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B Regulates the Expression of the Human Complement Receptor 2 Gene1
* Department of Cellular Injury, Walter Reed Army Institute of Research, Silver Spring, MD 20910; and
Department of Medicine, Uniformed Services University of the Health Sciences, Bethesda, MD 20814
CR2 is a key regulator of the B cell response to Ag. Here we show
that NF-
B enhances the expression of the human CR2 gene. Promoter
truncation, deletion, and mutagenesis studies indicated a functional
role for a consensus NF-B promoter element, as well as a
heterogeneous nuclear ribonucleoprotein D element and an overlapping X
box/E box. By supershift analysis, the first two elements bound NF-B
p50 and p65 and heterogeneous nuclear ribonucleoprotein RNP D,
respectively. The X box/E box bound regulatory factor X5 and,
surprisingly, NF-B p50 and p65. Overexpression of NF-B p50
enhanced the activity of the CR2 promoter in B cell lines and primary B
cells, suggesting a direct role for NF-B in regulating promoter
activity. Importantly, mutation of the NF-B element or the X box/E
box rendered the promoter unresponsive to NF-B p50. Using chromatin
immunoprecipitation in live B cell lines and primary B cells, we found
that NF-B proteins p50, p65, and c-Rel bound to the genomic promoter
at two locations that overlap with the consensus NF-B element or the
X box/E box. Finally, stimuli that activate NF-B enhanced the
activity of the CR2 promoter, and LPS rapidly increased the number of
CR2 proteins on the surface of primary B cells. We propose that the
NF-B signaling pathway enhances the expression of the CR2 gene, as a
result of NF-B proteins binding to two CR2 promoter elements. Thus,
at the onset of an infection, LPS could sensitize the B cell to Ag by
enhancing the level of CR2-costimulatory molecules on the cell
surface.
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