DDX41 source and assay guide

This guide separates the imported annotation, the observation inspected, and the scope of the inference. Normal publication caches retain their original bytes and access flags. A false full_text_available value in a cache does not mean that a separately recovered original was unavailable.

Identity and products

Original sources and stable locators

Source Actual inspected evidence Access and scope
11991638 Original Results complex assembly and Table 1, “Proteins found only in C complex”: Abstrakt/Q9UJV9; HeLa extract, AdML AG-to-GG substrate, native C-complex purification. Indexed original copy yielded the target table and methods window. Complete original download was not recovered. No specific DDX41 catalytic reaction follows from proteomic membership.
21892174 Original, DNA-binding experiments and ER/localization sections. Human THP1 and mouse dendritic-cell observations are separated; recombinant binding and deletion experiments are distinguished from cellular necessity. Exact construct species is not guessed from host cells.
23142775 Original, cyclic-dinucleotide binding, competition and cellular-response experiments. Mouse D2SC/BMDC/macrophage and human THP1/PBMC systems are distinct. Supplemental construct details not recovered are left unresolved.
22365833 Original spliceosome-interaction screening methods; MINT target pair records ST3/ST6. Human DDX41–NKAP two-hybrid and HEK293 anti-tag co-IP; association does not demonstrate DDX41 transcriptional repression.
22658674 Original page 1394, “In Vivo Capture of HeLa RBPs,” and supplementary methods. Original plus supplement describes UV crosslinking, stringent oligo(dT) capture, RNase release and MS. Separate Table S1 was not recovered: target assignment is deferred to the curator, with independent CLIP corroboration.
23222971 Original DDX41–TRIM21 association and expressed-protein localization experiments. Tagged human-cell cytoplasmic colocalization does not independently resolve a soluble cytosolic fraction. TRIM21 performs ubiquitination.
25416956 Original Results describing pairwise quadruplicate testing from fresh yeast colonies. Identity-confirmed pairs were retained after at least three positive attempts. The literal SIAH1 and CEP70 records were assessed; orthogonal validation of the study set is not claimed for every individual pair.
25920683 Original human splicing/association experiments and hematopoietic phenotypes. Human K562/CD34+/U937/HEK293 evidence and mouse LSK observations are kept distinct. Neither RNA association nor perturbation directly measures RNA helicase activity.
26030138 Results “Validation of CSB interacting proteins by reciprocal co-immunoprecipitation,” and Fig. 5. DDX41–CSB co-precipitation occurs with anti-Myc but not anti-FLAG in human CSIAN lysates. The weak-interaction negative-control example for SMARCA2 is not misdescribed as a DDX41-specific control.
26712909 Original full-length/localization and M1I smaller-product experiments. Full-length WT/R525H/R164W nuclear localization is distinguished from unresolved alternative initiation.
28514442 “Clone Construction and Cell Culture,” “Affinity-Purification Mass Spectrometry,” and background filtering. C-terminal HA-FLAG constructs and cellular co-purification; the NKAP–DDX41 record is HEK293T.
32296183 “Generation and characterization of HuRI”: quadruplicate retesting, sequence confirmation and three assay configurations. NKAPL and AIDA are exact positive pairs; the record does not establish DDX41's native physiological role with either partner.
32703943 Original FAM50A/DDX41 co-IP and cloning methods. Sequence-confirmed full-length V5-DDX41 with endogenous FAM50A in U-87 MG cells; absence of a fresh MITAB match does not erase the original experiment.
32814053 Original Results interaction-mating workflow and “Automated Y2H interaction screening,” ending before “PPI detection in cells using DULIP and LuTHy.” Complete original PDF recovered; literal HTT fragment features remain in the MITAB records. Native neuronal complexes and full-length HTT binding were not established by these records.
33961781 Original construct, AP-MS and CompPASS-Plus methods. Separate HEK293T/HCT116 target records preserve bait orientation. Co-complex recovery is not purified binary binding.
33650667 Independently reviewed human expression and alternative-splicing experiments. Perturbation does not identify an RNA-duplex-unwinding reaction.
34473945 Independently reviewed snoRNA-associated RNA processing, human fusion assays and mouse hematopoietic alleles. HyperTRIBE fusion/replication limits remain; downstream pseudouridylation and protein-synthesis effects do not assign those catalytic reactions to DDX41.
35613581 Original Fig. 3 substrate assays; Fig. 4M nuclear/cytosol fractionation; cGAS/STING response experiments. Tested RNA duplexes are negative, DNA unwinding is ATP dependent, and annealing is ATP independent. Intracellular remodeling balance is a model; cGAS makes cGAMP.
36229594 Independently reviewed K562 UV-CLIP, spliceosome/transcription-factor associations and elongation experiments. RNA binding near 5-prime splice sites does not by itself prove mature-mRNA-only binding; no purified RNA-duplex unwinding was measured.

The original target table for PMID:22681889 and localization assay for PMID:25704810 remain unavailable; their RNA-binding and cytoplasmic-location claims are accepted with independent target evidence and curator deference. PMID:10607561 and PMID:19946888 retain unresolved annotation judgments after bounded recovery attempts. PMID:36780110 was assessed only at abstract level and is not used to assign a direct translation activity.

Literal interaction records and partner classes

All row numbers below are zero-based imported annotation indices. Records were selected by target, partner and PMID, with both human identities and sequence/taxon checksums checked. The multiple records of one pair describe assay stages or contexts, not independent physiological mechanisms.

Rows Pair and literal anchor Interpretation
12 DDX41–NKAP; MINT-8337673 / MINT-8331403 Two-hybrid and anti-tag co-IP; GO:0001222 transcription corepressor binding.
13 DDX41–SIAH1; EBI-10581902 plus pooling/array records GO:0031625 ubiquitin protein ligase binding.
14 DDX41–CEP70; EBI-10592580 plus pooling/array records Positive non-core binding; no DDX41 centrosomal or microtubule mechanism inferred.
15 DDX41–ERCC6/CSB; original Fig. 5 GO:0051117 ATPase binding; asymmetric co-IP retained.
16 NKAP–DDX41; EBI-21520423 HEK293T AP-MS, tagged NKAP; GO:0001222.
17 NKAPL–DDX41; EBI-24640582 and four stage/configuration records Non-core binding; human NKAPL's repression function is assigned by similarity, distinct from the directly supported NKAP class.
18 AIDA–DDX41; EBI-25270277 plus pooling/array records Non-core binding without a DDX41 adaptor or signaling mechanism.
19 FAM50A–DDX41; original human-cell co-IP Non-core association; no catalytic activity inferred.
20 HTT–DDX41; EBI-25957757, EBI-26126967, EBI-26112095, EBI-25972744, EBI-26281098, EBI-26266228 HTT regions 1–511 and 1932–2642; polyglutamine/mutation features retained in source records.
21 DDX41–NKAP EBI-54702916; NKAP–DDX41 EBI-54932614 HEK293T/HCT116 AP-MS respectively; GO:0001222.

The binding refinements use the actual NKAP FUNCTION record, SIAH1 catalytic record, ERCC6 catalytic record, and current definitions of GO:0001222, GO:0031625 and GO:0051117. The current human NKAP FUNCTION record explicitly states “Plays a role as a transcriptional corepressor” and cites PMID:19409814(https://pubmed.ncbi.nlm.nih.gov/19409814/) for its Notch-associated role. This is an experimentally curated partner-class assertion, not a claim that DDX41 mediates repression. GO:0001222 describes binding to a corepressor; it does not require that the observed interaction occur during transcription. Partner-function full originals were not separately reread for this class assessment. The generic thioester-E3 wording in SIAH1's FUNCTION prose is not used as a RING-ligase mechanism.

Propagation and localization

The actual target lineage runs through mRNA-binding node PTN002776867, RNA-helicase node PTN002774595 and DDX41-related spliceosomal node PTN002776089 to leaf PTN002776112. Current IBD rows and donor experimental annotations were inspected. The RNA-helicase question concerns target substrate specificity, not weak donor count or illegitimate self-reference. No residue-loss argument was made.

The literal InterPro rules are IPR011545 to nucleic-acid/ATP binding, IPR014014 to RNA helicase, and IPR044113 to RNA helicase/ATP binding/splicing. Their individual assertions are assessed separately. RHEA:13065 supplies ATP hydrolysis, which agrees with target biochemistry.

HPA reports nucleoplasm as the main localization and nucleoli as an additional localization. Reactome human entity R-HSA-9769601 resolves to Q9UJV9 in nucleoplasm and is explicitly included in C-star/P complexes and release events. R-HSA-3134807 resolves to Q9UJV9 in cytosol and appears in DNA/cyclic-dinucleotide binding and TRIM21 ubiquitination events. These are localization assertions; no other component's catalytic activity is transferred.

Review policy and research provenance

The standing project instruction retains supported positive binding as non-core where a justified more informative term is unavailable. Broad terms with justified replacements use MODIFY even when another source row already carries that term. No new process or molecular-function row was required for the two cores; RNA binding, splicing, DNA-helicase refinement and cGAS/STING participation already have reviewed source coverage.

Normal fetch and PMID retrieval completed. Falcon authentication and its configured fallback's quota failed terminally, so manual primary-source work supplied the review and no provider-named report was fabricated. Original machine caches were preserved, and two additional primary sources, 21892174 and 23142775, were fetched through the normal CLI. Six short cache-verbatim quotations now anchor the substrate correction, ATP hydrolysis, cytosolic fractionation and RNA/splicing functions. Each appears once in the YAML. The exact NKAP FUNCTION quotation above provides the separate partner-class evidence.

The nuclear core retains the encompassing spliceosomal-complex term because the evidence spans C-complex identification and activated-spliceosome association. The more specific catalytic step 2 spliceosome annotation remains accepted; the broader core summary does not assert that every observed DDX41 spliceosome association is restricted to that single stage.