Prediction and demonstration of a novel Epstein-Barr virus nuclear antigen.
The protein sequence predicted by the Epstein Barr virus (EBV) BERF4 open reading frame includes a tetrapeptide, Lys-Arg-Pro-Arg (KRPR), shown for other proteins to be a component of a signal for rapid nuclear localization. A subgenomic fragment of EBV DNA containing BERF4 has been incorporated into an expression vector, transfected onto primate cells and the nuclear distribution of the resulting protein established by immunofluorescence using EBV positive human sera. These sera contained high titres of antibodies to a fusion protein, produced in E. coli, consisting of beta-galactosidase and the C-terminal 167 amino acids of BERF4. Immunoaffinity purified antibodies reactive with the EBV component of the fusion show the molecular weight of this antigen in EBV immortalized B-cell lines to be about 160 kD. The demonstration that BERF4 contains an exon encoding a nuclear protein identifies a new EBNA gene (EBNA-6) and suggests that KRPR is a signal sequence common to a number of viral and cellular nuclear polypeptides which bind to nucleic acids and may therefore be of predictive value in identifying karyophilic proteins.
Allday
M J
MJ
Department of Virology, Royal Postgraduate Medical School, London, UK.
Crawford
D H
DH
Griffin
B E
BE
D000595:Amino Acid Sequence; D000818:Animals; D000956:Antigens, Viral / Q000235:genetics*; D002460:Cell Line; D002467:Cell Nucleus / Q000276:immunology; D003001:Cloning, Molecular; D019309:Epstein-Barr Virus Nuclear Antigens; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D010957:Plasmids; D013045:Species Specificity; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics
D000956:Antigens, Viral; C529274:EBNA-6 protein, human herpesvirus 4; D019309:Epstein-Barr Virus Nuclear Antigens; D001616:beta-Galactosidase