Updated project metadata. The goal of the present project was to investigate the EGF-dependent ubiquitination changes occurring in HeLa cells upon a CYLD deficiency. We applied SILAC-based quantitative mass spectrometry proteomics together with genetic engineering and biochemical approaches, in order to enrich the ubiquitinated proteins of the control and silenced cells in response to the growth factor. In order to get a deeper insight of the obtained results, we subsequently combined the enrichment of CYLD protein through immunoprecipitation with SILAC-based proteomics. This allowed us to decipher a specific tyrosine phosphorylation of CYLD that responds to the treatment with EGF and, therefore, we next aimed to analyse the ability of this modification to recruit potential interaction partners. We thus performed a peptide pull-down assay that uncovered a set of proteins specifically associated to the phosphorylated tyrosine and helped to uncover an unexpected, previously unreported role for this deubiquitinase.