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B/Rel Site in Intron 1 Cooperates with Proximal Promoter Elements to Mediate TNF-
-Induced Transcription of the Human Polymeric Ig Receptor1
Laboratory for Immunohistochemistry and Immunopathology, Institute of Pathology, University of Oslo, Rikshospitalet, Oslo, Norway
Secretory Abs constitute the first line of specific immune defense
at mucosal surfaces. Such Abs are generated by the active transport of
polymeric Ig (pIg) across secretory epithelia mediated by the pIgR,
also known as transmembrane secretory component (SC). The
proinflammatory cytokine TNF-
is a key mediator of host responses to
infections, and it can stimulate protein synthesis-dependent
transcriptional up-regulation of pIgR/SC in the HT-29 intestinal
adenocarcinoma cell line. By reporter gene assay we identified a
novel TNF--responsive region located within a 748-bp fragment in
intron 1 of the human pIgR/SC gene which depended on an
NF-
B/Rel site for full responsiveness. EMSAs demonstrated
preferential binding of the NF-B/Rel family member p65 (RelA) to
this DNA element after TNF- stimulation, with weaker and more
delayed binding of p50. Furthermore, the TNF--responsive region in
intron 1 required cooperation with DNA elements located in the proximal
promoter region of the gene. Mutational analysis demonstrated that an
IFN-stimulated response element near the transcriptional start site in
exon 1 was involved in the TNF- responsiveness. Thus, DNA elements
located >4 kb apart were found to cooperate in TNF--induced pIgR/SC
up-regulation. The intronic TNF--responsive enhancer overlapped with
a recently identified IL-4-responsive enhancer. Several intronic DNA
elements found to be functionally important in the human gene are
highly conserved between the human and mouse pIgR/SC genes,
suggesting the presence of a conserved cytokine-responsive enhancer
region.
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