Genetic and biochemical definition of the bovine papillomavirus E5 transforming protein.
Mutations surrounding the first methionine codon of the E5 transforming gene of bovine papillomavirus (type 1) were analyzed for their effect on cellular transformation and on the synthesis of the 7-kd E5 polypeptide. Frameshift mutations upstream of this methionine codon (bp 3879) affect neither transforming activity nor the ability to synthesize full-size E5 protein. In contrast, frameshift mutations distal to this position result in the inhibition of cell transformation and prevent synthesis or accumulation of E5 protein in cells containing the mutant viral genomes. Several in-frame mutations distal to the first methionine codon have a minimal effect on transforming activity but alter the electrophoretic mobility of the E5 protein in a manner consistent with the generated genetic alteration (deletion, insertion or substitution). In all cases where the protein is detected, it fractionates with cellular membranes and forms dimers. These studies indicate that (i) the methionine codon at bp 3879 serves as the initiation codon for the mature E5 protein, (ii) changing the charge of the E5 amino-terminus (from neutral to positive) does not prevent the association of this hydrophobic polypeptide with cellular membranes, and (iii) E5 amino-terminal mutations do not interfere with the ability of this polypeptide to form homodimers. We conclude that the major focus-inducing activity of the intact BPV genome is due to the function of the small polypeptide encoded in the 3' half of the E5 ORF.
Burkhardt
A
A
Laboratory of Tumor Virus Biology, National Cancer Institute, Bethesda, MD 20892.
DiMaio
D
D
Schlegel
R
R
D000595:Amino Acid Sequence; D052688:Bovine papillomavirus 1 / Q000235:genetics*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D005796:Genes; D005814:Genes, Viral; D005822:Genetic Vectors; D009154:Mutation; D009856:Oncogene Proteins, Viral / Q000235:genetics* / Q000378:metabolism; D027383:Papillomaviridae / Q000235:genetics*; D010957:Plasmids
D009856:Oncogene Proteins, Viral