Developmental changes of GM-CSF gene inducibility in embryonal carcinoma cells.
Murine embryonal carcinoma (EC) P19 cells, a tissue culture model of early embryonic development, failed to produce cytokines, such as interleukin-3 (IL-3), IL-4, granulocytemacrophage colony stimulating factor (GM-CSF) and interferon-beta (IFN-beta) at the mRNA level. Differentiation induced by retinoic acid (RA) released this repression to produce some cytokines. GM-CSF and IFN-beta genes were expressed in response to PMA/A23187, poly(I):poly(C), IL-1 alpha, forskolin, or LPS stimulation in differentiated P19 cells, whereas IL-3 and IL-4 genes were not expressed. To elucidate the mechanism of the GM-CSF gene induction after differentiation, we transfected a series of 5' deletion mutants of the mouse GM-CSF promoter fused to the bacterial CAT gene. The 740-bp fragment of the 5'-flanking region mediated the positive response. Deletion analysis revealed that the 5' boundary region of the DNA element required for activation lies between positions -95 and -84 and the region upstream of position -95 appears inhibitory. These results indicate that the maturation of the transcriptional machinery after differentiation results in the activation of the GM-CSF gene.
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; D015152:Blotting, Northern; D002454:Cell Differentiation / Q000235:genetics; D054278:Embryonal Carcinoma Stem Cells; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects / Q000502:physiology*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000096:biosynthesis / Q000235:genetics*; D016899:Interferon-beta / Q000235:genetics; D007377:Interleukin-3 / Q000235:genetics; D015847:Interleukin-4 / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D014411:Neoplastic Stem Cells; D011401:Promoter Regions, Genetic; D017384:Sequence Deletion; D015533:Transcriptional Activation
D007377:Interleukin-3; D015847:Interleukin-4; D016899:Interferon-beta; D016178:Granulocyte-Macrophage Colony-Stimulating Factor