| 2001 |
The WD40 domain of PLRG1 directly interacts with the carboxyl-terminal region of CDC5L in vitro and in vivo. A bacterially expressed CDC5L fragment containing the PLRG1-interaction domain disrupts the CDC5L-PLRG1 interaction in HeLa nuclear extract and inhibits pre-mRNA splicing, demonstrating that this direct interaction is essential for pre-mRNA splicing progression. |
In vitro binding assay, co-immunoprecipitation, domain mapping by mutagenesis, in vitro splicing inhibition assay |
The Journal of biological chemistry |
High |
11544257
|
| 2003 |
Peptides derived from the WD40-containing interaction domain of PLRG1 and the C-terminal domain of CDC5L inhibit pre-mRNA splicing in vitro; this inhibition is rescued by pre-incubating the peptides with the corresponding partner protein, confirming that the CDC5L–PLRG1 interaction is essential for the splicing mechanism. |
In vitro splicing assay with competing peptides, rescue by recombinant protein pre-incubation |
Nucleic acids research |
High |
14576297
|
| 2005 |
PLRG1 is a core component of the Pso4/Prp19–Cdc5L–Plrg1–Spf27 complex (Pso4 complex) that is required for processing of psoralen DNA interstrand cross-links (ICLs) in a cell-free biochemical assay; the complex participates in an early stage of ICL repair alongside MutSβ, Ercc1-Xpf, RPA, and PCNA. |
Cell-free ICL processing assay, immunodepletion, co-immunoprecipitation showing Pso4 complex–WRN association |
The Journal of biological chemistry |
Medium |
16223718
|
| 2007 |
The hPrp19 core complex normally includes Cdc5L, Plrg1, and Spf27; upon DNA damage, hPrp19 forms a ubiquitylated oligomeric species that fails to interact with either Cdc5L or Plrg1, indicating that DNA damage induces structural alterations that disrupt the PLRG1-containing core complex. |
SDS-PAGE under non-reducing conditions, co-immunoprecipitation, chromatin fractionation after DNA damage treatment |
Biochemical and biophysical research communications |
Medium |
17276391
|
| 2009 |
PLRG1 is required for S-phase progression and suppression of apoptosis in vivo. PLRG1-deficient MEFs fail to progress through S phase and show increased apoptosis that is p53-dependent; PLRG1 deficiency causes enhanced p53 phosphorylation/stabilization and increased γ-H2AX, indicating an activated DNA damage response. PLRG1 is also required for nuclear retention of its binding partner CDC5L. |
Conditional knockout mice (heart- and neuron-specific), PLRG1-deficient MEFs, BrdU/FACS cell-cycle analysis, immunofluorescence for γ-H2AX, p53 knockdown rescue in MEFs and zebrafish, subcellular fractionation for CDC5L localization |
Molecular and cellular biology |
High |
19307306
|
| 2009 |
PLRG1 is a core component of the Prp19/Pso4–Cdc5L–Plrg1–Spf27 complex; Cdc5L within this complex interacts physically with ATR kinase and is required for ATR-mediated S-phase checkpoint signaling (Chk1, Rad17, FancD2 activation). PLRG1's role is inferred through its membership in the Cdc5L complex that mediates this checkpoint. |
Co-immunoprecipitation (Cdc5L–ATR), siRNA depletion of Cdc5L, checkpoint kinase activation assays, deletion mapping of ATR-binding region in Cdc5L |
EMBO reports |
Low |
19633697
|
| 2010 |
PLRG1 directly interacts with CDC5L in vivo; a central region of hnRNP-M mediates direct interaction with both CDC5L and PLRG1. This interaction is inhibited during heat-shock stress. An hnRNP-M mutant lacking the CDC5L/PLRG1 interaction domain cannot modulate alternative 5′ and 3′ splice site choice, placing PLRG1 in the pathway of alternative splicing regulation via hnRNP-M. |
Co-immunoprecipitation in vivo, domain mapping by deletion mutants, adeno-E1A minigene alternative splicing assay |
EMBO reports |
Medium |
20467437
|
| 2014 |
PLRG1 (as part of the PSO4 core complex) is required for efficient recruitment of ATRIP to DNA damage sites and subsequent CHK1 activation and RPA2 phosphorylation; PLRG1 colocalizes with RPA at damage sites. Both the RPA1-binding ability of BCAS2 and the E3 ligase activity of PSO4 within the PLRG1-containing complex are required for these functions. |
siRNA depletion of complex subunits, immunofluorescence colocalization, co-immunoprecipitation of PLRG1-complex with RPA, in vitro ATR activation assays |
The Journal of biological chemistry |
Medium |
24443570
|
| 2018 |
PLRG1 is essential for activation of the Prp19 (NTC) E3 ubiquitin ligase. Prp19 is autoinhibited on its own; stepwise assembly of SPF27, CDC5L, and PLRG1 onto the Prp19 tetramer enables ubiquitin ligation activity. Cross-linking/MS and functional assays defined the communication between PLRG1 and Prp19 that enables E3 activity, and crystal structure revealed the autoinhibition mechanism. |
X-ray crystallography of Prp19, mutational analysis of autoinhibition, stepwise reconstitution of NTC core, in vitro ubiquitin ligation assay, protein-protein crosslinking coupled to mass spectrometry |
Molecular cell |
High |
29547724
|
| 2020 |
The crystal structure of the WD40 domain of human PLRG1 was solved by X-ray crystallography. Comparison with cryo-EM structures of PLRG1 within the spliceosome showed that two loops of the WD40 domain become resolved upon binding to other splicing factors, revealing dynamic conformational changes during spliceosome assembly. |
X-ray crystallography (apo WD40 domain), comparison with cryo-EM spliceosome structures |
Biochemical and biophysical research communications |
Medium |
33239170
|
| 2021 |
USP42 (a deubiquitylase) directs the integration of PLRG1 into nuclear speckles (SC35-positive) in a phase-separation-dependent manner; depletion of USP42 displaces PLRG1 from nuclear speckles and deregulates mRNA splicing events phenocopying PLRG1 repression, placing USP42-mediated deubiquitylation upstream of PLRG1 nuclear speckle localization and splicing function. |
Immunofluorescence colocalization, USP42 siRNA knockdown, mRNA splicing assays, phase separation assays, subcellular fractionation |
Cell death and differentiation |
Medium |
33731873
|
| 2022 |
DHX37 interacts with PLRG1 and together they co-occupy the promoter and superenhancer elements of cyclin D1 (CCND1) to transcriptionally activate CCND1 expression, promoting liver cancer cell proliferation. This reveals a non-canonical transcriptional/epigenomic function for PLRG1 beyond splicing. |
Co-immunoprecipitation (DHX37–PLRG1), ChIP-seq epigenomic profiling of DHX37-knockdown cells, ChIP showing co-occupancy at CCND1 promoter/superenhancer, siRNA knockdown proliferation assays |
Cancer research |
Medium |
35290436
|
| 2023 |
PLRG1 knockdown in cancer cells (but not normal cells) causes mitotic arrest, microtubule instability, ER stress, autophagy accumulation, DNA damage, and ultimately apoptosis; in normal cells PLRG1 depletion induces G1 arrest as a self-protective mechanism, distinguishing tumor-specific from normal cell responses to PLRG1 loss. |
siRNA knockdown, cell-cycle FACS analysis, immunofluorescence for microtubule stability and DNA damage markers (γ-H2AX), apoptosis assays |
BMB reports |
Medium |
37817442
|
| 2024 |
YBX1 directly binds to the PLRG1 promoter and transcriptionally activates PLRG1 expression; overexpression of YBX1 upregulates PLRG1 and promotes EMT (increased N-cadherin, Snail, migration, invasion) in HCC cells, and these effects are abolished by PLRG1 knockdown, placing PLRG1 downstream of YBX1 in the EMT pathway. |
Chromatin immunoprecipitation (ChIP) of YBX1 at PLRG1 promoter, luciferase reporter assay, siRNA knockdown of PLRG1, YBX1 overexpression, EMT marker immunoblotting, migration/invasion assays |
Medical oncology |
Medium |
39400789
|