The Usp8 deubiquitination enzyme is post-translationally modified by tyrosine and serine phosphorylation

► Src enhances constitutive and ligand-induced Usp8 tyrosine phosphorylation. ► TGFα-induced EGFR endosomal recycling leads to lower Usp8 tyrosine phosphorylation. ► The Usp8 MIT-domain is required for EGF-induced Usp8 1-504 tyrosine phosphorylation. ► The Usp8 Y679F mutation does not abolish phosphoserine-dependent 14-3-3 binding. ► Usp8 is a target for the post-translational serine and tyrosine phosphorylation.