| 1996 |
A single amino acid change in D123 (CDC123) protein causes temperature-sensitive G1-phase arrest in rat fibroblast 3Y1tsD123 cells; the mutant protein is expressed at much lower levels than wild-type, establishing that CDC123 protein quantity is required for cell cycle progression through G1. |
Functional complementation cloning, point mutation identification by RT-PCR/sequencing, western blot quantification of protein levels |
Experimental cell research |
Medium |
8601400
|
| 1999 |
Temperature-sensitive G1 arrest in 3Y1tsD123 cells is caused by increased proteasome-mediated degradation of the mutated D123 protein; overexpression of the mutant protein rescues the cell cycle defect by exceeding the degradation capacity. |
Cycloheximide chase to measure protein stability, cDNA overexpression rescue |
Cell structure and function |
Medium |
10698258
|
| 2001 |
Mutated D123 protein undergoes a proteasome-dependent modification (increased molecular weight, not ubiquitin) prior to degradation; selective proteasome inhibitors lactacystin and MG132 block this degradation and rescue temperature-sensitive growth arrest. |
Proteasome inhibitor treatment (lactacystin, MG132), western blot, somatic cell hybridization |
Cell structure and function |
Medium |
11699637
|
| 2004 |
Yeast Cdc123 (ortholog of mammalian D123/CDC123) physically interacts with Gcd11 (eIF2γ) and controls its abundance; loss of cdc123 depletes Gcd11 (eIF2γ) and causes G1 arrest, placing Cdc123 in an essential pathway for nutritional control of START running parallel to the Tor-Gcn2-Sui2 system. CHF1/CHF2 (RING checkpoint proteins) associate with Cdc123 via the Thr-274 phosphorylatable site and counteract its cell cycle-promoting activity. |
Yeast genetics (cdc123 mutant isolation, suppressor/epistasis analysis), genetic interaction with CHF1/CHF2, protein interaction mapping (binding-site mutagenesis), western blot of Gcd11 levels |
The Journal of biological chemistry |
High |
15319434
|
| 2013 |
Cdc123 functions as a dedicated assembly factor for the eIF2 heterotrimer: it binds unassembled eIF2γ (but not the assembled eIF2 complex) via the C-terminal domain III of eIF2γ; this interaction is necessary and sufficient for eIF2α/β association with eIF2γ. Mutations disrupting Cdc123–eIF2γ binding abolish eIF2 assembly and cause loss of eIF2 activity (reduced polysomes, elevated GCN4 translation). High-level overexpression of all three eIF2 subunits rescues an otherwise lethal cdc123 deletion. |
Yeast CDC123 deletion, polysome profiling, GCN4-lacZ reporter, co-immunoprecipitation of eIF2 subunits, domain mapping with eIF2γ truncations, rescue by subunit overexpression |
The Journal of biological chemistry |
High |
23775072
|
| 2015 |
Crystal structure of S. pombe Cdc123 alone and in complex with domain III of S. cerevisiae eIF2γ shows Cdc123 is an ATP-grasp enzyme; it binds ATP-Mg²⁺ via conserved residues, and mutagenesis of those residues abolishes eIF2 assembly and cell viability, demonstrating that ATP binding is required for Cdc123 function. Domain III of eIF2γ binds domain I of Cdc123. |
X-ray crystallography (structures of apo and eIF2γD3-bound Cdc123), site-directed mutagenesis of ATP-contact residues, yeast viability assays, biochemical binding assays |
Structure |
High |
26211610
|
| 2015 |
Computational and phylogenetic analysis classifies CDC123 as a novel clade (R2K) of ATP-grasp enzymes distinguished by a RAGNYA domain with two conserved lysines; the enzymatic classification predicts CDC123 may function as an ATP-dependent protein-peptide ligase that modifies substrates by oligopeptide tagging. |
Bioinformatic sequence/structure analysis integrated with published biochemical data |
Biology direct |
Low |
25976611
|
| 2023 |
Crystal structure of human CDC123 bound to domain 3 of human eIF2γ shows that eIF2γD3 contacts domain 1 of Cdc123, and the long C-terminal region of human Cdc123 links the ATP-binding site to the eIF2γ-binding site. Thermal shift assay shows ATP binds Cdc123 tightly whereas ADP affinity is much lower. Yeast viability experiments, western blot, and two-hybrid assays confirm that ATP binding is required for human CDC123 function in eIF2 assembly. |
X-ray crystallography of human CDC123–eIF2γD3 complex, thermal shift assay, yeast complementation viability assay, western blot, yeast two-hybrid |
Journal of structural biology |
High |
37507029
|
| 2023 |
In plants, CDC123 is required for ETI (effector-triggered immunity)-associated global translational induction; an increase in cellular ATP concentration during ETI facilitates CDC123-mediated eIF2 complex assembly, linking NLR-dependent ATP elevation to translational reprogramming in defense. |
Genetic screen with translational reporter, CDC123 loss-of-function phenotyping, ATP measurement, eIF2 assembly assays |
Cell host & microbe |
Medium |
36801014
|
| 2025 |
Human CDC123 acts as an ATPase (not merely an ATP-binding scaffold) to drive eIF2 heterotrimer assembly; impaired CDC123 activity reduces eIF2 complex assembly and activates the integrated stress response (ISR) through a noncanonical mechanism, altering global and mRNA-specific translation. Pharmacological or genetic rescue strategies can correct the translational defects caused by impaired CDC123. |
ATPase activity assays, eIF2 complex assembly measurements, translational assays (polysome profiling or reporters), ISR marker readouts, pharmacological and genetic rescue |
The Journal of biological chemistry |
High |
41461316
|
| 2026 |
Cannabidiol (CBD) was identified as an inhibitor of the CDC123–eIF2γ protein-protein interaction; disruption of the CDC123–eIF2γ complex by CBD leads to sustained activation of the integrated stress response and apoptosis in colorectal cancer cells, validating the CDC123–eIF2γ interface as a druggable target. |
Stability- and degradation-based proteome profiling (SDPP) for target ID, biochemical PPI inhibition assays, ISR activation assays, cell viability/apoptosis assays in CRC cell lines |
Journal of the American Chemical Society |
Medium |
41518300
|
| 2014 |
A type 2 diabetes-associated SNP (rs11257655) in the CDC123/CAMK1D locus shows allele-specific enhancer activity in insulinoma and hepatocellular carcinoma cells; the risk allele T binds FOXA1 and FOXA2 with higher affinity than the non-risk allele C, as shown by EMSA, supershift, and allele-specific ChIP in human islets, suggesting that altered FOXA1/FOXA2 binding at this locus modulates CDC123 transcription. |
Luciferase reporter enhancer assays, EMSA, supershift assays, allele-specific ChIP in human islets |
PLoS genetics |
Medium |
25211022
|