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Differential transcriptional and posttranscriptional regulation of gene
expression of the colony-stimulating factors by interleukin-1 and fetal
bovine serum in murine fibroblasts
JH Falkenburg, MA Harrington, RA de Paus, WK Walsh, R Daub, JE Landegent and HE Broxmeyer
Department of Medicine (Hematology/Oncology), Indiana University School of
Medicine, Indianapolis.
Colony-stimulating factors (CSF) are important factors in the proliferation
and differentiation of hematopoietic progenitor cells (HPC), and in the
survival and activation of mature blood cells. Interleukin-1 (IL-1)
combined with fetal bovine serum (FBS) strongly induces the expression of
macrophage-CSF (M-CSF), granulocyte-CSF (G- CSF), and
granulocyte-macrophage-CSF (GM-CSF) in fibroblasts. Here, we report on the
regulation of CSF gene expression in murine fibroblasts following IL-1 and
FBS stimulation. We demonstrate that 10T1/2 murine fibroblasts induced by
FBS or IL-1 accumulate M-CSF messenger RNA (mRNA). G-CSF mRNA expression
was induced by IL-1, and not by FBS. For GM-CSF expression, induction with
both FBS and IL-1 was required. Blocking studies with actinomycin-D showed
that active transcription is essential for accumulation of all three CSF
mRNAs. After blocking protein synthesis with cycloheximide, IL-1- or
FBS-induced M-CSF expression and IL-1 plus FBS-induced GM-CSF expression
still occurred and was increased. IL-1-induced G-CSF expression was
completely prevented in these cells by pretreatment with cycloheximide,
illustrating that, for this effect, intermediate protein synthesis was
required. The half-lives of M-CSF transcripts were not substantially
altered by addition of IL-1, FBS, or FBS plus IL-1. Using nuclear run- on
assays, we demonstrated that the transcription rate of M-CSF was increased
up to 20-fold by the addition of FBS, IL-1, or FBS plus IL-1. After
blocking protein synthesis with cycloheximide, IL-1-or FBS- induced
increase in M-CSF transcription rate was also observed. GM-CSF
transcription increased up to fourfold after induction with FBS or IL- 1.
G-CSF transcription rate was not altered by FBS or IL-1. Our results
indicate that M-CSF expression induced by FBS or IL-1 in these fibroblasts
is primarily regulated at the transcriptional level. GM-CSF expression
appears to be regulated both transcriptionally and posttranscriptionally,
and G-CSF expression is regulated mainly at the posttranscriptional level.
Volume 78,
Issue 3,
pp. 658-665,
08/01/1991
Copyright © 1991 by The American Society of Hematology
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