| 1997 |
Yeast Prp43 (DHX15 ortholog) is a pre-mRNA processing factor required for spliceosome disassembly after mRNA release; temperature-sensitive mutations cause accumulation of unspliced pre-mRNAs and excised intron lariats trapped in spliceosomes. |
Gene replacement with ts alleles, Northern analysis, splicing complex analysis |
Proceedings of the National Academy of Sciences of the United States of America |
High |
9342317
|
| 2002 |
Purified recombinant Prp43 is an RNA-dependent ATPase; alanine mutations in motifs I (GSGKT), II (DEAH), and VI (QRAGRAGR) abolish ATPase activity in vitro and are lethal in vivo. An ATPase-dead mutant (T123A) blocks release of the excised lariat-intron from the spliceosome without affecting mRNA release or chemical splicing steps; the lariat-intron in T123A-arrested complexes is inaccessible to debranching enzyme Dbr1. |
Recombinant protein purification, in vitro ATPase assay, active-site mutagenesis, in vitro splicing assay, dominant-negative overexpression |
The Journal of biological chemistry |
High |
11886864
|
| 2002 |
Human DDX15 (DHX15) colocalizes with spliceosomal snRNPs in nuclear speckles and in nucleoli, and is associated with spliceosomal U snRNAs by RNA co-precipitation. DDX15 interacts with the human La autoantigen both in vivo and in vitro; the interaction requires a region that partly overlaps the DEAH-box domain. |
Large-scale immunoprecipitation from HeLa S100 extracts, in vitro binding, immunofluorescence, RNA co-precipitation |
RNA (New York, N.Y.) |
Medium |
12458796
|
| 2005 |
Prp43 forms a stable NTR complex with the novel splicing factors Ntr1 and Ntr2; the affinity-purified NTR complex catalyzes ATP-dependent spliceosome disassembly, releasing U2, U5, U6, the NineTeen Complex, and the lariat-intron. Ntr1 interacts with Prp43 through its N-terminal G-patch domain. |
Immunoprecipitation, in vitro spliceosome disassembly assay with purified NTR complex, domain mapping |
Genes & development |
High |
16357217
|
| 2006 |
Prp43 hydrolyzes all common NTPs/dNTPs and unwinds short 5'/3'-tailed RNA/DNA duplexes in an ATP-dependent manner; optimal ATPase requires RNA cofactor ≥20 nt. Motif V mutations T384A and T384V are lethal and block lariat release despite retaining ATPase activity, demonstrating that ATPase activity is necessary but not sufficient for function. |
In vitro ATPase and helicase unwinding assays, site-directed mutagenesis, in vitro splicing assay |
Biochemistry |
High |
16700561
|
| 2006 |
Ntr1 (Spp382) is required for association of Prp43 with the excised intron complex; depletion of Ntr1 causes accumulation of excised intron and impaired snRNP recycling, indicating Ntr1 acts as a spliceosome receptor or RNA-targeting factor for Prp43. |
Metabolic depletion of Ntr1, in vitro splicing assay, snRNP sedimentation analysis, immunoprecipitation |
Molecular and cellular biology |
High |
16880513
|
| 2007 |
Ntr1 activates the inherently feeble helicase activity of Prp43 to trigger lariat-intron release; the N-terminal 120-aa segment of Ntr1 suffices for Prp43 binding and helicase stimulation. Lethal Prp43 mutants T384A/T384V that retain ATPase activity are refractory to Ntr1-mediated helicase stimulation. Specific missense mutations in Prp43 and Ntr1 that disrupt protein-protein interaction also impair RNA unwinding stimulation. |
In vitro helicase assay, protein-protein interaction mapping, site-directed mutagenesis, in vitro splicing assay |
Genes & development |
High |
17875666
|
| 2007 |
The Ntr1-Ntr2 complex binds the spliceosome before recruiting Prp43; Ntr2 interacts with U5 component Brr2 and is essential for NTR-spliceosome docking. Binding of the NTR complex does not require ATP, but spliceosome disassembly requires ATP hydrolysis. |
Immunoprecipitation, in vitro spliceosome-binding and disassembly assays, two-hybrid and deletion analysis |
Molecular and cellular biology |
High |
17893323
|
| 2009 |
In yeast, functional cooperation between Nob1 (endonuclease) and Prp43 with its cofactor Pfa1 is required for 20S-to-18S pre-rRNA cleavage at site D; genetic epistasis shows that loss of Ltv1 combined with Prp43 or Pfa1 mutation blocks cytoplasmic site D cleavage, which is suppressed by wild-type but not catalytic-dead Nob1. |
Genetic epistasis, in vitro cleavage assay with purified Nob1, pre-rRNA Northern analysis |
The Journal of biological chemistry |
High |
19801658
|
| 2009 |
UV crosslinking identifies multiple Prp43-binding sites on pre-rRNA: predominantly within helix 44 of 18S (near the site of 18S 3' cleavage) and four major sites in 25S including helix 34. Depletion or catalytic inactivation of Prp43 causes accumulation of snoRNAs that guide modifications near helix 34 on preribosomes, indicating Prp43 releases snoRNAs from specific pre-rRNA sites. |
UV crosslinking and immunoprecipitation (CRAC precursor), Northern analysis of snoRNA association, catalytic point-mutant strains |
Molecular cell |
High |
19941819
|
| 2010 |
Prp43 acts as a downstream discard-pathway factor for proofreading 5' splice site cleavage: after Prp16-mediated rejection of suboptimal substrates, Prp43 is required to disassemble the rejected spliceosome. Prp16 and Prp43 thus cooperate in an ATP-dependent fidelity framework. |
In vitro splicing assay using metal-ligand disruption of catalytic center, DEAH-box ATPase inactivation |
Molecular cell |
High |
20705241
|
| 2012 |
Human RBM5 directly interacts with DHX15 via its G-patch domain, and recombinant RBM5 stimulates DHX15 helicase activity in vitro in a G-patch-domain-dependent manner. |
Co-immunoprecipitation, in vitro helicase assay, domain deletion analysis |
FEBS letters |
Medium |
22569250
|
| 2013 |
The minimal activating fragment of Ntr1 is residues 51–110 (containing the G-patch). Cross-linking mass spectrometry shows Prp43 interacts with the G-patch motif of Ntr1 through its C-terminal domains. Functionally important RNA-binding residues were identified in both Prp43 and Ntr1. |
Biochemical ATPase/helicase assays, structural mass spectrometry (protein cross-linking), domain truncation |
Nucleic acids research |
High |
24165877
|
| 2014 |
Human DHX15 physically interacts with MAVS and mediates MAVS-dependent activation of the NF-κB, JNK, and p38 MAPK pathways (but not IRF3) in response to poly(I:C) and RNA virus infection. DHX15 is required for optimal cytokine production and MAVS-mediated apoptosis. |
Co-immunoprecipitation, siRNA knockdown, reporter assays, Drosophila misexpression screen |
Science signaling |
High |
24782566
|
| 2014 |
DHX15 binds double-stranded RNA (poly(I:C)) specifically via its helicase C-terminal domain. The N-terminal DEXDc-containing domain of DHX15 binds the C-terminus of MAVS. DHX15 is required for IRF3 phosphorylation as well as NF-κB and MAPK signaling during RNA virus infection in myeloid dendritic cells. |
RNA pulldown, domain-mapping co-immunoprecipitation, shRNA knockdown, signaling assays |
Journal of immunology |
High |
24990078
|
| 2015 |
The G-patches of Spp382/Ntr1, Sqs1/Pfa1, and Pxr1/Gno1 differ in their ability to interact with and functionally substitute for each other in Prp43-dependent splicing and rRNA processing. Deletion of the primary Prp43-binding site in Pxr1 does not impair rRNA processing but causes accumulation of extended snoRNA forms, linking Prp43 to snoRNA biogenesis. |
Yeast two-hybrid, domain-swap mutagenesis, site-directed mutagenesis, Northern analysis of rRNA and snoRNA |
Genetics |
Medium |
25808954
|
| 2016 |
Crystal structure of Chaetomium thermophilum Prp43 resolved to 2.9 Å; the protein functionally replaces S. cerevisiae Prp43 in spliceosomal disassembly assays. |
X-ray crystallography, in vitro spliceosome disassembly complementation assay |
Acta crystallographica. Section F, Structural biology communications |
High |
26841761
|
| 2017 |
Crystal structures of Prp43 complexes (ATP-analog•RNA, ADP-bound) reveal: RNA sits in a tunnel formed by two RecA-like and C-terminal domains; in the ATP-bound state the tunnel opens into a groove via large C-terminal domain rearrangements; conformational changes between ATP- and ADP-bound states couple ATP hydrolysis to RNA translocation via a β-turn (RecA1 RF motif). The mechanism is distinct from DEAD-box and other helicase families. |
X-ray crystallography of multiple functional states, structure-based mutagenesis |
eLife |
High |
28092261
|
| 2017 |
The stacking of ATP base between R-motif (R159, RecA1) and F-motif (F357, RecA2) residues is required for coupling NTPase and helicase activities in Prp43. F357A mutation or pyrimidine nucleotides decouple NTPase from helicase activity; R159A reduces both. G-patch protein activation is linked to this unique nucleotide-binding mode. |
In vitro ATPase and helicase assays, site-directed mutagenesis, nucleotide analogue substitution |
Nucleic acids research |
High |
28180308
|
| 2017 |
Human DHX15 functions as an AR co-activator by forming a complex with E3 ligase Siah2 and AR (through AR's nuclear export signal, NESAR). DHX15 stabilizes Siah2 and enhances its E3 ubiquitin-ligase activity, resulting in AR activation. This function is independent of DHX15's ATPase activity. |
Yeast mutagenesis screen, co-immunoprecipitation, ubiquitination assay, xenograft tumor model, siRNA knockdown |
Oncogene |
Medium |
28991234
|
| 2017 |
Human NKRF (G-patch protein) forms a pre-ribosomal subcomplex with DHX15 and XRN2. NKRF stimulates DHX15 catalytic activity; this is required for an early pre-rRNA cleavage step (A'). NKRF also recruits XRN2 to nucleolar pre-ribosomal complexes for turnover of excised spacer fragments. NKRF binds transcribed spacer regions of pre-rRNA (by CRAC). |
UV crosslinking and CRAC, co-immunoprecipitation, siRNA depletion with Northern analysis, in vitro helicase stimulation assay |
Nucleic acids research |
High |
28115624
|
| 2018 |
Human CMTr1 (RNA cap1 methyltransferase) binds DHX15 via CMTr1's G-patch domain; DHX15 helicase activity stimulates CMTr1 activity specifically on RNA substrates with highly structured 5' termini, in proportion to structural strength. This is the first demonstrated role for DHX15 in post-transcriptional RNA modification. |
Co-immunoprecipitation, in vitro cap methylation assay with structured vs. unstructured RNA substrates, domain mapping |
Philosophical transactions of the Royal Society of London. Series B, Biological sciences |
Medium |
30397098
|
| 2019 |
DHX15 associates with RIG-I CARDs through its amino terminus and the DHX15–RIG-I complex is recruited to MAVS upon virus infection. DHX15 selectively binds PAMP RNA to promote RIG-I ATP hydrolysis and signaling activation but cannot substitute for RIG-I. DHX15 knockdown increased susceptibility to diverse RNA viruses. |
Co-immunoprecipitation, RNA binding assay, ATPase assay, siRNA knockdown, viral replication assay |
Journal of interferon & cytokine research |
Medium |
31090472
|
| 2020 |
The OB-fold β4-β5 loop of Prp43 is crucial for binding the G-patch of Pfa1 but not for PINX1 binding, revealing distinct binding modes for different G-patch cofactors. Despite different binding modes, stimulation of ATPase and helicase activities by both Pfa1 and PINX1 requires the β4-β5 loop. Disruption of this loop abrogates Prp43 activity in ribosome biogenesis in vivo. |
Mutagenesis, in vitro ATPase/helicase assays, yeast growth and rRNA processing assays |
RNA biology |
High |
32882145
|
| 2021 |
DHX15 interacts with NLRP6 to trigger NLRP6 inflammasome assembly and activation for IL-18 secretion in intestinal epithelial cells (IECs) upon poly(I:C) and enteric RNA virus stimulation. IEC-specific Dhx15 knockout mice show impaired IFN-β, IFN-λ3, and IL-18 production and increased susceptibility to rotavirus and reovirus. |
Conditional knockout mouse model, co-immunoprecipitation, cytokine ELISA, viral infection in vivo |
Cell reports |
High |
34161762
|
| 2021 |
TFIP11 (human Ntr1 homolog) has DHX15-independent roles: it is essential for 2'-O-methylation of U6 snRNA by controlling fibrillarin/snoRNA association, and for U4/U6.U5 tri-snRNP assembly and splicing fidelity. These functions do not require interaction with DHX15. |
TFIP11 knockdown, 2'-O-methylation mapping, snRNP sedimentation, splicing fidelity assays |
Nature communications |
High |
34789764
|
| 2022 |
DHX15 immunodepletion increases A-complex accumulation during in vitro spliceosome assembly and stabilizes an atypical ATP-independent U2 snRNP interaction with a minimal substrate, suggesting DHX15 plays a quality-control role in U2 snRNP engagement with introns. RNase H probing identified nucleotides in the branch-binding region of U2 snRNA that become accessible with GTP hydrolysis (implicating a DEAH enzyme). |
Immunodepletion from splicing extract, in vitro spliceosome assembly assay, RNase H mapping of U2 snRNA |
RNA (New York, N.Y.) |
Medium |
35046126
|
| 2022 |
DHX15 deletion in mouse B cells (conditional KO) impairs lymphocyte development, reduces peripheral B cell numbers, and impairs primary IgG1 responses to immunization, demonstrating an intrinsic requirement for DHX15 in B cell proliferation, survival, and humoral immunity. |
Conditional knockout mice, flow cytometry, antigen immunization, B cell proliferation assay |
Frontiers in immunology |
Medium |
31921164
|
| 2022 |
DHX15 conditional deletion in NK cells reduces NK cell numbers, blocks NK cell maturation, impairs cytolytic function, and abolishes IL-15 responsiveness by inhibiting CD122 surface expression. DHX15 facilitates CD122 surface expression through interaction with CD122 3'UTR in an ATPase-domain-dependent manner, without affecting CD122 mRNA splicing or stability. Ectopic constitutively-active STAT5 rescues the phenotype. |
Conditional NK-cell knockout mice, flow cytometry, cytotoxicity assay, co-immunoprecipitation, mRNA stability assay, rescue with active STAT5 |
Cellular & molecular immunology |
Medium |
35322175
|
| 2022 |
smFRET analysis of Prp43 shows that the G-patch protein Pfa1 induces an open conformation of the RecA domains, accelerating ADP release and enabling transition to the strong RNA-binding apo state. Pfa1 enables Prp43(ADP) to switch between RNA-bound and RNA-unbound states rather than dissociating, making translocation faster than dissociation and enabling processive movement. |
Single-molecule FRET (smFRET), in vitro ATPase and helicase assays, nucleotide-state manipulation |
Proceedings of the National Academy of Sciences of the United States of America |
High |
36409901
|
| 2022 |
DHX15 is identified as the DEAH-box helicase that functions with SUGP1 in splicing. Crystal structure of the human DHX15–SUGP1 G-patch complex reveals the molecular basis of direct interaction. DHX15 depletion or expression of AML-associated DHX15 mutants partially recapitulates the missplicing pattern of mutant SF3B1 cancers. A DHX15–SUGP1 G-patch fusion rescues SF3B1-mutant splicing defects when incorporated into the spliceosome. |
Crystal structure, protein-protein interaction assays (multiple), DHX15 depletion, mutant expression, splicing reporter assays, spliceosome incorporation assay |
Proceedings of the National Academy of Sciences of the United States of America |
High |
36459648
|
| 2023 |
DHX15 promotes splicing quality control (QC) of suboptimal introns with weak splice sites, multiple branch points, and cryptic introns in human cells; this QC function requires DHX15's ATPase activity. SUGP1 is the G-patch factor that recruits and activates DHX15 for splicing QC, dependent on both DHX15 ATPase activity and SUGP1's ULM domain. |
Rapid protein depletion (degron system), nascent and mature RNA sequencing, domain mutant analysis |
Cell reports |
High |
37805921
|
| 2023 |
DHX15 interacts with MYC protein directly (co-localization in cells and direct in vitro interaction). DHX15 stabilizes MYC at the post-translational level by interfering with the FBXW7–MYC interaction, thereby preventing MYC polyubiquitylation and proteasomal degradation. This function is independent of DHX15's RNA-binding capacity. |
Proteomic interactome analysis, co-immunoprecipitation, in vitro binding, MYC stability assay, ubiquitination assay, rescue overexpression of MYC |
iScience |
Medium |
38161423
|
| 2023 |
DHX15 abrogation in T-ALL perturbs RNA splicing and causes intron retention in SLC7A6 and SLC38A5, reducing glutamine import and suppressing mTORC1 activity, thereby blocking T cell development at the DN-to-DP transition and impairing leukemia cell survival. |
Multiple murine T-ALL models, single-cell transcriptomics, splicing analysis, metabolic assay, mTORC1 pathway readouts |
Haematologica |
Medium |
36861414
|
| 2023 |
smFRET analysis shows the RNA binding channel of Prp43 alternates between open and closed conformations. Binding of Pfa1 G-patch and ATP shifts the channel to the open state, facilitating RNA loading. Once RNA is loaded, the channel remains firmly closed during successive ATP hydrolysis cycles, ensuring stable contact and processive translocation. |
Single-molecule FRET with fluorescent reporters on RNA binding channel, nucleotide-state manipulation |
Nucleic acids research |
High |
37167006
|
| 2024 |
GPATCH4 is a stimulatory G-patch cofactor of DHX15 that interacts with DHX15 in the nucleolus via its G-patch domain. GPATCH4 associates with pre-ribosomal particles and crosslinks to pre-rRNA and snoRNAs. Loss of GPATCH4 impairs 2'-O-methylation at multiple rRNA and snRNA sites; DHX15 ATPase activity is required for methylation at DHX15-dependent sites, suggesting DHX15 regulates snoRNA-guided 2'-O-methylation. |
Co-immunoprecipitation, UV crosslinking, 2'-O-methylation mapping (RiboMeth-seq), ATPase-dead mutant analysis, siRNA depletion |
Nucleic acids research |
High |
38113271
|
| 2024 |
Cryo-EM and hydrogen-deuterium exchange MS reveal that G-patch activators Tma23 and Pxr1 each contain an inhibitory segment (I-patch) that binds the catalytic RecA-like domains of Prp43 and allosterically restrains its ATPase activity; both also contain dimerization segments that organize Prp43 into higher-order complexes. Toggling between I-patch inhibition and G-patch activation is proposed to coordinate Prp43 function at discrete pre-rRNA sites. |
Cryo-electron microscopy, hydrogen-deuterium exchange mass spectrometry, in vitro ATPase assays |
Nature communications |
High |
39578461
|
| 2024 |
Mason-Pfizer monkey virus (M-PMV) recruits DHX15 from the nucleus to cytoplasmic viral inclusion bodies via mimicry of the G-patch interaction mechanism, and this recruitment is essential for correct packaging of the viral genome and viral infectivity. |
Genetic engineering, viral genome packaging assay, infectivity assay, localization imaging |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
39320912
|
| 2024 |
DHX15 interacts with mouse APOBEC3 and inhibits its cytidine deaminase activity. DHX15 knockdown inhibits murine leukemia virus (MLV) replication and results in more G-to-A mutations in proviral DNA; DHX15 knockdown also induces DNA damage, implicating DHX15 in genome integrity maintenance in cells expressing mouse APOBEC3. |
Co-immunoprecipitation, deaminase activity assay, DHX15 siRNA knockdown, viral replication assay, proviral mutation analysis |
PLoS pathogens |
Medium |
40168451
|
| 2024 |
F. nucleatum invades intestinal epithelial cells and physically binds to DHX15; knockout of Dhx15 in Villin-Cre/KrasG12D+/- mice attenuates colorectal cancer progression, linking DHX15 to F. nucleatum-dependent oncogenic ERK/STAT3 signaling. |
Co-immunoprecipitation/binding assay, Dhx15 conditional KO mouse model, tumor progression analysis |
Nature communications |
Medium |
38402201
|
| 2025 |
GPATCH3 interacts with DHX15 and enhances its ATPase activity, promoting spliceosome disassembly. Loss of GPATCH3 leads to splicing alterations in immunoregulatory genes (CXCR3, CD44, FOXP3) and reduced tumor growth in vivo. |
Biochemical interaction assay, ATPase activity assay, splicing reporter assays, transcriptomic analysis, in vivo xenograft |
Frontiers in immunology |
Medium |
40861452
|