Opposing effects of bovine papillomavirus type 1 E6 and E7 genes on Fas-mediated apoptosis

Abstract

Programmed cell death (PCD), best exemplified by apoptosis, is a genetically programmed process of cellular destruction that is indispensable for normal development and homeostasis of multicellular organisms. Tumor necrosis factor alpha (TNF) and related cytokines are employed by host defenses to eliminate virally infected cells through induction of apoptosis. Many viruses have evolved specific gene products to modulate this process. We have recently shown that the bovine papillomavirus type 1 (BPV-1) E6 and E7 genes independently sensitize mouse cells to TNF-induced apoptosis. In this report, we investigated the effect of E6 and E7 expression on Fas-mediated apoptosis. In contrast to TNF-mediated apoptosis, E6 and E7 demonstrated opposite effects: while E7 potentiated apoptosis triggered by an agonistic Fas antibody, E6 attenuated the effect. The mitochondrial pathway leading to the activation of caspases appears to be involved in Fas-mediated apoptosis in C127 cells. To further explore the mechanisms by which E6 and E7 modulate Fas-mediated apoptosis, we examined the surface expression of Fas in cells expressing E6 and E7. Significantly, levels of surface Fas expression correlated with the opposing effects of E6 and E7 on Fas-mediated apoptosis. Specifically, while E7 increased the surface expression of Fas, E6 reduced surface Fas expression. Mutational analysis demonstrated a correlation of E6's ability to downregulate surface Fas expression and apoptosis. Since the tumor suppressor p53 can be targeted for degradation by human papillomavirus and has been shown to induce apoptosis by upregulating surface Fas expression, we investigated the role of p53 in BPV-1 E6 and E7 modulation of Fas-mediated apoptosis. Our results demonstrated that the modulatory effects by E6 and E7 could occur in the absence of p53. Interestingly, the reduced Fas protein level on the cell surface is not accompanied by a decrease in total Fas levels in E6-expressing cells. Instead, considerably more Fas protein is found in the cytoplasm of cells expressing E6. These results highlight a novel activity of E6 and E7 that may be involved in viral pathogenesis.