Loss of the proteasome deubiquitinase USP14 induces growth defects and a senescence phenotype in colorectal cancer cells.
The proteasome-associated deubiquitinase USP14 has attracted significant attention in recent years due to its potential as a drug target. Here, we employ an inducible USP14 knockout system to dissect early phenotypic and gene expression alterations following USP14 loss. Transcriptomic analysis revealed widespread changes in the expression of actin cytoskeleton components associated with shifts in cellular actin distribution and morphology, as well as changes in the deubiquitinase expression profile. We observed an increase in ubiquitin turnover that appeared to be offset by the upregulation of polyubiquitin genes UBB and UBC. Overall design: HCT116 cells bearing floxed USP14 exon 5 was lentivirally transduced with either Cre recombinase + GFP or GFP alone. Cre transduction results in the complete knockout of USP14. Cre + GFP - and GFP-transduced cells were enriched through flow cytometry and expanded. At D14 post transduction, the cells were harvested for bulk RNA sequencing