USP15 inhibits HPV16 E6 degradation and catalytically inactive USP15 has reduced inhibitory activity.

High-risk human papillomaviruses (HPVs) possess transforming activity leading to development of the cancer, including oropharyngeal, anal, penile, vulvar, vaginal, and cervical cancer. The stability of E6 is essential for its complete function as an oncoprotein. Using the yeast two-hybrid system, we identified ubiquitin-specific protease 15 (USP15) as an HPV16 E6-interacting protein. USP15 cleaves polyubiquitin chains of HPV16 E6 and/or ubiquitin precursors. Our results indicate that USP15 could increase the level of HPV16 E6 by inhibiting E6 degradation. USP15 inhibited the degradation of HPV16 E6 in dose-dependent manner. In contrast, catalytically inactive mutants of USP15 had a reduced inhibitory effect on E6 degradation. In particular, USP15 mutants of all three cysteine boxes and the NHL mutant of the KRF box had a drastically reduced inhibitory effect on HPV16 E6 degradation. In addition, HPV16 E6 mRNA was not induced by USP15; therefore, HPV16 E6 appears to be post-translationally regulated. These results suggest that USP15 has the ability to stabilize E6 as a deubiquitinating enzyme, and as an oncoprotein affects biological functions in infected human cells.