Molecular basis for developmental changes of GM-CSF gene inducibility in embryonal carcinoma cells.
In a previous report, we reported that the induction of GM-CSF gene in differentiated P19 cells results from the maturation of the transcriptional machinery. Here, we identified a cis-DNA element which confers the activation of GM-CSF gene in response to PMA/A23187 stimulation in differentiated state. Analysis of the 5'-flanking region between -113 and -60 revealed two elements responsible for promotion and one for inhibition, and the overall effects led to the activation of GM-CSF gene mainly through the sequence between -95 and -73. Using the oligonucleotide between -94 and -73 as a probe in gel retardation assays, we identified a DNA-binding protein, NF-GM-P19, the binding activity of which was induced after differentiation in response to PMA/A23187 stimulation. These results indicate that the induction of GM-CSF gene after differentiation results from the maturation of the transcriptional machinery which recognizes the sequence between -95 and -73.
Heike
T
T
Department of Pediatrics, Faculty of Medicine, Kyoto University, Japan.
Nishikomori
R
R
Kawai
M
M
Tsuboi
A
A
Arai
N
N
Mikawa
H
H
D000818:Animals; D001483:Base Sequence; D000001:Calcimycin / Q000494:pharmacology; D018236:Carcinoma, Embryonal / Q000235:genetics*; D002454:Cell Differentiation; D004252:DNA Mutational Analysis; D004273:DNA, Neoplasm / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D011485:Protein Binding; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D015398:Signal Transduction; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm; D004268:DNA-Binding Proteins; D000001:Calcimycin; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D013755:Tetradecanoylphorbol Acetate