of mevalonate pathway genes downstream of mutant p53 and Ras; in vivo lung tumors carrying mutant p53/KrasG12D show upregulated mevalonate pathway genes that are debilitated upon DHX33 loss.\",\n      \"method\": \"DHX33 knockdown, in vivo KrasG12D/p53-mutant mouse model, gene expression analysis\",\n      \"journal\": \"Biochimica et biophysica acta. General subjects\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vivo genetic epistasis with pathway gene readout, single lab\",\n      \"pmids\": [\"38143011\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"DHX33 promotes expression of lipid metabolism genes FADS1, FADS2, and SCD1, sensitizing cancer cells to ferroptosis; pharmacological inhibition of DHX33 by KY386 induces ferroptosis-mediated cancer cell death.\",\n      \"method\": \"DHX33 inhibitor (KY386) treatment, ferroptosis pathway assays, gene expression analysis, cell viability assays\",\n      \"journal\": \"ACS omega\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — pharmacological inhibitor study without direct mechanistic mutagenesis, single lab\",\n      \"pmids\": [\"38973855\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"DHX33 is a nucleolar/cytoplasmic DEAH-box RNA helicase with ATPase-coupled DNA/RNA unwinding activity that (1) associates with rDNA loci via UBF to promote RNA Pol I-dependent 47S rRNA transcription and ribosome biogenesis, (2) facilitates elongation-competent 80S ribosome assembly to drive mRNA translation initiation, (3) acts as a transcriptional regulator by binding gene promoters in complex with Gadd45a/Tet1 to cause DNA demethylation and RNA Pol II loading, (4) senses cytosolic dsRNA via its HELICc domain to activate the NLRP3 inflammasome (via NLRP3 interaction) or IPS-1/MAVS-dependent type I IFN signaling in innate immune cells, and (5) is post-translationally regulated by GSK-3β-mediated phosphorylation at T482 (triggering ubiquitin-mediated degradation) and by USP36-mediated deubiquitination (stabilization).\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"DHX33 is a DEAH-box RNA helicase that couples ATPase-driven nucleic acid unwinding to ribosome biogenesis, mRNA translation, transcriptional regulation, and innate immune sensing. Its ATPase activity, stimulated by DNA or RNA duplexes, is essential for promoting RNA Pol I-dependent 47S rRNA synthesis at rDNA loci (via interaction with UBF) and for facilitating elongation-competent 80S ribosome assembly required for global mRNA translation initiation [PMID:21930779, PMID:26100019, PMID:29870660]. DHX33 also functions as a transcriptional regulator by binding CG-rich promoters and recruiting Gadd45a/Tet1 to promote DNA demethylation and RNA Pol II loading at target genes including cell cycle regulators and metabolic enzymes [PMID:27601587, PMID:32312884]. In innate immunity, DHX33 senses cytosolic dsRNA through its HELICc domain and activates both the NLRP3 inflammasome and the IPS-1/MAVS-dependent type I interferon pathway, while its protein stability is controlled by opposing USP36-mediated deubiquitination and GSK-3β phosphorylation-triggered ubiquitin-dependent degradation [PMID:23871209, PMID:24037184, PMID:29273634, PMID:36403931].\",\n  \"teleology\": [\n    {\n      \"year\": 2011,\n      \"claim\": \"Establishing that DHX33 is required for rDNA transcription resolved how this DEAH-box helicase participates in ribosome biogenesis: it associates with rDNA loci and UBF to promote RNA Pol I occupancy and 47S rRNA synthesis, with its NTPase activity being essential.\",\n      \"evidence\": \"RNAi screen, ChIP at rDNA, Co-IP with UBF, dominant-negative K94R mutant, rRNA synthesis assays in human cells\",\n      \"pmids\": [\"21930779\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of DHX33–UBF interaction unknown\", \"Direct versus indirect association with rDNA not resolved\", \"Whether DHX33 unwinds R-loops or other structures at rDNA not tested\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Identification of DHX33 as a cytosolic dsRNA sensor that activates both the NLRP3 inflammasome and the MAVS-dependent type I IFN pathway revealed a dual innate immune function mediated through its HELICc domain.\",\n      \"evidence\": \"Domain-mapping Co-IP (HELICc with NLRP3 and IPS-1/MAVS), shRNA knockdown in macrophages and myeloid DCs, caspase-1 and cytokine secretion assays, poly I:C binding assays\",\n      \"pmids\": [\"23871209\", \"24037184\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Relative contribution of DHX33 versus RIG-I/MDA5 during physiological viral infection not determined\", \"In vivo immune phenotype of DHX33 deletion in myeloid cells not reported\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Demonstrating that oncogenic Ras upregulates DHX33 translation via PI3K/mTOR/MAPK and that DHX33 is required for Ras-driven rRNA transcription and transformation placed DHX33 as a critical downstream effector linking growth factor signaling to ribosome biogenesis in cancer.\",\n      \"evidence\": \"Polysome fractionation, kinase inhibitor panels, DHX33 knockdown rescue of RasV12-driven transformation, in vivo tumor model\",\n      \"pmids\": [\"23401854\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether DHX33 translation is directly controlled by mTORC1 phosphorylation of specific factors not shown\", \"ARF-mediated translational repression mechanism not molecularly defined\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Showing that DHX33 promotes conversion of 80S monosomes to elongation-competent polysomes established a second, translation-level function distinct from its rRNA synthesis role, explaining how DHX33 loss causes global translational inhibition.\",\n      \"evidence\": \"Polyribosome profiling, RNA immunoprecipitation, Co-IP with ribosomal proteins and translation factors, helicase-dead mutant rescue failure\",\n      \"pmids\": [\"26100019\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Specific RNA substrates unwound by DHX33 during 80S-to-polysome transition not identified\", \"Whether DHX33 acts on mRNA secondary structures or ribosomal RNA rearrangements unclear\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Discovery that DHX33 occupies promoters of cell cycle genes and is required for RNA Pol II loading at these loci expanded its role from a ribosome biogenesis factor to a direct transcriptional regulator of Pol II-dependent genes.\",\n      \"evidence\": \"ChIP for DHX33 and RNA Pol II at cell cycle gene promoters, siRNA knockdown, CRISPR knockout in zebrafish, xenograft tumor model\",\n      \"pmids\": [\"27601587\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which a helicase facilitates Pol II loading not determined\", \"Whether DHX33 opens chromatin or resolves R-loops at Pol II promoters not tested\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Identification of USP36 as the deubiquitinase that stabilizes DHX33 revealed the first post-translational regulatory mechanism controlling DHX33 abundance, with loss of USP36 phenocopying DHX33 loss in rRNA synthesis and causing preimplantation lethality in mice.\",\n      \"evidence\": \"Ubiquitination and protein stability assays, USP36 knockout mouse, Northern blot for rRNA, puromycin incorporation for translation\",\n      \"pmids\": [\"29273634\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"E3 ligase responsible for DHX33 ubiquitination not identified at this point\", \"Specific ubiquitin chain types on DHX33 not characterized\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Biochemical reconstitution with purified recombinant DHX33 demonstrated that it possesses bona fide ATPase-coupled unwinding activity on both DNA and RNA duplexes, confirming the enzymatic basis of its diverse cellular functions.\",\n      \"evidence\": \"In vitro ATPase assay with purified recombinant protein, helicase unwinding assay on DNA/RNA substrates, ATP-binding site mutagenesis\",\n      \"pmids\": [\"29870660\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Substrate specificity (preferred duplex length, sequence, or structure) not systematically characterized\", \"No structural model of DHX33 available\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Discovery that DHX33 recruits Gadd45a and Tet1 to CG-rich promoters to drive DNA demethylation and histone H4 acetylation provided a mechanistic explanation for how a helicase regulates Pol II transcription—through epigenetic remodeling.\",\n      \"evidence\": \"ChIP, 5hmC quantification, Co-IP of DHX33–AP-2β–Gadd45a complex, RNA-seq, promoter methylation analysis\",\n      \"pmids\": [\"32312884\", \"32617965\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether R-loop formation by DHX33 is required for Tet1 recruitment not directly shown\", \"Genome-wide map of DHX33 binding sites (ChIP-seq) not reported\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Identification of GSK-3β-mediated phosphorylation at T482 as a trigger for DHX33 ubiquitination and degradation completed the regulatory circuit, explaining how growth-promoting signals (which inactivate GSK-3β) elevate DHX33 protein in cancer cells; K94 was identified as a major ubiquitination site that also overlaps the ATP-binding region.\",\n      \"evidence\": \"In vitro kinase assay, T482 mutagenesis, ubiquitination assay, GSK-3β inhibitor/activator treatments, protein stability measurements\",\n      \"pmids\": [\"36403931\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of the E3 ubiquitin ligase acting downstream of GSK-3β phosphorylation not determined\", \"Whether T482 phosphorylation directly creates a phosphodegron not resolved\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"B-cell-specific Dhx33 deletion in mice demonstrated that DHX33 is dispensable for steady-state B cells but essential for activation-induced rDNA transcription, proliferation, and germinal center responses, providing the first in vivo immune cell-autonomous requirement for DHX33 in ribosome biogenesis.\",\n      \"evidence\": \"Conditional CRISPR/Cas9 B-cell knockout mouse, 47S rRNA measurement, p53 accumulation assay, germinal center analysis\",\n      \"pmids\": [\"36631557\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether DHX33 loss triggers nucleolar stress in other rapidly proliferating immune cells not examined\", \"Extent to which p53-dependent apoptosis accounts for the full B-cell phenotype not dissected\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Major unresolved questions include the structural basis of DHX33 substrate recognition, the identity of the E3 ligase mediating its phosphodegron-triggered degradation, genome-wide mapping of DHX33 chromatin occupancy, and the physiological relevance of DHX33 in antiviral innate immunity in vivo.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No structural model of DHX33\", \"E3 ubiquitin ligase downstream of GSK-3β phosphorylation unknown\", \"No genome-wide ChIP-seq for DHX33 reported\", \"In vivo role in antiviral immunity not tested with conditional knockout models\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140657\", \"supporting_discovery_ids\": [0, 4, 8]},\n      {\"term_id\": \"GO:0003723\", \"supporting_discovery_ids\": [1, 2, 4, 8, 11]},\n      {\"term_id\": \"GO:0003677\", \"supporting_discovery_ids\": [5, 8, 12]},\n      {\"term_id\": \"GO:0016787\", \"supporting_discovery_ids\": [8]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [5, 6, 12]},\n      {\"term_id\": \"GO:0140299\", \"supporting_discovery_ids\": [1, 2]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005730\", \"supporting_discovery_ids\": [0, 14]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [1, 2, 11]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [0, 5, 12]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [0, 5, 6, 12, 14]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [4, 7, 11]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [1, 2, 14]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [5]},\n      {\"term_id\": \"R-HSA-1852241\", \"supporting_discovery_ids\": [0, 14]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [12, 15]}\n    ],\n    \"complexes\": [\n      \"NLRP3 inflammasome\"\n    ],\n    \"partners\": [\n      \"UBF\",\n      \"NLRP3\",\n      \"MAVS\",\n      \"USP36\",\n      \"GSK3B\",\n      \"GADD45A\",\n      \"TET1\",\n      \"DDX3X\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}