Epstein-Barr virus (EBV) replication and expressions of EA-D (BMRF1 gene product), virus-specific deoxyribonuclease, and DNA polymerase in EBV-activated Akata cells.
The replication of Epstein-Barr virus (EBV) and the expression of EBV early proteins were studied in the Burkitt's lymphoma cell line Akata stimulated with anti-human immunoglobulin G antibody (anti-IgG). Akata cells contained approximately 20 copies of EBV genome per cell as covalently closed, circular DNA. EBV DNA replication was observed at 6 hr and reached a maximal level at 24 hr after treatment with anti-IgG. Virion DNA was found in the culture medium at 12 hr. The kinetics of expression of BMRF1 gene product (early antigen diffuse component; EA-D) paralleled that of EBV deoxyribonuclease (DNase) and of DNA polymerase. Immunoblotting analysis showed that three polypeptides with molecular masses of 54, 52, and 49 kilodaltons (kDa) were recognized as EA-D components. The EBV DNase polypeptide was detected by immunoblotting at 53 kDa. The anti-EBV DNA polymerase antibody recognized 120- and 54-kDa polypeptides in the Akata cells. Immunoprecipitation followed by immunoblotting showed that EA-D and EBV DNase polypeptides were coimmunoprecipitated with anti-EBV DNase antibody and with anti-EA-D monoclonal antibody. These findings indicate that EA-D forms a complex with EBV DNase polypeptide. The molecules of EA-D, EBV DNase, and DNA polymerase appear to be closely associated together on the EBV replication.
Department of Pharmacology, University of Massachusetts Medical School, Worcester 01655.
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D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000096:biosynthesis / Q000276:immunology; D002051:Burkitt Lymphoma / Q000382:microbiology*; D002472:Cell Transformation, Viral; D003429:Cross Reactions; D004261:DNA Replication; D004268:DNA-Binding Proteins; D004259:DNA-Directed DNA Polymerase / Q000096:biosynthesis / Q000276:immunology; D003851:Deoxyribonucleases / Q000096:biosynthesis / Q000276:immunology; D016679:Genome, Viral; D004854:Herpesvirus 4, Human / Q000201:enzymology / Q000254:growth & development*; D007074:Immunoglobulin G / Q000276:immunology; D014407:Tumor Cells, Cultured; D014764:Viral Proteins / Q000096:biosynthesis* / Q000276:immunology
D000914:Antibodies, Viral; D000956:Antigens, Viral; D004268:DNA-Binding Proteins; C033305:Epstein-Barr virus early antigen; C083204:Epstein-Barr virus early antigen diffuse component; D007074:Immunoglobulin G; D014764:Viral Proteins; C083202:BALF5 protein, Epstein-barr virus; D004259:DNA-Directed DNA Polymerase; D003851:Deoxyribonucleases; C083203:deoxyribonuclease, Epstein-Barr virus