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Institut für Medizinische Mikrobiologie und Hygiene, Universität Regensburg, Franz-Josef-Strauß-Allee 11, D-93053 Regensburg, Germany
Epstein—Barr virus (EBV) replicates in a latent or a lytic way in the infected organism, depending on the type and level of differentiation of the host cell. The switch between latency and lytic replication was previously shown, for Burkitt's lymphoma cell lines, to depend on the viral BZLF1 gene product. Protein-DNA assays were used to identify the cis-acting elements that represent the link between regulating signal transduction pathways and the viral cascade of gene expression. Specific binding of proteins to several sites of the BZLF1 promoter during latency was shown. Induction of the lytic cycle by stimulation with 12-O-tetradecanoylphorbol 13-acetate abolished the binding of these proteins to the distal promoter (positions -227 to -551), suggesting a functional role for the down-regulation of promoter activity during latency. Computer analysis identified a multiply repeated sequence motif, HI, in this region and exonuclease III footprints confirmed that these sites act as specific protein recognition sites. Using a set of reporter plasmids we were able to demonstrate a negative regulatory effect of the HI motif in some B lymphoid cell lines, in contrast to epithelial HeLa cells. The HI silencer elements are different from other silencer elements described so far in respect of their sequence and protein-binding pattern during the activation of BZLF1.
Present address: Ludwig-Maximilians Universität München, Institut für Immunologie, Goethestraße 31, D-80336 München, Germany.
Present address: Institut für Medizinische Mikrobiologie und Hygiene, Technische Universität München, Biedersteinerstraße 29, D-80802 München, Germany.
Received 3 November 1993;
accepted 8 February 1994.
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