Differences in the extent of activation of Epstein-Barr virus replicative gene expression among four nonproducer cell lines stably transformed by oriP/BZLF1 plasmids.
Lymphoid cell lines were established which stably carry the Epstein-Barr viral (EBV) BZLF1 gene on an extrachromosomal plasmid. These lines, which spontaneously synthesize the BZLF1 gene product, ZEBRA, were examined for expression of EBV genes which were activated by ZEBRA. Cell lines which acquired oriP plasmids without BZLF1 served as controls. The extent of activation differed among derivatives of four cell lines. X50-7 cells, which harbor a standard latent EBV, could be induced by ZEBRA to produce transforming virus; a cellular subclone of this line was induced to express EBV late antigens but did not release transforming virus. In two other cell lines, Raji and ER, ZEBRA activated only a group of early antigens. Using immunofluorescence and immunoblotting with monoclonal antibodies and Northern analysis five EBV early genes were shown to be induced in cells stably transformed by oriP/BZLF1 plasmids. ZEBRA itself was activated; thus BZLF1 is autostimulatory. Four other activated genes were components of the diffuse (EA-D) and restricted (EA-R) early antigens (BMRF1, BMLF1, BHRF1, and BORF2). Stable cell lines with extrachromosomal BZLF1 expression vectors will ultimately be useful in a variety of experiments designed to study regulation of this gene, to analyze the effects of mutations on ZEBRA protein function, and to define the full spectrum of viral and cellular genes which are activated by and interact with the ZEBRA protein.
Gradoville
L
L
Department of Epidemiology and Public Health, Yale University School of Medicine, New Haven, Connecticut 06510.
Grogan
E
E
Taylor
N
N
Miller
G
G
D002460:Cell Line; D002472:Cell Transformation, Viral; D004261:DNA Replication; D004279:DNA, Viral / Q000096:biosynthesis*; D004352:Drug Resistance, Microbial / Q000235:genetics; D015967:Gene Expression Regulation, Viral; D005816:Genetic Complementation Test; D004854:Herpesvirus 4, Human / Q000235:genetics* / Q000254:growth & development; D006801:Humans; D009154:Mutation; D010957:Plasmids / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics*; D014158:Transcription, Genetic; D014779:Virus Replication / Q000235:genetics*
D004279:DNA, Viral; D012333:RNA, Messenger; D015534:Trans-Activators