| 2007 |
Crystal structure of human PUS10 solved at 2.0 Å resolution, revealing a crescent-shaped molecule with two domains: a conserved pseudouridine synthase catalytic domain and a unique N-terminal THUMP-containing domain. Superposition with other Psi synthase catalytic domains identified a conserved active-site residue set, indicating PUS10 employs a similar catalytic mechanism. The active site is in a deep basic cleft with flexible thumb and forefinger loops proposed to stabilize RNA substrate binding. |
X-ray crystallography at 2.0 Å resolution; structural superposition; electrostatic modeling |
Journal of molecular biology |
High |
17900615
|
| 2008 |
Archaeal Pus10 (Methanocaldococcus jannaschii and Pyrococcus furiosus) functions as both a tRNA Ψ54 and Ψ55 synthase in vitro; the two modifications occur independently and show salt-concentration-dependent variation. Unlike bacterial TruB and yeast Pus4, archaeal Pus10 does not require a U54×A58 reverse Hoogsteen base pair or pyrimidine at position 56 for Ψ55 production. |
In vitro pseudouridine synthase assay with recombinant archaeal Pus10 proteins; nearest-neighbor analysis |
RNA (New York, N.Y.) |
High |
18952823
|
| 2012 |
Archaeal Pus10 and the COG1901/TrmY methyltransferase act sequentially in vitro to produce 1-methylpseudouridine (m1Ψ) at position 54 of tRNA: Pus10 first converts U54 to Ψ54, then TrmY methylates Ψ54 in an AdoMet-dependent reaction. Efficient methylation by TrmY requires Ψ55 at position 55; C55 allows only inefficient methylation and a purine at 55 blocks it entirely. |
In vitro reconstitution with purified recombinant Pus10 (M. jannaschii) and TrmY enzymes; tRNA/TΨ-arm substrates; in vivo deletion of COG1901 gene in Haloferax volcanii with complementation |
RNA (New York, N.Y.) |
High |
22274953
|
| 2013 |
Quantitative biochemical analysis of P. furiosus Pus10 showed high affinity for substrate and product tRNA (Kd ~30 nM), with Km ~400 nM and kcat ~0.9 s⁻¹ for Ψ55 formation. Site-directed mutagenesis of the thumb loop reduced catalytic efficiency, and a new catalytic arginine (Arg208) was identified as likely responsible for flipping the target uridine into the active site. The N-terminal THUMP-containing domain contributes to tRNA binding. Data support an induced-fit binding mechanism. |
In vitro pseudouridine synthase assay; fluorescence binding assay; site-directed mutagenesis |
Journal of molecular biology |
High |
23743107
|
| 2013 |
Using homology models based on human PUS10 crystal structure and site-directed mutagenesis of archaeal Pus10, the forefinger loop, a specific Arg, and a Tyr residue were identified as critical for Ψ54 but not Ψ55 synthase activity. A conserved Leu residue, in addition to the catalytic Asp, is essential for both activities. These findings indicate archaeal Pus10 uses two distinct substrate-uridine recognition mechanisms for Ψ54 vs. Ψ55 that share some common features. |
Homology modeling; site-directed mutagenesis; in vitro pseudouridine synthase assay |
RNA (New York, N.Y.) |
High |
23898217
|
| 2017 |
Human PUS10 is predominantly nuclear but translocates to mitochondria early during TRAIL-induced apoptosis via CRM1-mediated nuclear export, concurrent with cytochrome c and SMAC release. Caspase-3 is required for PUS10 translocation; conversely, PUS10 translocation to mitochondria reciprocally amplifies caspase-3 activity through the intrinsic/mitochondrial pathway, creating a feedback amplification loop. p53 is not involved in TRAIL-induced PUS10 movement. |
Immunofluorescence; immunoblotting; CRM1 inhibitor (leptomycin B); caspase-3 inhibition; apoptosis indicators across multiple TRAIL-sensitive and resistant cell lines |
Cell death & disease |
Medium |
28981101
|
| 2018 |
Human PUS10 produces Ψ54 in select tRNAs including tRNALys3 (the HIV reverse transcriptase primer). This activity is restricted to the cytoplasmic isoform; nuclear PUS10 does not have this activity. The recognition sequence GUUCAm1AAUC (positions 53–61, containing 1-methyladenosine) combined with a stable acceptor stem is required for maximum Ψ54 synthase activity. Recombinant human PUS10 can also generate Ψ55 in tRNAs lacking the Ψ54 recognition sequence. |
In vitro pseudouridine synthase assay with recombinant human Pus10 (SF9-derived); nearest-neighbor analysis; subcellular fractionation |
RNA (New York, N.Y.) |
High |
30530625
|
| 2019 |
Human PUS10 directly binds pri-miRNAs and physically interacts with the microprocessor complex (DROSHA-DGCR8), promoting miRNA biogenesis in the nucleus. Depletion of PUS10 causes accumulation of unprocessed pri-miRNAs and reduction of mature miRNAs. This function is independent of PUS10's catalytic pseudouridine synthase activity. Additionally, PUS10 produces pseudouridines at specific positions in cytoplasmic tRNAs (profiled by sequencing). |
RNA immunoprecipitation; Co-immunoprecipitation with DROSHA/DGCR8; siRNA knockdown; pri-miRNA/mature miRNA quantification; Ψ-seq tRNA profiling; catalytic mutant analysis |
Nature chemical biology |
High |
31819270
|
| 2020 |
Mammalian cytoplasmic PUS10 produces both Ψ54 and Ψ55 in tRNAs containing Ψ54Ψ55 (including retroviral primer tRNAs), whereas nuclear TRUB1 produces Ψ55 in most elongator tRNAs and mitochondrial TRUB2 acts on mitochondrial tRNAs. The nuclear isoform of PUS10 is catalytically inactive and specifically binds unmodified U54U55 versions of Ψ54Ψ55-containing tRNAs as well as A54U55 tRNAiMet, inhibiting TRUB1-mediated U55→Ψ55 conversion in the nucleus. This compartmentalization ensures that Ψ54Ψ55-containing tRNAs are modified exclusively by cytoplasmic PUS10. |
Nearest-neighbor analysis with recombinant proteins and subcellular extracts; specific Ψ55 synthase knockdown cells; RNA binding assays with nuclear PUS10 isoform |
RNA (New York, N.Y.) |
High |
33023933
|
| 2023 |
PUS10 promotes the maturation of miR-194-5p in renal cell carcinoma cells, suppressing RCC migration via a PUS10/miR-194-5p/NUDC/Cofilin1 axis. This function is independent of PUS10's classical pseudouridine catalytic activity. HIF-1A transcriptionally represses PUS10 under hypoxic conditions. |
RNA immunoprecipitation; dual luciferase reporter assay; chromatin immunoprecipitation; transwell/wound-healing assay; in vivo metastasis model; catalytic mutant analysis; microRNA sequencing |
Cell & bioscience |
Medium |
37596681
|
| 2025 |
Loss of Pus10 in knockout mice causes cell-intrinsic upregulation of interferon signaling. Mechanistically, Pus10 loss alters the abundance of tRNA-derived small RNAs (tdRs), perturbing translation and endogenous retroelement expression, which promotes accumulation of proinflammatory RNA-DNA hybrids that activate the cGAS-STING pathway. Supplementation with specific tdR pools partly rescues these effects by interacting with RNA processing factors that modulate immune responses. |
Pus10 knockout mouse; IFN signaling readouts; tRNA-derived small RNA profiling; retroelement expression analysis; RNA-DNA hybrid detection; tdR supplementation rescue experiments |
Cell reports |
Medium |
40402745
|