Comparison of the genomic organization and promoter function for human interleukin-8 receptors A and B

Sunil K Ahuja, Ajay Shetty, H. Lee Tiffany, Philip M. Murphy

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

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 ~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.

Original languageEnglish (US)
Pages (from-to)26381-26389
Number of pages9
JournalJournal of Biological Chemistry
Volume269
Issue number42
StatePublished - Oct 21 1994
Externally publishedYes

Fingerprint

Interleukin-8B Receptors
Interleukin-8 Receptors
Exons
Base Pairing
Neutrophils
Genes
Messenger RNA
Polyadenylation
Chloramphenicol O-Acetyltransferase
Gene encoding
Jurkat Cells
5' Untranslated Regions
Alternative Splicing
Transcription
Chromosomes
Interleukin-8
Reporter Genes
Chemokines
Open Reading Frames
Cells

ASJC Scopus subject areas

  • Biochemistry

Cite this

Comparison of the genomic organization and promoter function for human interleukin-8 receptors A and B. / Ahuja, Sunil K; Shetty, Ajay; Tiffany, H. Lee; Murphy, Philip M.

In: Journal of Biological Chemistry, Vol. 269, No. 42, 21.10.1994, p. 26381-26389.

Research output: Contribution to journalArticle

Ahuja, Sunil K ; Shetty, Ajay ; Tiffany, H. Lee ; Murphy, Philip M. / Comparison of the genomic organization and promoter function for human interleukin-8 receptors A and B. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 42. pp. 26381-26389.
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