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  • Title: A CACCC box-like cis-regulatory element of the Epstein-Barr virus ED-L2 promoter interacts with a novel transcriptional factor in tissue-specific squamous epithelia.
    Author: Nakagawa H, Inomoto T, Rustgi AK.
    Journal: J Biol Chem; 1997 Jun 27; 272(26):16688-99. PubMed ID: 9195985.
    Abstract:
    The Epstein-Barr (EBV) virus induces a lytic state after infecting epithelial cells. Subsequently, there is infection of B lymphocytes with two types of cycles, latent and lytic. Apart from linkage of the EBV latent membrane protein-1 (LMP-1) with benign and malignant conditions of squamous epithelial cells, little is known about other EBV gene products that may be important in these processes as well as cellular transcriptional factors that regulate EBV gene expression in these epithelial cells. The EBV ED-L2 promoter, an early lytic cycle promoter, is located upstream of a transcription start site for a short open reading frame designated BNLF2 and just downstream of the BNLF1 (LMP-1) open reading frame. We have previously used the EBV ED-L2 promoter to target oncogenes in transgenic mice, resulting in tissue-specific expression in the tongue, esophagus, forestomach, and skin, all sharing stratifying squamous epithelia, alternatively called keratinocytes. In the present study, we have functionally dissected the ED-L2 promoter by making deletion constructs fused to the luciferase reporter gene with transient transfections into squamous and nonsquamous epithelial cell lines as well as B lymphocytes. A CACCC box-like cis-regulatory element has been identified that is located between -218 and -187 base pairs of the ED-L2 promoter that confers significant promoter activity only in squamous epithelial cells. This cis-regulatory element is active in a heterologous minimal herpes simplex virus thymidine kinase promoter reporter gene construct when transfected into squamous epithelial cells but not in nonsquamous epithelial cells. DNA gel mobility shift assays have led to the identification of DNA-protein complexes that bind the CACCC box-like element. One of these proteins is a novel transcriptional factor that is uniquely active in stratified squamous epithelial cells, designated as keratinocyte specific factor (KSF). KSF may be related to Sp1 but appears to be distinct from Sp1. In addition, KSF may interact with related or identical cis-regulatory elements found in human papillomavirus-11 E6 and cytokeratin K3 promoters that are active in keratinocytes. In aggregate, KSF may be important in the transcriptional regulation of viral and eukaryotic genes in keratinocytes.
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