title: 44-amino-acid E5 transforming protein of bovine papillomavirus requires a hydrophobic core and specific carboxyl-terminal amino acids.
abstract: The 44-amino-acid E5 protein of bovine papillomavirus type 1 is the shortest known protein with transforming activity. To identify the specific amino acids required for in vitro focus formation in mouse C127 cells, we used oligonucleotide-directed saturation mutagenesis to construct an extensive collection of mutants with missense mutations in the E5 gene. Characterization of mutants with amino acid substitutions in the hydrophobic middle third of the E5 protein indicated that efficient transformation requires a stretch of hydrophobic amino acids but not a specific amino acid sequence in this portion of the protein. Many amino acids in the carboxyl-terminal third of the protein can also undergo substitution without impairment of focus-forming activity, but the amino acids at seven positions, including two cysteine residues that mediate dimer formation, appear essential for efficient transforming activity. These essential amino acids are the most well conserved among related fibropapillomaviruses. The small size of the E5 protein, its lack of similarity to other transforming proteins, and its ability to tolerate many amino acid substitutions implies that it transforms cells via a novel mechanism.
authors.affiliation: Department of Human Genetics, Yale University School of Medicine, New Haven, Connecticut 06510.
authors.forename: B H
authors.initials: BH
authors.lastname: Horwitz
authors.forename: A L
authors.initials: AL
authors.lastname: Burkhardt
authors.forename: R
authors.initials: R
authors.lastname: Schlegel
authors.forename: D
authors.initials: D
authors.lastname: DiMaio
mesh_terms: D000595:Amino Acid Sequence; D052688:Bovine papillomavirus 1 / Q000235:genetics / Q000502:physiology*; D002472:Cell Transformation, Viral; D002874:Chromosome Mapping; D004252:DNA Mutational Analysis; D008969:Molecular Sequence Data; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D027383:Papillomaviridae / Q000502:physiology*; D012995:Solubility; D013329:Structure-Activity Relationship
chemical_list: D009856:Oncogene Proteins, Viral