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(Received for publication, December 4, 1996, and in revised form, April 8, 1997)
From the 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
Volume 272, Number 26,
Issue of June 27, 1997
pp. 16688-16699
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
,
and§
Gastrointestinal Unit and
§ Hematology-Oncology Unit, Massachusetts General Hospital,
Harvard Medical School, Boston, Massachusetts 02114
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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