MmuPV1 E7’s interaction with PTPN14 delays Epithelial differentiation and contributes to virus-induced skin disease

Human papillomaviruses (HPVs) contribute to approximately 5% of all human cancers. Species-specific barriers limit the ability to study HPV pathogenesis in animal models. Murine papillomavirus (MmuPV1) provides a powerful tool to study the roles of papillomavirus genes in pathogenesis arising from a natural infection. We previously identified Protein Tyrosine Phosphatase Non-Receptor Type 14 (PTPN14), a tumor suppressor targeted by HPV E7 proteins, as a putative cellular target of MmuPV1 E7. Here, we confirmed the MmuPV1 E7-PTPN14 interaction. Based on the published structure of the HPV18 E7/PTPN14 complex, we generated a MmuPV1 E7 mutant, E7(K81S), that was defective for binding PTPN14. Wild-type (WT) and E7(K81S) mutant viral genomes replicated as extrachromosomal circular DNAs to comparable levels in mouse keratinocytes. E7(K81S) mutant virus (E7(K81S) MmuPV1) was generated and used to infect FoxN/Nude mice. E7(K81S) MmuPV1 caused neoplastic lesions at a frequency similar to that of WT MmuPV1, but the lesions arose later and were smaller than WT-induced lesions. The E7(K81S) MmuPV1-induced lesions also had a trend towards a less severe grade of neoplastic disease. In the lesions, E7(K81S) MmuPV1 supported the late (productive) stage of the viral life cycle and promoted E2F activity and cellular DNA synthesis in suprabasal epithelial cells to similar degrees as WT MmuPV1. There was a similar frequency of lateral spread of infections among mice infected with E7(K81S) or WT MmuPV1. Compared to WT MmuPV1-induced lesions, E7(K81S) MmuPV1-induced lesions had a significant expansion of cells expressing differentiation markers, Keratin 10 and Involucrin. We conclude that an intact PTPN14 binding site is necessary for MmuPV1 E7’s ability to contribute to papillomavirus-induced pathogenesis and this correlates with MmuPV1 E7 causing a delay in epithelial differentiation, which is a hallmark of papillomavirus-induced neoplasia.