Mutations in the human papillomavirus type 16 E2 protein identify a region of the protein involved in binding to E1 protein.
Papillomavirus DNA replication is primarily dependent upon two viral gene products, E1 and E2. Work with bovine papillomavirus has shown that the E2 protein can bind directly to the E1 protein and enhance the binding of E1 to the viral origin of replication. However, little is known about the mechanism of interaction between E1 and E2 proteins. In this study we have analysed in detail the association between human papillomavirus type 16 (HPV-16) E1 and E2 proteins. Using a purified glutathione S-transferase-HPV-16 E1 fusion protein from Escherichia coli and E2 proteins produced by in vitro transcription-translation, we have developed a rapid and simple method for investigating the association between E1 and E2 in vitro. The binding of E2 to E1 was found to be dependent on sequences in the N-terminal activation domain of the E2 protein. Truncated forms of E2, including a putative repressor form of E2 encoding the DNA binding domain, failed to associate with E1 in this assay. The region of E2 required for efficient binding to E1 was then localized using mutants in the activation domain of E2. These results demonstrated that only a short region of E2 was required for association with E1. This region of E2 was found to be highly conserved amongst all papillomaviruses, suggesting a conservation of E2 function and a common mechanism of interaction between these virally encoded proteins.
Storey
A
A
International Centre for Genetic Engineering and Biotechnology, Trieste, Italy. a.storey@lhmc.ac.uk
Piccini
A
A
Massimi
P
P
Bouvard
V
V
Banks
L
L
D001665:Binding Sites; D004268:DNA-Binding Proteins; D005982:Glutathione Transferase / Q000235:genetics; D006801:Humans; D009838:Oligodeoxyribonucleotides / Q000378:metabolism; D015513:Oncogene Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D009856:Oncogene Proteins, Viral / Q000378:metabolism*; D027383:Papillomaviridae / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
D004268:DNA-Binding Proteins; C093478:E1 protein, Human papillomavirus 16; C054855:E2 protein, Human papillomavirus type 16; D009838:Oligodeoxyribonucleotides; D015513:Oncogene Proteins; D009856:Oncogene Proteins, Viral; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase