Identification and characterization of ZIIBC, a complex formed by cellular factors and the ZII site of the Epstein-Barr virus BZLF1 promoter

Abstract

The transition from latency to lytic Epstein-Barr virus replication is dependent on the Epstein-Barr virus BZLF1 gene product. Genetic and biochemical attempts to link cellular second-messenger signaling pathways that trigger this transition with the subsequent viral gene cascade have identified functional elements within the BZLF1 promoter (Zp) that appear to bind undefined cellular transcription factors. One of these previously identified sites, ZII, has homology to consensus AP-1 and CREB binding sites, implying a role for these factors in the inductive process. We have identified and characterized ZIIBC, a ZII site binding complex that is distinct from the factors previously proposed to bind this site. Active ZIIBC was found to be present in both uninduced and chemically induced cell extracts at approximately equivalent concentrations. Analysis of the DNA sequence requirements for the binding of ZIIBC to the ZII site shows that sequences homologous to AP-1 and CREB consensus sites are necessary but not sufficient for complex formation. Although the components of ZIIBC that directly contact DNA were found to be of the same molecular masses (26 and 36 kDa) in both uninduced and chemically induced cell extracts, a slight mobility difference between DNA-protein complexes formed by these two types of extracts is observable and indicates that ZIIBC is directly affected by chemical induction. The effects of ZIIBC binding to the ZII site on expression from Zp were evaluated, and they suggest that ZIIBC plays a critical role in the regulation of Zp expression.