Amino acid sequence and structural properties of protein p12, an African swine fever virus attachment protein.
The gene encoding the African swine fever virus protein p12, which is involved in virus attachment to the host cell, has been mapped and sequenced in the genome of the Vero-adapted virus strain BA71V. The determination of the N-terminal amino acid sequence and the hybridization of oligonucleotide probes derived from this sequence to cloned restriction fragments allowed the mapping of the gene in fragment EcoRI-O, located in the central region of the viral genome. The DNA sequence of an EcoRI-XbaI fragment showed an open reading frame which is predicted to encode a polypeptide of 61 amino acids. The expression of this open reading frame in rabbit reticulocyte lysates and in Escherichia coli gave rise to a 12-kDa polypeptide that was immunoprecipitated with a monoclonal antibody specific for protein p12. The hydrophilicity profile indicated the existence of a stretch of 22 hydrophobic residues in the central part that may anchor the protein in the virus envelope. Three forms of the protein with apparent molecular masses of 17, 12, and 10 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis have been observed, depending on the presence of 2-mercaptoethanol and alkylation with 4-vinylpyridine, indicating that disulfide bonds are responsible for the multimerization of the protein. This result was in agreement with the existence of a cysteine-rich domain in the C-terminal region of the predicted amino acid sequence. The protein was synthesized at late times of infection, and no posttranslational modifications such as glycosylation, phosphorylation, or fatty acid acylation were detected.
Alcamí
A
A
Centro de Biología Molecular, Facultad de Ciencias, Universidad Autónoma, Madrid, Spain.
Angulo
A
A
López-Otín
C
C
Muñoz
M
M
Freije
J M
JM
Carrascosa
A L
AL
Viñuela
E
E
D000358:African Swine Fever Virus / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002474:Cell-Free System; D002478:Cells, Cultured; D002874:Chromosome Mapping; D004926:Escherichia coli / Q000235:genetics; D005814:Genes, Viral / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D011487:Protein Conformation; D016364:Reading Frames; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000097:blood / Q000235:genetics*
D014764:Viral Proteins; C074550:protein p12, African swine fever virus