Mapping and nucleotide sequence of the vaccinia virus gene that encodes a 14-kilodalton fusion protein.
A library of rabbit poxvirus DNA fragments contained in the expression cloning vector lambda gt11 was screened with monoclonal antibodies that react specifically against a 14-kilodalton envelope protein of vaccinia virus and rabbit poxvirus. The 14-kilodalton protein appears to play an important role in virus penetration at the level of cell fusion; it also elicits neutralizing antibodies, and it forms covalently linked trimers on the surface of virions and in infected cells (Rodriguez et al., J. Virol. 56:482-488, 1985; Rodriguez et al., J. Virol. 61:395-404, 1987). Two recombinant bacteriophages expressing beta-galactosidase fusion proteins were isolated. Restriction enzyme analysis and hybridization studies mapped the 14-kilodalton encoding sequences in the middle of vaccinia virus HindIII A DNA fragment. Nucleotide sequence analysis revealed an open reading frame (ATG) preceded by a characteristic TAA sequence of late genes. The sequence spans 330 nucleotides and codes for a protein with a molecular weight of 12,500 and an isoelectric point of 6.3. There are two small hydrophobic regions, one at the C terminus (11 amino acids) and the other at the N terminus (5 amino acids). The protein contains two cysteines for oligomer formation and one glycosylation site. Inspection of the deduced amino acid sequence of the 14-kilodalton protein revealed consensus sites with the hemagglutinin precursor of influenza A virus and with adenylate kinase and cytochrome c of various species.
Rodriguez
J F
JF
Department of Biochemistry, State University of New York Health Science Center at Brooklyn, New York 11203-2098.
Esteban
M
M
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D004262:DNA Restriction Enzymes; D005796:Genes; D005814:Genes, Viral; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009710:Nucleotide Mapping; D012689:Sequence Homology, Nucleic Acid; D014616:Vaccinia virus / Q000235:genetics*; D014760:Viral Fusion Proteins / Q000235:genetics* / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D014760:Viral Fusion Proteins; D004262:DNA Restriction Enzymes