| 2019 |
Crystal structure of the DCAF15-DDB1-DDA1-indisulam-RBM39(RRM2) complex at 2.3 Å resolution revealed that DCAF15 has a distinct topology that embraces the RBM39 RRM2 domain largely via non-polar interactions, with indisulam binding in a pocket between DCAF15 and RBM39(RRM2) to coordinate additional protein-protein contacts. Point mutants of RBM39 and indisulam analogs validated the structural model and defined the RBM39 α-helical degron motif recognized by DCAF15. |
X-ray crystallography (2.3 Å), RBM39 point mutagenesis, indisulam analog structure-activity studies |
Nature chemical biology |
High |
31819272
|
| 2019 |
Cryo-EM structure of the DDB1-DCAF15-DDA1 core ligase complex bound to RBM39 and E7820 at 4.4 Å, together with crystal structures of engineered subcomplexes, showed that DCAF15 adopts a new fold stabilized by DDA1 and that aryl-sulfonamides neo-functionalize a shallow, non-conserved pocket on DCAF15 to selectively recruit and degrade RBM39 and RBM23. Extensive protein-protein contacts between the ligase and substrate mitigate low-affinity interactions between the drug and DCAF15. |
Cryo-EM (4.4 Å), X-ray crystallography of subcomplexes, biochemical binding assays |
Nature chemical biology |
High |
31686031
|
| 2019 |
Crystal structure of DDA1-DDB1-DCAF15 in complex with E7820 and the RRM2 domain of RBM39 revealed that E7820 packs in a shallow pocket on the surface of DCAF15 and the modified interface binds RBM39 through the α1 helix of the RRM2 domain. Kinetic studies demonstrated that aryl sulfonamide and RBM39 bind DCAF15 in a synergistic (cooperative) manner, confirming aryl sulfonamides act as molecular glues. |
X-ray crystallography, kinetic binding analysis |
Structure |
High |
31693911
|
| 2019 |
Domain mapping and random mutagenesis identified DCAF15 residues Q232 and D475 as necessary for indisulam-dependent RBM39 recruitment. RBM39 is recruited to DCAF15 via its RRM2 domain and is ubiquitinated on its N terminus. RBM23, a paralog of RBM39, is also recruited to CRL4-DCAF15 through its RRM2 domain and undergoes sulfonamide-dependent degradation. Indisulam-induced splicing changes (intron retention, exon skipping) are attributable solely to RBM39 degradation. |
Domain mapping, random mutagenesis, ubiquitination assays, RNA-seq |
Cell reports |
High |
31693891
|
| 2019 |
Using pSILAC proteomics combined with two complementary digestion approaches, PRPF39 was identified as a novel substrate of E7070/DCAF15. PRPF39 undergoes DCAF15-dependent ubiquitination and proteasomal degradation upon E7070 treatment, establishing it as a bona fide neo-substrate of the CUL4-DCAF15 E3 ubiquitin ligase. |
pSILAC mass spectrometry, ubiquitination assay, proteasome inhibitor rescue, DCAF15 knockdown |
Journal of proteomics |
Medium |
31626998
|
| 2020 |
ARNT (aryl hydrocarbon receptor nuclear translocator) interacts with DCAF15 and undergoes proteasomal degradation induced by aryl sulfonamides indisulam and E7820 through CRL4-DCAF15, independently of the known neo-substrates RBM39 and RBM23. This degradation inhibited transcriptional activities of HIFs and AhR associated with ARNT. |
Co-immunoprecipitation, proteasome inhibitor rescue, RBM39/RBM23 knockdown controls, transcriptional reporter assays |
Molecules and cells |
Medium |
33168788
|
| 2021 |
ZEB1 was identified as an endogenous substrate of the CRL4-DCAF15 E3 ubiquitin ligase complex. DCAF15 acts as an adaptor that specifically recognizes the N-terminal zinc finger domain of ZEB1 and triggers its ubiquitin-proteasome-mediated degradation. DCAF15 knockdown led to ZEB1 upregulation and EMT activation, while DCAF15 overexpression suppressed ZEB1 and inhibited EMT in hepatocellular carcinoma cells. |
Co-immunoprecipitation, domain mapping, ubiquitination assay, DCAF15 knockdown/overexpression with EMT phenotypic readout |
Aging |
Medium |
33833131
|
| 2021 |
DCAF15/RBM39 pathway controls KRAS4A alternative splicing. Pharmacological inhibition of RBM39 using indisulam reduces KRAS4A isoform levels and inhibits cancer stem cells, placing DCAF15 upstream of KRAS4A isoform production through RBM39-dependent splicing regulation. |
Indisulam treatment, isoform-specific RT-PCR/RNA analysis, cancer stem cell functional assays |
Nature communications |
Medium |
34257283
|
| 2024 |
DCAF15 directly interacts with the SMC1A subunit of the cohesin complex and destabilizes cohesin regulatory factors PDS5A and CDCA5. Loss of DCAF15 causes loss of PDS5A and CDCA5, precluding cohesin acetylation on chromatin, resulting in uncontrolled chromatin loop extrusion, defective DNA replication fork integrity, accumulation of DNA damage, and apoptosis in acute myeloid leukemia (AML) cells. |
Domain-focused genetic screen, DCAF15 loss-of-function (knockout), co-immunoprecipitation (DCAF15-SMC1A), cohesin acetylation assays, chromatin loop extrusion analysis, replication fork assays, DNA damage markers |
Nature communications |
High |
38961054
|
| 2025 |
DCAF15 interacts with EphA2 receptor and promotes its K48- and K63-linked ubiquitination at lysine residues K646, K649, and K754, leading to EphA2 degradation. S. suis STK phosphorylates SIRT1 at S48, triggering SIRT1-mediated deacetylation of DCAF15 at K552 and K581, which enhances DCAF15-EphA2 interaction and EphA2 ubiquitination. This DCAF15-mediated EphA2 degradation disrupts blood-brain barrier integrity during S. suis meningitis. |
Co-immunoprecipitation, ubiquitination site mapping (K646/K649/K754), acetylation site mapping (K552/K581), SIRT1 deacetylase assay, in vitro and in vivo BBB disruption models |
Communications biology |
Medium |
41276610
|
| 2024 |
Network-based proteomics analysis identified RBM5 as a novel indisulam neo-substrate degraded in a proteasome-dependent manner via DCAF15, expanding the list of CRL4-DCAF15 substrates beyond RBM39 and RBM23. |
Network-based omics prioritization (Random Walk with Restart), proteomics, proteasome inhibitor rescue |
bioRxivpreprint |
Low |
bio_10.1101_2024.09.16.613231
|