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Involvement of a nuclear factor-kappa B-like protein in induction of the
macrophage colony-stimulating factor gene by tumor necrosis factor
H Yamada, S Iwase, M Mohri and D Kufe
Laboratory of Clinical Pharmacology, Dana-Farber Cancer Institute, Harvard
Medical School, Boston, MA 02115.
The macrophage colony-stimulating factor (M-CSF) is required for the growth
and differentiation of mononuclear phagocytes. Previous studies have
demonstrated that tumor necrosis factor (TNF) induces transcription of the
M-CSF gene in human myeloid cells. The present work examined the effects of
TNF on cis-acting elements in the M-CSF promoter. Deleted forms of the
M-CSF promoter were linked to the chloramphenicol acetyltransferase (CAT)
gene and transfected by electroporation into HL-60 promyelocytic leukemia
cells. The results demonstrate that an enhancer responsive to TNF
stimulation is located between positions -406 and -344 upstream to the
transcription start site. The fragment from positions -419 to -304 was
cloned 5' to the heterologous thymidine kinase (TK) promoter and linked to
the CAT gene. Both orientations of this fragment enhanced TK-promoter
activity in TNF- treated HL-60 cells. The results of gel mobility shift
assays with the - 419 to -304 fragment demonstrate binding of a
constitutive nuclear protein. A TNF-inducible protein also bound to this
fragment and resulted in a different mobility pattern. Binding of the
TNF-induced nuclear protein to the -419 to -304 fragment was inhibited by
an oligonucleotide containing the nuclear factor-kappa B (NF-kappa B)
consensus sequence. DNA footprinting demonstrated protection of an NF-
kappa B binding site at positions -377 to -368. Methylation interference
assays showed that the TNF-induced protein made contact points with guanine
residues in the same NF-kappa B sequence. Taken together, the findings
provide evidence for involvement of an NF-kappa B-like factor in
transcriptional regulation of the M-CSF gene.
Volume 78,
Issue 8,
pp. 1988-1995,
10/15/1991
Copyright © 1991 by The American Society of Hematology
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