Multiple elements in the promoter of granulocyte colony-stimulating factor gene regulate its constitutive expression in human carcinoma cells.
Some human carcinoma cells constitutively produce granulocyte colony-stimulating factor (G-CSF) which stimulates the proliferation and differentiation of the progenitor cells of neutrophilic granulocytes. By introducing mouse G-CSF chromosomal gene or its promoter DNA into human carcinoma cell lines of CHU-2, SK-HEP-1, and U-87MG, it was shown that the constitutive expression of G-CSF in these carcinoma cells was due to the intrinsic activation of nuclear factors which work on the promoter region of the G-CSF gene. A series of 5' deletion mutants, linker scanning mutants, and internal deletion mutants was constructed in the promoter of mouse G-CSF gene and was introduced into human CHU-2 cells to analyze their promoter activities. These studies demonstrated that at least three regulatory elements in the promoter of the G-CSF gene are essential for the constitutive expression of G-CSF in CHU-2 cells. These elements include the consensus decanucleotide "GAGRTTCCA/CC" present on G-CSF, granulocyte/macrophage colony-stimulating factor, and interleukin 3 genes and the "ATTTGCAT" octamer transcription factor binding site. Some point mutations in these consensus sequences significantly diminished the promoter activity in CHU-2 cells.
Osaka Bioscience Institute, Japan.
M
M
Nishizawa
M
M
Tsuchiya
R
R
Watanabe-Fukunaga
S
S
Nagata
D000230:Adenocarcinoma / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D002294:Carcinoma, Squamous Cell / Q000378:metabolism; D003115:Colony-Stimulating Factors / Q000235:genetics*; D015245:Deoxyribonuclease BamHI; D015252:Deoxyribonucleases, Type II Site-Specific; D005786:Gene Expression Regulation; D005910:Glioma / Q000378:metabolism; D016179:Granulocyte Colony-Stimulating Factor; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D009369:Neoplasms / Q000378:metabolism*; D010957:Plasmids; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping; D014162:Transfection; D014407:Tumor Cells, Cultured
D003115:Colony-Stimulating Factors; D016179:Granulocyte Colony-Stimulating Factor; D015245:Deoxyribonuclease BamHI; C025651:CCCGGG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific