title: Negative regulation of the BZLF1 promoter of Epstein-Barr virus.
abstract: The Epstein-Barr virus BZLF1 gene product (ZEBRA) is a transcriptional activator whose expression in latently infected B cells is sufficient to induce the viral lytic cycle. Since there is no transcription of BZLF1 during latency, we carried out experiments to determine whether cis-acting negative elements in the BZLF1 promoter contribute to the lack of expression during this phase of the virus cycle. A series of deletion plasmids encompassing positions -551 to +14 of the BZLF1 promoter region were constructed and tested for the ability to drive chloramphenicol acetyltransferase (CAT) gene expression in the absence of inducing agents such as 12-O-tetradecanoylphorbol-13-acetate (TPA) and anti-immunoglobulin. Expression from the intact 551-bp region was very weak in most of the cell lines tested, but deletion of 165 bp from the 5' end caused a sevenfold increase in expression of CAT. Within these 165 bp, a minimal 48-bp region was sufficient to down regulate the expression of a simian virus 40/CAT fusion plasmid. The 48-bp negative element consists of 7-bp dyad symmetry elements separated by 27 bp. The rightmost half of the dyad symmetry element partially overlaps a region which has a 14-of-15-bp homology to the human cytoskeletal gamma-actin promoter.
authors.affiliation: Department of Molecular Biology, Princeton University, New Jersey 08544-1014.
authors.forename: E A
authors.initials: EA
authors.lastname: Montalvo
authors.forename: Y
authors.initials: Y
authors.lastname: Shi
authors.forename: T E
authors.initials: TE
authors.lastname: Shenk
authors.forename: A J
authors.initials: AJ
authors.lastname: Levine
mesh_terms: D000818:Animals; D001483:Base Sequence; D004268:DNA-Binding Proteins / Q000235:genetics*; D015967:Gene Expression Regulation, Viral; D006367:HeLa Cells; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry; D011401:Promoter Regions, Genetic; D011518:Proto-Oncogene Proteins / Q000235:genetics; D016760:Proto-Oncogene Proteins c-fos; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D015534:Trans-Activators / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics; D014764:Viral Proteins
chemical_list: C063676:BZLF1 protein, Herpesvirus 4, Human; D004268:DNA-Binding Proteins; D009841:Oligonucleotides; D011518:Proto-Oncogene Proteins; D016760:Proto-Oncogene Proteins c-fos; D015534:Trans-Activators; D014157:Transcription Factors; D014764:Viral Proteins