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Laboratory of Host Defenses, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892
CC chemokine receptor 5 (CCR5) functions physiologically as a
receptor for the leukocyte chemoattractants macrophage inflammatory
protein-1
, macrophage inflammatory protein-1ß, and RANTES, and
functions pathologically as a key cell entry coreceptor for HIV-1. The
factors that regulate CCR5 expression may be useful therapeutic targets
for HIV-1 infection. To identify nuclear regulatory factors, we have
located and functionally characterized the CCR5 gene promoter. The gene
consists of two exons separated by a 1.9-kb intron. Exon 1 contains 43
bp of the 5'-untranslated region; exon 2 contains 11 bp of the
5'-untranslated region and the complete open reading frame. Primer
extension analysis identified two adjacent transcriptional start points
(tsp) that map to the first 2 bp found in the longest known CCR5 cDNA
sequence. A TATA box is present 31 bp upstream from the first tsp. CCR5
mRNA was detected constitutively in both primary human myeloid and
lymphoid cells by Northern blot hybridization. Consistent with this,
transcription of a chloramphenicol acetyltransferase reporter gene was
constitutively activated in both transiently transfected myeloid and
lymphoid cell lines by the 80-bp gene fragment located immediately
upstream of the tsp. Deletion analysis located a strong silencer
element between nucleotides -244 and -80, and a strong enhancer
element between -486 and -244. These results suggest that the gene
region between -486 and -1 may regulate the expression of CCR5 in
monocyte/macrophages and T lymphocytes.
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