Characterization of sublines of Epstein-Barr virus producing HR-1 cells and its implication in virus propagation in culture.
To understand the mechanism regulating the EBV replication cycle, several sublines were obtained from HR-1 cells by the limiting dilution method. Based on their biochemical and molecular characteristics, these sublines can be categorized into two classes: the high EBV-DNA containing (H) subline and low EBV-DNA containing (L) subline. The amount of EBV proteins, such as EBV polymerases, EBV DNase, EAD, ZEBRA, MA, and VCA, was much higher in H sublines than in L sublines. Only 20% of cells in the H subline express those proteins. In addition to regular EBV DNA restriction enzyme fragments, additional DNA restriction enzyme fragments, as detected by different EBV DNA fragment probes, were found to be present in H sublines but not in L sublines. No BamH1 W-Z DNA fragment rearrangement, which was the primary reason for ZEBRA expression in a high EBV-DNA containing subline, Clone 5, was found in H sublines. When L sublines were treated with 12-0-tetradecanoylphorbol-13-acetate and sodium butyrate, EBV-specific proteins, including ZEBRA protein, could be induced in cells, but no virus could be detected in the medium. Thus, the lack of EBV production by L sublines is more than the simple lack of expression of ZEBRA protein. L sublines are susceptible to EBV infection and are capable of producing EBV after infection. The importance of the presence of L cells in the H subline for the propagation of EBV in culture is suggested.
Yao
G Q
GQ
Department of Pharmacology, Yale University School of Medicine, New Haven, CT, USA.
Tsai
C H
CH
Cheng
Y C
YC
D002051:Burkitt Lymphoma / Q000473:pathology / Q000821:virology; D002087:Butyrates / Q000494:pharmacology; D020148:Butyric Acid; D002460:Cell Line; D004261:DNA Replication; D004279:DNA, Viral / Q000096:biosynthesis / Q000235:genetics; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics; D015967:Gene Expression Regulation, Viral / Q000187:drug effects; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D006801:Humans; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D015534:Trans-Activators / Q000096:biosynthesis / Q000235:genetics; D014407:Tumor Cells, Cultured; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics; D014776:Virus Cultivation; D017735:Virus Latency; D014779:Virus Replication / Q000187:drug effects
C063676:BZLF1 protein, Herpesvirus 4, Human; D002087:Butyrates; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D015534:Trans-Activators; D014764:Viral Proteins; D020148:Butyric Acid; D013755:Tetradecanoylphorbol Acetate