Usp12 stabilizes the T-cell receptor complex at the cell surface during signaling

摘要: Posttranslational modifications are central to the spatial and temporal regulation of protein function. Among others, phosphorylation ubiquitylation known regulate proximal T-cell receptor (TCR) signaling. Here we used a systematic unbiased approach uncover deubiquitylating enzymes (DUBs) that participate during TCR signaling in primary mouse T lymphocytes. Using C-terminally modified vinyl methyl ester variant ubiquitin (HA-Ub-VME), captured DUBs differentially recruited cytosol on activation. We identified ubiquitin-specific peptidase (Usp) 12 Usp46, which had not been previously described this pathway. Stimulation with anti-CD3 resulted time-dependent translocation Usp12 from nucleus cytosol. Usp12−/− Jurkat cells displayed defective NFκB, NFAT, MAPK activities owing attenuated surface expression TCR, were rescued reconstitution wild type Usp12. Proximity-based labeling BirA-Usp12 revealed several adaptor proteins acting as interactors stimulated cells, LAT Trat1 reduced cells. demonstrate deubiquitylates prevents lysosomal degradation maintain complex for duration Our benefits use activity-based probes without any previous genome modification, underscores importance ubiquitin-mediated refine cascades.