| 2003 |
Crystal structure of the 120 kDa quaternary complex of human APPBP1-UBA3 (heterodimeric E1), NEDD8, and ATP was solved, revealing a bipartite interface for selective NEDD8 recruitment involving a domain common to all ubl-activating enzymes plus eukaryotic E1-specific sequences. A single conserved arginine in APPBP1-UBA3 acts as a selectivity gate preventing misactivation of ubiquitin by NEDD8's E1. |
X-ray crystallography of quaternary complex + mutational analysis modeling ubiquitin into NEDD8 binding site |
Molecular cell |
High |
14690597
|
| 2003 |
Human APPBP1-UBA3 catalyzes NEDD8 activation via a pseudo-ordered mechanism (ATP leading, NEDD8 trailing substrate), forming a stable ternary complex of NEDD8-adenylate and Uba3-NEDD8 thioester intermediate, and then transferring NEDD8 to HsUbc12 (kcat = 3.5 s⁻¹). Alanine 72 of NEDD8 is a critical specificity determinant for APPBP1-UBA3 binding, since the Ub(R72L) mutant can serve as substrate. Wild-type ubiquitin fails to support activation or HsUbc12 transthiolation. |
In vitro enzyme kinetics with radiolabeled ATP and 125I-NEDD8, isotope exchange, transthiolation assays, mutagenesis |
The Journal of biological chemistry |
High |
12740388
|
| 2003 |
In fission yeast, But1 and But2 proteins physically bind to Uba3 (the catalytic subunit of NEDD8 E1). But1 is a nuclear protein and its overexpression causes cell elongation (a phenotype of NEDD8 pathway defective mutants); overexpression of but1+ in a ned8-ts mutant was deleterious at permissive temperatures, suggesting But1 has an inhibitory role in the NEDD8 pathway. |
Two-hybrid screen, overexpression phenotype analysis, genetic interaction with ned8-ts mutant |
Biochemical and biophysical research communications |
Medium |
14623327
|
| 2012 |
The C-terminal ubiquitin-like β-grasp domain of human Uba3 (Uba3-βGD) is independently folded in solution and its E2-binding surface undergoes conformational exchange between multiple substates (residues absent from NMR spectrum in free form, adopting a kinked α-helix when complexed with E2), analogous to the E2-binding surface of SUMO E1. |
NMR spectroscopy of isolated Uba3 β-grasp domain, comparison with crystal structures |
Proteins |
Medium |
22821745
|
| 2014 |
UBA3 point mutations I310N (in K562 cells) and Y352H (in U937 cells) confer resistance to the NAE inhibitor MLN4924 by increasing the enzyme's affinity for ATP while decreasing its affinity for NEDD8, thereby reducing MLN4924 potency while preserving sufficient NAE activity for cell survival. |
Selection of resistant cell lines by chronic MLN4924 exposure, sequencing, biochemical analyses of mutant enzyme kinetics |
PloS one |
Medium |
24691136
|
| 2018 |
Using quantitative FRET, UBA3 alone (without APPBP1) is sufficient for NEDD8 activation, whereas APPBP1 functions mainly as a scaffold to accelerate the reaction rate. This contrasts with SUMO E1, where both heterodimer subunits are required for activation. |
Quantitative FRET assays with individual subunits, aided by AESOP computational electrostatic analysis |
Scientific reports |
Medium |
29973603
|
| 2023 |
UBE1C (UBA3)-mediated neddylation of p53 inhibits p53 acetylation at K373, suppresses p53 transcriptional activity, and reduces expression of p53 downstream targets p21 and PTEN. Overexpression of UBE1C/UBA3 and NEDD8 promotes lung cancer cell migration, invasion, and proliferation. |
Overexpression of UBE1C and NEDD8 in lung cancer cells, co-IP/Western blot for p53 neddylation and acetylation, promoter activity assays, functional cell assays |
FASEB journal |
Medium |
37668436
|
| 2023 |
UBA3-dependent neddylation in lung adenocarcinoma cells promotes NF-κB signaling by preventing IκBα phosphorylation/accumulation; blocking UBA3 increases p-IκBα and reduces gene expression of tumor-derived cytokines (CCL2, CXCL1, CXCL2, CSF1, CSF2, IL-6, IL-1B), and decreases recruitment of immunosuppressive cells (TAMs, pDCs, Th2, Tregs). |
mRNA sequencing, functional enrichment, Western blotting, real-time PCR after UBA3 knockdown in LUAD cells |
Medical oncology |
Low |
37656220
|
| 2024 |
UBA3 promotes intrahepatic cholangiocarcinoma proliferation, invasion, and migration by affecting ANXA2 through the MAPK signaling pathway; knockdown of UBA3 inhibits these processes. Bufalin was identified as a targeting inhibitor of UBA3 that suppresses ICC development through this pathway. |
shRNA knockdown of UBA3 in ICC cell lines, mechanistic experiments linking UBA3 to ANXA2/MAPK, bufalin inhibitor studies |
Acta biochimica et biophysica Sinica |
Low |
38298057
|
| 2026 |
NAE1/UBA3 and UBE2M function as E1 and E2 enzymes, respectively, for the urmylation (URM1 conjugation) pathway in human cells under normal and oxidative stress conditions. An activity-based URM1 probe covalently captured cysteine enzymes in the URM1 signaling pathway, identifying this new substrate specificity. Pharmacologic inhibition of NAE1/UBA3 by pevonedistat blocks protein urmylation and exhibits synergy with cisplatin in killing liver cancer cells. |
Activity-based URM1 probe capture, proteomic characterization, cell-based validation, pharmacologic perturbation |
Nature communications |
Medium |
42056084
|