| 2001 |
CSN4 is a component of a smaller cytoplasmic COP9 signalosome subcomplex (~100 kDa, containing CSN4-8 but not CSN1-3) that is distinct from the nuclear 450-kDa holocomplex; this subcomplex arises from nuclear export and is sensitive to leptomycin B treatment. |
Glycerol gradient sedimentation and cell fractionation experiments; leptomycin B treatment |
The Journal of biological chemistry |
Medium |
11704659
|
| 2001 |
CSN4 participates in a pairwise interaction network within the COP9 signalosome; yeast two-hybrid analysis detected Csn4/5/7/6 paired interactions in Arabidopsis, implying a similar quaternary arrangement as in the 26S proteasome lid. |
Yeast two-hybrid analysis of all possible paired subunit interactions |
The EMBO journal |
Low |
11742986
|
| 2002 |
In Drosophila, CSN4 (Drosophila ortholog) forms a complex of similar size to plant and mammalian CSN; null mutations in csn4 are larval lethal and cause defective oocyte/embryo patterning and defects in response to DNA damage, with unique phenotypes reminiscent of ecdysone signaling defects distinct from csn5 mutants. |
Gel filtration with CSN subunit antibodies; null mutant generation and phenotypic characterization |
Development (Cambridge, England) |
Medium |
12223399
|
| 2002 |
In fission yeast, Csn4 is required for removal of Nedd8 from the cullin Pcu1, and its protein product associates with Csn1 and Csn2; however, csn4 null mutants do not share the DNA damage sensitivity and slow replication phenotypes of csn1/csn2 mutants, indicating subunit-specific functional differences. |
Null mutant characterization; co-immunoprecipitation; Nedd8 removal assays |
Molecular biology of the cell |
Medium |
11854407
|
| 2003 |
In Drosophila oogenesis, CSN4 (with CSN5/Jab1) acts through the CSN to remove Nedd8 from Cullin1 (a subunit of SCF ubiquitin ligase), and genetic epistasis shows that Cyclin E is the major downstream target; CSN4 and Cyclin E mutations reciprocally suppress each other. |
Drosophila genetics (CSN4 and CSN5 mutants), genetic epistasis/suppressor analysis, Cyclin E overexpression |
Developmental cell |
Medium |
12737805
|
| 2007 |
The Drosophila CSN4 subunit co-occupies Retinoblastoma target gene promoters with Rbf1 and Rbf2, and physically interacts with Rbf2 during embryogenesis; knockdown of CSN4 leads to increased proteasome-mediated destruction of Rbf1 and Rbf2 and altered cell cycle progression. |
Co-immunoprecipitation; chromatin immunoprecipitation (ChIP); targeted RNAi knockdown with Western blot |
Molecular biology of the cell |
Medium |
17251548
|
| 2009 |
Mass spectrometry-based interaction mapping of reconstituted human CSN identified two symmetrical modules: Csn1/2/3/8 and Csn4/5/6/7, connected by Csn1-Csn6 interactions; the active complex contains a single copy of each of the eight subunits. |
Native mass spectrometry of in vitro reconstituted human CSN complex; subcomplex dissociation mapping |
Structure (London, England : 1993) |
High |
19141280
|
| 2009 |
CSN4 null mutations in Drosophila prevent normal light-dependent Timeless (TIM) degradation in pacemaker neurons and impair behavioral phase shifts; this places CSN4/CSN in the Jetlag (JET) F-box protein pathway for light-mediated TIM degradation. |
Drosophila genetic null mutants; immunofluorescence of TIM in lateral neurons; behavioral phase-shift assays; genetic epistasis with jetlag mutants |
The Journal of neuroscience |
Medium |
19176824
|
| 2009 |
Systematic mass spectrometric pulldown of mammalian CSN4 (among other subunits) defined the protein interaction network of the CSN, identifying stable interactions with a subset of CRL complexes including CRL4(Ddb2), and revealing Dda1 as a new CRL4-associated protein. |
Mass spectrometry-based interactome mapping of CSN subunits including Csn4 by affinity purification |
Journal of cell science |
Medium |
19295130
|
| 2010 |
Human CSN4 (or CSN5) knockdown induces proteasome-mediated degradation of the ubiquitin-conjugating enzyme UBC3/Cdc34 via SCF(betaTrCP); the CSN normally protects UBC3 from ubiquitination requiring UBC3's acidic C-terminal extension. |
siRNA knockdown of CSN4/CSN5; co-immunoprecipitation; in vitro ubiquitination assay; domain mapping |
The Journal of biological chemistry |
Medium |
20378537
|
| 2010 |
In Neurospora crassa, CSN-4 knockout abolishes deneddylation of cullin proteins (Cul1, Cul3, Cul4), destabilizes Cul1 in SCF complexes and Cul3/BTB proteins in Cul3-BTB E3s, and SKP-1 in SCF complexes; this results in severe defects in growth, conidiation, and circadian rhythm. |
Gene knockout mutant generation; neddylation state analysis by immunoblot; phenotypic characterization |
PLoS genetics |
Medium |
21151958
|
| 2010 |
TorsinA (TA) binds directly to CSN4 in neuroblastoma cells and brain synaptosomes; CSN4 and TA are both required for the stability of synaptic proteins snapin and stonin 2; snapin stability is regulated through CSN-associated kinase PKD (protein kinase D) phosphorylation, while stonin 2 stability is regulated through neddylation. |
Co-immunoprecipitation; siRNA knockdown; Western blotting; synaptosome fractionation |
The EMBO journal |
Medium |
21102408
|
| 2012 |
A CSN4-5-6-7 subcomplex was reconstituted in vitro: CSN7, CSN4, and CSN6 form a stable heterotrimer (requiring co-expression); CSN5 can be added to reconstitute the quaternary complex. The subcomplex is stabilized by MPN-MPN interactions (CSN5-CSN6), PCI-PCI interactions (CSN4-CSN7), and CSN6 C-terminus interactions with CSN4 and CSN7; CSN8 also interacts with the CSN4-6-7 core. |
Bacterial co-expression reconstitution; pairwise and combinatorial interaction analysis; biochemical and biophysical characterization |
The Journal of biological chemistry |
High |
23086934
|
| 2014 |
CSN4 was identified as a novel binding partner of soluble guanylyl cyclase α1 (sGCα1) in prostate cancer cells; the CSN4-sGCα1 interaction inhibits sGCα1 proteasomal degradation, while CSN4 promotes p53 degradation; these effects are mediated through a CSN4-CSN5-CK2 complex, with CK2 regulating stability of both sGCα1 and p53. |
Co-immunoprecipitation; siRNA knockdown of CSN4/CSN5; Western blot for protein stability; CK2 inhibition |
Molecular endocrinology (Baltimore, Md.) |
Medium |
24725084
|
| 2014 |
CSN4 was identified as a Ca2+-dependent binding partner of the EF-hand protein tescalcin; this interaction involves the PCI domain of CSN4; tescalcin upregulation during differentiation coincides with reduced CSN deneddylation of Cul1 and stabilization of p27Kip1, and tescalcin knockdown increases Cul1 deneddylation and Skp2/c-Jun expression while decreasing p27/p53. |
Co-immunoprecipitation; domain mapping; siRNA knockdown; Western blot for deneddylation and downstream targets |
Journal of cell science |
Medium |
24659803
|
| 2014 |
3D cryo-EM structural analysis of human CSN reveals a central horseshoe-shaped segment formed by PCI domain subunits; CSN2 and CSN4 densities are better defined in cryo-EM maps compared to negative stain, contributing to the overall structural model of the complex. |
Negative stain EM and cryo-EM single-particle analysis; in vitro deneddylation assay |
Biochemical and biophysical research communications |
Medium |
24973710
|
| 2014 |
In Drosophila GSCs, CSN4 is specifically required for germline stem cell self-renewal but not differentiation; the differentiation factor Bam sequesters Csn4 from the COP9 complex via protein competition, inactivating COP9's self-renewal function and allowing other Csn proteins to promote differentiation. |
Drosophila genetics; GSC clonal analysis; co-immunoprecipitation of Bam-Csn4; loss-of-function phenotyping |
Nature |
High |
25119050
|
| 2016 |
Structural and kinetic analysis showed that N-terminal domains of Csn4 (and Csn2) play important roles in sensing neddylated CRL substrates and enabling formation of a high-affinity fully active CSN-CRL complex; Csn4 communicates binding of neddylated substrate to Csn5 to activate deneddylase activity, with large conformational changes occurring upon binding. |
Cryo-EM structural analysis; hydrogen-deuterium exchange MS; kinetic binding assays; mutagenesis |
eLife |
High |
27031283
|
| 2019 |
TCTP (Translationally Controlled Tumor Protein) controls G1/S transition by interacting with CSN4; genetic interaction studies in Arabidopsis, tobacco cells, and Drosophila showed that downregulation of CSN4 delays G1/S transition; loss-of-function of TCTP increases the fraction of deneddylated CUL1, suggesting TCTP negatively interferes with COP9 (CSN4) function to maintain CUL1 neddylation. |
Genetic interaction studies; knockdown in multiple organisms; CUL1 neddylation state analysis by immunoblot |
PLoS genetics |
Medium |
30695029
|
| 2019 |
In breast cancer MDA-MB-231 cells, CSN4 knockdown decreases cellular proliferation, increases sub-G1 population (apoptosis), and alters expression of CDK6 and Caspase3, indicating CSN4 modulates cell cycle progression and apoptosis through these regulators. |
Lentivirus-mediated CSN4 knockdown; CCK8 proliferation assay; colony formation; cell cycle analysis; Western blot for CDK6 and Caspase3 |
Yi chuan = Hereditas |
Low |
30992253
|
| 2026 |
CSN4 binds directly to DDB1; loss of CSN4 leads to autophagy-mediated destabilization of DDB1 (the DDB1 K1131R mutant is resistant to CSN4 depletion-induced downregulation), reduced CRL4 assembly, and impaired DNA damage-induced ubiquitination; CSN4 deficiency activates autophagy as an alternative degradation pathway for DDB1. |
Co-immunoprecipitation; siRNA knockdown; site-directed mutagenesis (DDB1 K1131R); CRL4 assembly assay; ubiquitination assay; autophagy pathway analysis |
Cellular and molecular life sciences : CMLS |
Medium |
41686221
|