IndraLab

"Immunoprecipitation of ectopic FLAG-TDP1 variants (WT, R361K, or R586K) showed a marked increase in the interaction of endogenous UCHL3 with ectopic R586K mutant TDP1, both in the presence and in the [MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"

"Furthermore, compromised TDP1 proteostasis due to defective TDP1-R586 methylation impairs UCHL3-TDP1 binding and accumulates TDP1 in the cells (model)."

"Additionally, UCLH3 knockdown cells show enrichment of ubiquitylated-TDP1 R586K ( Figure 3 G), whereas overexpression of PRMT5 abrogates the binding of TDP1 to UCHL3 ( Figure 3 H), suggesting that met[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"

"To further test the PRMT5 dependence on the UCHL3-TDP1 binding, we overexpressed FLAG-PRMT5 and co-immunoprecipitated ectopic GFP-TDP1."

"Unlike the methylation mutant TDP1 R586K , we detected reduced binding of UCHL3 with TDP1 WT or TDP1 R361K in the absence of CPT ( Figures 3 D and 3F), which corresponds to the enrichment of ubiquityl[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"

"Here, we uncover the crosstalk between TDP1 arginine methylation at R586 and ubiquitylation, a determining factor for the association of TDP1 with UCHL3, thus acting as a switch for regulation of endo[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"

"Unlike the methylation mutant TDP1 R586K , we detected reduced binding of UCHL3 with TDP1 WT or TDP1 R361K in the absence of CPT ( Figures 3 D and 3F), which corresponds to the enrichment of ubiquityl[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"

"Here, we report multiple evidence showing a direct role of UCHL3 in controlling TDP1 proteostasis: (1) reduction of K48-ubiquitylated TDP1 in cancer cells overexpressing UCHL3, (2) reduction of K48-uibiquitylated TDP1 upon addition of recombinant UCHL3, (3) enrichment of K48-ubiquitylated TDP1 upon MG132 treatment, which is dependent on UCHL3 levels, (4) epistatic relationship showing that co-depletion of TDP1 and UCHL3 does not further sensitize cells to TOP1 poisons than depletion of either enzyme alone, (5) physical interaction between TDP1 and UCHL3, and (6) changes in the ubiquitylation status and TDP1 turnover upon changes in UCHL3 levels induced by its ectopic overexpression, siRNA depletion, CRISPR editing, and pharmacologic inhibition and, also, in two physiologically relevant clinical settings."

"We detected a marked increase in the enrichment of ubiquitylated TDP1 R586K (Ub-TDP1 R586K ) in UCLH3 knockdown cells compared with UCHL3-proficient cells ( Figure 3 G, Ub-TDP1), consistent with methy[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"

"Additionally, UCLH3 knockdown cells show enrichment of ubiquitylated-TDP1 R586K ( Figure 3 G), whereas overexpression of PRMT5 abrogates the binding of TDP1 to UCHL3 ( Figure 3 H), suggesting that met[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"

"Furthermore, incubation of purified K48-ubiquitylated TDP1 with recombinant UCHL3 led to reduction of TDP1 ubiquitylation ( xref B) and a physical interaction between TDP1 and UCHL3 was observed by co-immunoprecipitation ( xref C)."

No evidence text available

"Furthermore, incubation of purified K48 ubiquitylated TDP1 with recombinant UCHL3 led to reduction of TDP1 ubiquitylation (XREF_FIG B) and a physical interaction between TDP1 and UCHL3 was observed by co-immunoprecipitation (XREF_FIG C)."

"Remarkably, the association of TDP1 R586K with UCHL3 remains unchanged even in the presence of DNA damage with CPT ( Figure 3 D, UCHL3, and 3F)."