Three essential promoter elements mediate tumour necrosis factor and interleukin-1 activation of the granulocyte-colony stimulating factor gene

• Published in: Growth factors (Chur, Switzerland) Vol. 7; no. 3; p. 181
• Main Authors: Shannon, M F, Coles, L S, Fielke, R K, Goodall, G J, Lagnado, C A, Vadas, M A
• Format: Journal Article
• Language: English
• Published: England 1992
• Subjects: Base Sequence; Binding Sites; Burkitt Lymphoma; Cell Line; Cell Nucleus - metabolism; Chloramphenicol O-Acetyltransferase - genetics; Chloramphenicol O-Acetyltransferase - metabolism; Fibroblasts - drug effects; Fibroblasts - physiology; Gene Expression Regulation - drug effects; Granulocyte Colony-Stimulating Factor - genetics; Humans; Interleukin-1 - pharmacology; Kinetics; Lung; Molecular Sequence Data; Mutagenesis, Site-Directed; NF-kappa B - metabolism; Nuclear Proteins - metabolism; Oligodeoxyribonucleotides; Plasmids; Promoter Regions, Genetic - drug effects; RNA - genetics; RNA - isolation & purification; RNA, Messenger - genetics; Transfection; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha - pharmacology
• ISSN: 0897-7194
• DOI: 10.3109/08977199209046923

Granulocyte-colony stimulating factor (G-CSF) is a haemopoietic growth factor produced by mesenchymal cells but not T lymphocytes after stimulation with specific cytokines or mitogens. A 330 bp promoter fragment of the human G-CSF gene induced reporter gene expression in human embryonic lung fibroblasts in response to tumor necrosis factor-alpha (TNF-alpha) or interleukin-1 beta (IL-1 beta). The same promoter fragment was not active in Jurkat T cells nor did it respond to phorbol ester in either cell type. At least three distinct elements, the CK-1 sequence, a decanucleotide present in haemopoietic growth factor genes, an NF-IL-6 consensus sequence and a consensus octamer sequence, were essential in the G-CSF promoter for TNF-alpha and IL-1 beta response. Mutation of any of these sequences abolished promoter function. In contrast, mutation of two other consensus protein binding sequences, i.e. a Pu-1 site and a CK-2-like sequence, did not eliminate promoter function. Both the CK-1 and octamer sequences acted independently as TNF-alpha and IL-1 beta responsive elements upstream of a heterologous promoter. The response of the octamer sequence and the 330 bp promoter but not the CK-1 sequence was greater with IL-1 beta than TNF-alpha reflecting a similar response of the endogenous gene.