Identification of an epitope on the P and V proteins of simian virus 5 that distinguishes between two isolates with different biological characteristics.
Two canine isolates of simian virus 5 (SV5), termed CPI+ and CPI-, were examined for their ability to react with a bank of monoclonal antibodies (MAbs) that had been previously raised against a human isolate of SV5. CPI- virus was originally isolated from the brain of a gnotobiotic dog infected with CPI+ virus and establishes persistent infections more readily than CPI+ in vitro. Of more than 50 MAbs tested, only one (P-k) reacted with CPI+ but not CPI-, enabling distinction between the two canine isolates. It had been shown previously that MAb P-k reacts with an epitope common to both the P and V proteins. In order to characterize further the epitope binding site of this MAb the P/V genes of CPI+ and CPI- were sequenced. There were four nucleotide differences between CPI+ and CPI-, three of which resulted in predicted amino acid substitutions. Synthetic peptides corresponding to regions encompassing these changes were made and radioimmune competition assays were used to identify the epitope binding site of MAb P-k. Sequence comparison of the P/V gene of CPI+ with the published sequence of a monkey isolate of SV5 (W3) revealed 14 nucleotide differences with five amino acid substitutions. The only amino acid substitution observed between CPI+, CPI- and W3 which altered the predicted secondary structures of the P and V proteins was a leucine to proline change that induced a predicted beta-turn and resulted in the loss of binding of MAb P-k.
Southern
J A
JA
Department of Biochemistry and Microbiology, University of St Andrews, Fife, U.K.
Young
D F
DF
Heaney
F
F
Baumgärtner
W K
WK
Randall
R E
RE
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology*; D001345:Autoradiography; D001483:Base Sequence; D001667:Binding, Competitive; D015153:Blotting, Western; D004279:DNA, Viral / Q000737:chemistry; D004285:Dogs; D000939:Epitopes / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D010750:Phosphoproteins / Q000235:genetics / Q000276:immunology*; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000737:chemistry; D015531:Radioimmunoprecipitation Assay; D018108:Respirovirus / Q000235:genetics / Q000276:immunology*; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D004279:DNA, Viral; D000939:Epitopes; D009841:Oligonucleotides; D010455:Peptides; D010750:Phosphoproteins; D012367:RNA, Viral; D014764:Viral Proteins