Analysis of the cytokine-stimulated human inducible nitric oxide synthase (iNOS) gene: characterization of differences between human and mouse iNOS promoters.
Expression of human inducible nitric oxide synthase (hiNOS) is under cytokine control and is transcriptionally regulated. The hiNOS and mouse iNOS (miNOS) genes are regulated differently by cytokines. To understand better the transcriptional regulation of the hiNOS gene, the 8.3-kb hiNOS promoter was characterized. Promoter activity was evaluated by transient transfection of hiNOS luciferase constructs in A549 human alveolar type II epithelium-like cells in the presence and absence of cytokines (IFN-gamma, IL-1 beta, and TNF-alpha). Important cytokine-responsive elements are located at -3665 to -5574 bp (containing two perfectly matched AP-1 sites which are not present in miNOS promoter) and -8093 to -8296 bp (one perfectly matched NF-kappa B site) of the hiNOS promoter region. Likely, these two AP-1 sites and the upstream NF-kappa B site are important in the transcriptional induction of hiNOS by cytokines. Our data demonstrate the molecular basis for the different cytokine-stimulated characteristics of hiNOS and miNOS genes.
Pulmonary-Critical Care Medicine Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
S C
SC
Chu
J
J
Marks-Konczalik
H P
HP
Wu
T C
TC
Banks
J
J
Moss
D000818:Animals; D001483:Base Sequence; D016207:Cytokines / Q000494:pharmacology*; D004847:Epithelial Cells; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D007375:Interleukin-1 / Q000494:pharmacology; D008156:Luciferases / Q000096:biosynthesis; D051379:Mice; D008969:Molecular Sequence Data; D019001:Nitric Oxide Synthase / Q000096:biosynthesis* / Q000235:genetics*; D052247:Nitric Oxide Synthase Type II; D011401:Promoter Regions, Genetic; D011650:Pulmonary Alveoli; D012333:RNA, Messenger / Q000096:biosynthesis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D016207:Cytokines; D007375:Interleukin-1; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; D008156:Luciferases; C496319:NOS2 protein, human; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse