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J. Biol. Chem., Vol. 269, Issue 42, 26381-26389, Oct, 1994
SK Ahuja, A Shetty, HL Tiffany and PM Murphy
Human neutrophils are highly responsive to the chemokine interleukin-8
(IL-8) owing to high levels of expression of two related receptors encoded
by the single copy genes il8ra and il8rb located on chromosome 2q34-q35. To
identify nuclear factors that regulate the expression of IL-8 receptors, we
have first defined the organization of both genes and characterized their
functional promoters. il8ra and il8rb span approximately 4 and 12 kilobase
pairs of genomic DNA, respectively. In both cases, the open reading frame
resides on a single exon. In contrast, the 5'-untranslated regions are more
complex. For il8ra, it is formed from two exons, whereas for il8rb, seven
distinct neutrophil mRNAs are formed by alternative splicing of 11 exons.
One of the splice variants, designated IL8RB3, is the predominant form for
il8rb. Two equally abundant mRNAs for il8ra, 2.0 and 2.4 kilobases in
length, are expressed in neutrophils and arise from usage of two
alternative polyadenylation signals. Primer extension analysis identified
two major transcription start points for il8ra and 11 for il8rb. Regions
extending 300 base pairs (bp) upstream from exon 1 of il8ra and 81 bp
upstream from exon 3 of il8rb have limited sequence similarity but had
strong constitutive promoter activity when cloned upstream from a
chloramphenicol acetyltransferase-encoding reporter gene and transiently
transfected into surrogate myeloid (HL-60, and U-937) and lymphoid (Jurkat)
cell lines. Neither of these regions has sequences corresponding to classic
promoter elements. In contrast, a region 643 base pairs upstream from exon
1 of il8rb had relatively low levels of constitutive promoter activity in
all three cell environments, and a conserved TATA element is located 47 bp
upstream of the 5'-end of exon 1. Thus, despite marked differences in the
complexity of their genomic organization, il8ra and il8rb encode products
that are similar in structure, function, and the major cell type of
expression.
Comparison of the genomic organization and promoter function for human interleukin-8 receptors A and B
Laboratory of Host Defenses, NIAID, National Institutes of Health, Bethesda, Maryland 20892.
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