A Mutational Analysis of the Transforming Functions of the E8 Protein of Bovine Papillomavirus Type 4

• Published in: Virology (New York, N.Y.) N.Y.), 1999-03, Vol.255 (2), p.385-394
• Main Authors: O'Brien, Vincent, Ashrafi, G.Hossein, Grindlay, G.Joan, Anderson, Ruth, Campo, M.Saveria
• Format: Article
• Language: English
• Subjects: Animals; BOVINAE; Bovine papillomavirus 1 - genetics; Bovine papillomavirus 4; BPV-4; Cattle; Cell Division; Cell Line; cell transformation; Cell Transformation, Viral; cyclin A; Cyclin A - genetics; Gene Expression; Humans; Mutagenesis; Oncogene Proteins, Viral - genetics; Oncogene Proteins, Viral - metabolism; PAPOVAVIRIDAE; Promoter Regions, Genetic; PROTEINAS; PROTEINE; PROTEINS; Serum Albumin, Bovine; Transcriptional Activation
• ISSN: 0042-6822; 1096-0341
• DOI: 10.1006/viro.1998.9563

The E8 protein of BPV-4 contributes to transformation of primary bovine cells (PalFs) by inducing anchorage-independent growth and by down-regulating gap junction intercellular communication, likely due to its binding to 16K ductin. We show here that, in addition, E8 confers on PalF cells the ability to grow in low serum and to escape from contact inhibition (focus formation). E8 also transactivates an exogenous human cyclin A gene promoter, suggesting that overexpression of cyclin A is responsible for the transformed phenotype. Mutant forms of E8 were generated to establish whether the transforming functions of the protein could be segregated. Mutations were introduced both in the hydrophobic domain and in the hydrophilic C-terminal “tail”, and chimeras with BPV-1 E5 were constructed. Cells expressing either wild-type E8 or mutant forms were analyzed for their ability to grow in low serum and in suspension and to form foci. Wild-type E8 and its mutants were also analyzed for their ability to transactivate the cyclin A promoter. We show here that the transforming functions of E8 can be segregated and that both the hydrophilic C-terminal tail and the residue at position 17 in the hydrophobic domain are crucial for E8 functions and for the transactivation of the cyclin A promoter. These results support the hypothesis that the different aspects of cellular transformation brought about by E8 might be due to interaction with different cellular targets. They suggest that E8 might function differently from BPV-1 E5 and demonstrate that the separate domains of E5 and E8 are not functionally interchangeable.