Regulation of human interleukin 8 gene expression and binding of several other members of the intercrine family to receptors for interleukin-8.
IL-8 is produced by a wide variety of cells in response to polyclonal mitogens and cytokines. Northern blotting analysis revealed that IL-1, TNF and PMA could induce rapid expression of IL-8 mRNA in the absence of new protein synthesis. Nuclear run-off assays using different cell types demonstrated that IL-8 mRNA expression could at least be partly due to the activation of transcription. Cloning and determination of the entire sequence of IL-8 genomic DNA enabled us to explore the functional significance of the 5'-flanking enhancer region of the IL-8 gene by employing CAT assays. The results indicated that the region spanning from -94 to -71 bp is minimally sufficient for conferring responsiveness to IL-1, TNF and PMA. Further analysis using point-mutations revealed that this region consisted of two distinct cis-elements; one being the potential binding site for NFkB-like and the other for a C/EBP-like factor. These results suggested that all three stimuli, IL-1/TNF/PMA, modulate the identical combination of nuclear factors possibly by phosphorylation. We previously reported that these three stimuli activated the same serine protein kinase which phosphorylates identical 65 kDa and 74 kDa cytosol proteins in human PBMC. This IL-1/TNF/PMA-activated protein kinase is distinct from protein kinase A, protein kinase C or casein kinase in substrate specificity; in Ca and phospholipid dependency; in cyclic nucleotide dependency; and sensitivity to protein kinase inhibitors. Taken collectively, IL-1/TNF/PMA may activate a common serine protein kinase and this protein kinase may in turn directly or indirectly modulate several nuclear factors.(ABSTRACT TRUNCATED AT 250 WORDS)
Mukaida
N
N
Cancer Treatment Division, National Cancer Institute, Frederick, Maryland 21702-1201.
Hishinuma
A
A
Zachariae
C O
CO
Oppenheim
J J
JJ
Matsushima
K
K
D001483:Base Sequence; D001667:Binding, Competitive; D016207:Cytokines / Q000378:metabolism / Q000494:pharmacology; D005786:Gene Expression Regulation; D006801:Humans; D016209:Interleukin-8 / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D011971:Receptors, Immunologic / Q000378:metabolism*; D023062:Receptors, Interleukin-8A; D012045:Regulatory Sequences, Nucleic Acid
D016207:Cytokines; D016209:Interleukin-8; D011971:Receptors, Immunologic; D023062:Receptors, Interleukin-8A