E6 proteins from high-risk HPV, low-risk HPV, and animal papillomaviruses activate the Wnt/β-catenin pathway through E6AP-dependent degradation of NHERF1

Abstract

High-risk human papillomavirus (HPV) E6 proteins associate with the cellular ubiquitin ligase E6-Associated Protein (E6AP), and then recruit both p53 and certain cellular PDZ proteins for ubiquitination and degradation by the proteasome. Low-risk HPV E6 proteins also associate with E6AP, yet fail to recruit p53 or PDZ proteins; their E6AP-dependent targets have so far been uncharacterized. We found a cellular PDZ protein called Na+/H+ Exchanger Regulatory Factor 1 (NHERF1) is targeted for degradation by both high and low-risk HPV E6 proteins as well as E6 proteins from diverse non-primate mammalian species. NHERF1 was degraded by E6 in a manner dependent upon E6AP ubiquitin ligase activity but independent of PDZ interactions. A novel structural domain of E6, independent of the p53 recognition domain, was necessary to associate with and degrade NHERF1, and the NHERF1 EB domain was required for E6-mediated degradation. Degradation of NHERF1 by E6 activated canonical Wnt/{beta}-catenin signaling, a key pathway that regulates cell growth and proliferation. Expression levels of NHERF1 increased with increasing cell confluency. This is the first study in which a cellular protein has been identified that is targeted for degradation by both high and low-risk HPV E6 as well as E6 proteins from diverse animal papillomaviruses. This suggests that NHERF1 plays a role in regulating squamous epithelial growth and further suggests that the interaction of E6 proteins with NHERF1 could be a common therapeutic target for multiple papillomavirus types.\n\nAuthor summaryPapillomaviruses cause benign squamous epithelial tumors through the action of virally encoded oncoproteins termed E6 and E7, which are classified as either high or low-risk based upon the propensity of the tumor to evolve into cancer. E6 proteins from both high and low-risk HPVs interact with a cellular ubiquitin ligase called E6AP. High-risk E6 proteins hijack E6AP ubiquitin ligase activity to target p53 for degradation. Degradation targets of the low-risk E6 proteins in complex with E6AP have not been described. Here, we describe a protein called NHERF1 that is targeted for degradation by both high and low-risk E6 proteins, as well as E6 proteins from diverse animal species. Degradation of NHERF1 resulted in activation of an oncogenic cellular signaling pathway called Wnt. Identification of NHERF1 as a highly conserved E6 degradation target could inform therapies directed against both low-risk HPVs and cancer-inducing high-risk HPVs.