Cloning and characterization of the human neutrophil-activating peptide (ENA-78) gene.
The neutrophil-activating peptide (ENA-78) is an inflammatory chemokine which is produced concomitantly with interleukin-8 (IL-8) in response to stimulation with either interleukin-1 (IL-1 beta) or tumor necrosis factor-alpha (TNF-alpha). We have identified a full-length ENA-78 cDNA and isolated its genomic clone. The gene was found to consist of four exons and three introns and its structure resembles the IL-8 gene. The human ENA-78 gene was mapped to chromosome 4q13-q21, the same locus as several other inflammatory cytokine genes. The transcription initiation site was mapped to a position 96 base pairs (bp) upstream from the translation initiation site. A fusion gene containing 125 bp upstream of exon 1 linked to a luciferase reporter gene was expressed in the human embryonic 293 cell line. The expression of the reporter gene was induced by TNF-alpha, IL-1 beta, or phorbol 12-myristate 13-acetate. The 125-bp promoter region contained the cis-regulatory elements for enhancer binding protein-like factor (C/EBP) and the nuclear factor (NF-kappa B). Transfection of 293 cells with deletion mutants demonstrated that the NF-kappa B element, but not the C/EBP site, is sufficient for expression and induction by either TNF-alpha or IL-1 beta. In contrast, the IL-8 gene requires both elements. This report demonstrates that ENA-78 and IL-8 genes shared great similarity in genomic structure and chromosome location. However, these two genes may be regulated by distinct mechanisms.
Chang
M S
MS
Department of Developmental Biology, Amgen Inc., Thousand Oaks, California 91320.
McNinch
J
J
Basu
R
R
Simonet
S
S
D000595:Amino Acid Sequence; D001483:Base Sequence; D002478:Cells, Cultured; D054365:Chemokine CXCL5; D019743:Chemokines, CXC; D002874:Chromosome Mapping; D002894:Chromosomes, Human, Pair 4; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005091:Exons / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects; D017930:Genes, Reporter; D015894:Genome, Human; D006801:Humans; D017403:In Situ Hybridization; D007375:Interleukin-1 / Q000494:pharmacology; D016209:Interleukin-8 / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000235:genetics; D007438:Introns / Q000235:genetics; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D008969:Molecular Sequence Data; D005810:Multigene Family / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
C486263:CXCL5 protein, human; D054365:Chemokine CXCL5; D019743:Chemokines, CXC; D018076:DNA, Complementary; D007375:Interleukin-1; D016209:Interleukin-8; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; D008156:Luciferases; D013755:Tetradecanoylphorbol Acetate