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Requirement for nuclear factor (NF)-kappa B p65 and NF-interleukin-6
binding elements in the tumor necrosis factor response region of the
granulocyte colony-stimulating factor promoter
SM Dunn, LS Coles, RK Lang, S Gerondakis, MA Vadas and MF Shannon
Division of Human Immunology, Hanson Centre for Cancer Research, Institute
of Medical and Veterinary Science, Adelaide, South Australia.
Granulocyte colony-stimulating factor (G-CSF) is a hematopoietic growth
factor produced by mesenchymal and myeloid cells following activation by
inflammatory stimuli. It has previously been shown that a region of the
G-CSF promoter, (-200 to -165) containing the decanucleotide CK-1 element
and two repeated sequences that resemble nuclear factor (NF)- interleukin-6
(IL-6) binding sites, is required for activation of the G- CSF gene by
tumor necrosis factor-alpha (TNF-alpha) and IL-1 beta. We now show that the
NF-kappa B p65 protein can bind to and activate this TNF response region.
There are several unusual features of this p65 interaction with the TNF
response region. First, NF-kappa B p65 but not the related NF-kappa B p50
binds to the CK-1 element and a p50/65 hybrid protein that relies on the
p50 rel homology domain for DNA binding does not transactivate the TNF
response region. Second, p65 transactivation of this region is cell
specific and requires not only its own binding site but also the NF-IL6
consensus sites. NF-IL6 also binds to the TNF response region of the G-CSF
promoter. Electrophoretic mobility shift studies show that p65 and NF-IL6
can bind cooperatively to the TNF response region. The ability of this
region to respond to TNF-alpha or p65 is correlated with the ability to
form the p65/NF-IL6 ternary complex.
Volume 83,
Issue 9,
pp. 2469-2479,
05/01/1994
Copyright © 1994 by The American Society of Hematology
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