Basic Information
Remarks
Chemical Identifiers
Basic Properties
Lipinski RO5
Hydrophobic Tag Properties
Mechanism of Action
POI (Protein of Interest)
Amino Acid Sequence
3D Structure (/structures/AF-Q14191-F1-model_v6.cif)
Function
Multifunctional enzyme that has magnesium and ATP-dependent 3'-5' DNA-helicase activity on partially duplex substrates . Also has 3'->5' exonuclease activity towards double-stranded (ds)DNA with a 5'-overhang. Has no nuclease activity towards single-stranded (ss)DNA or blunt-ended dsDNA. Helicase activity is most efficient with (d)ATP, but (d)CTP will substitute with reduced efficiency; strand displacement is enhanced by single-strand binding-protein (heterotrimeric replication protein A complex, RPA1, RPA2, RPA3). Binds preferentially to DNA substrates containing alternate secondary structures, such as replication forks and Holliday junctions. May play an important role in the dissociation of joint DNA molecules that can arise as products of homologous recombination, at stalled replication forks or during DNA repair. Alleviates stalling of DNA polymerases at the site of DNA lesions. Plays a role in the formation of DNA replication focal centers; stably associates with foci elements generating binding sites for RP-A (By similarity). Plays a role in double-strand break repair after gamma-irradiation. Unwinds some G-quadruplex DNA (d(CGG)n tracts); unwinding seems to occur in both 5'-3' and 3'-5' direction and requires a short single-stranded tail. d(CGG)n tracts have a propensity to assemble into tetraplex structures; other G-rich substrates from a telomeric or IgG switch sequence are not unwound. Depletion leads to chromosomal breaks and genome instability.
Degradation Pathway
Mechanism
The ubiquitin-proteasome pathway (UPP) constitutes the principal pathway for protein degradation in eukaryotic cells, mediating the breakdown of target proteins through ubiquitin tagging and subsequent proteasomal degradation. In an ATP-dependent manner, the ubiquitin-activating enzyme (E1) activates ubiquitin (Ub) and conjugates it to a ubiquitin-conjugating enzyme (E2). E3 ligase subsequently catalyzes the transfer of Ub to the target protein, leading to the formation of a polyubiquitin chain that completes the ubiquitination mark. The 19S regulatory particle of the 26S proteasome recognizes this ubiquitination signal and, after deubiquitination, mediates the ATP-dependent delivery of the substrate into the 20S proteasome for degradation.
Description
To elucidate the mechanism of 14c-induced WRN degradation, we first treated HCT-116 cells with the protein synthesis inhibitor cyclo heximide (CHX) in combination with 14c. As shown in Figs. 8A and 14c remarkably accelerated WRN degradation with significant reduction in half-life of WRN. Pretreatment with proteasome inhibitors MG132 (Fig. 8B) or bortezomib (Fig. 8C) completely abrogated 14c-induced WRN depletion, suggesting the involvement of proteasome pathway. In addition, HSP90 inhibitor 17-AAG potentiated 14c-mediated WRN degradation (Fig. 8D), further implicating the ubiquitin-proteasome pathway [34]. This finding was further supported by a co-immunoprecipitation (Co-IP) assay that revealed an increase in WRN ubiquitination following 14c treatment (Fig. 8E). Collectively, these data demonstrate that 14c induces WRN degradation through ubiquitin-proteasome system-mediated proteolysis.
Activity Data
Description
As expected, unlike clinical-stage WRN inhibitors showing MSI-dependent degradation, both 14c and 14d exhibited potent and phenotype-agnostic WRN degradation in a dose-dependent manner across all tested MSI-H and MSS cells (Table 3and Fig. 4). Strikingly, 14c stood out as the most potent degrader in HCT-116, SW-48 and SW-480 cells with DC50 values of 1.7, 3 and 5.9 μM, respectively, as well as excellent Dmax values of >97 %.
Description
As expected, unlike clinical-stage WRN inhibitors showing MSI-dependent degradation, both 14c and 14d exhibited potent and phenotype-agnostic WRN degradation in a dose-dependent manner across all tested MSI-H and MSS cells (Table 3and Fig. 4). Strikingly, 14c stood out as the most potent degrader in HCT-116, SW-48 and SW-480 cells with DC50 values of 1.7, 3 and 5.9 μM, respectively, as well as excellent Dmax values of >97 %.
Description
As expected, unlike clinical-stage WRN inhibitors showing MSI-dependent degradation, both 14c and 14d exhibited potent and phenotype-agnostic WRN degradation in a dose-dependent manner across all tested MSI-H and MSS cells (Table 3and Fig. 4). Strikingly, 14c stood out as the most potent degrader in HCT-116, SW-48 and SW-480 cells with DC50 values of 1.7, 3 and 5.9 μM, respectively, as well as excellent Dmax values of >97 %.
Description
As expected, unlike clinical-stage WRN inhibitors showing MSI-dependent degradation, both 14c and 14d exhibited potent and phenotype-agnostic WRN degradation in a dose-dependent manner across all tested MSI-H and MSS cells (Table 3and Fig. 4). Strikingly, 14c stood out as the most potent degrader in HCT-116, SW-48 and SW-480 cells with DC50 values of 1.7, 3 and 5.9 μM, respectively, as well as excellent Dmax values of >97 %.
Description
As expected, unlike clinical-stage WRN inhibitors showing MSI-dependent degradation, both 14c and 14d exhibited potent and phenotype-agnostic WRN degradation in a dose-dependent manner across all tested MSI-H and MSS cells (Table 3and Fig. 4). Strikingly, 14c stood out as the most potent degrader in HCT-116, SW-48 and SW-480 cells with DC50 values of 1.7, 3 and 5.9 μM, respectively, as well as excellent Dmax values of >97 %.
Description
As expected, unlike clinical-stage WRN inhibitors showing MSI-dependent degradation, both 14c and 14d exhibited potent and phenotype-agnostic WRN degradation in a dose-dependent manner across all tested MSI-H and MSS cells (Table 3and Fig. 4). Strikingly, 14c stood out as the most potent degrader in HCT-116, SW-48 and SW-480 cells with DC50 values of 1.7, 3 and 5.9 μM, respectively, as well as excellent Dmax values of >97 %.
Description
Notably, the enhanced degradative activity of 14c correlated with superior antiproliferative activity compared to 14d and parental inhibitor H3B-219 against MSI-H cells (HCT-116, SW-48). In addition, consistent with its synthetic lethality mechanism, 14c showed negligible cytotoxicity in MSS SW-480 cells (IC50 >100 μM) (Table 3and Fig. 4). Therefore, based on these degradation and antiproliferative data, LdCHT degrader 14c was selected for further activity and mechanism investigations.
Description
Notably, the enhanced degradative activity of 14c correlated with superior antiproliferative activity compared to 14d and parental inhibitor H3B-219 against MSI-H cells (HCT-116, SW-48). In addition, consistent with its synthetic lethality mechanism, 14c showed negligible cytotoxicity in MSS SW-480 cells (IC50 >100 μM) (Table 3and Fig. 4). Therefore, based on these degradation and antiproliferative data, LdCHT degrader 14c was selected for further activity and mechanism investigations.
Description
Notably, the enhanced degradative activity of 14c correlated with superior antiproliferative activity compared to 14d and parental inhibitor H3B-219 against MSI-H cells (HCT-116, SW-48). In addition, consistent with its synthetic lethality mechanism, 14c showed negligible cytotoxicity in MSS SW-480 cells (IC50 >100 μM) (Table 3and Fig. 4). Therefore, based on these degradation and antiproliferative data, LdCHT degrader 14c was selected for further activity and mechanism investigations.
Disease Information
| Disease | ICD-11 | Source |
|---|---|---|
| Colon carcinoma | 2B90.Y | Cell Line Origin |
| Werner syndrome | LD2B | Gene-Disease Association |
| Bloom Syndrome | 4A01.31 | Gene-Disease Association |
| Cataract | 9B10 | Gene-Disease Association |
| Cockayne Syndrome | LD2B | Gene-Disease Association |
| Sarcoma | 2B5F.3 | Gene-Disease Association |
| Endocrine Gland Cancer | 2D1Z | Gene-Disease Association |
| Maturity-Onset Diabetes Of The Young | 5A13.6 | Gene-Disease Association |