Nucleic Acids Research, Vol 24, Issue 12 2311-2317, Copyright © 1996 by Oxford University Press
LS Coles, P Diamond, F Occhiodoro, MA Vadas and MF Shannon
The human granulocyte-macrophage colony stimulating factor (GM-CSF) gene
promoter binds a sequence-specific single-strand DNA binding protein termed
NF-GMb. We previously demonstrated that the NF-GMb binding sites were
required for repression of tumor necrosis factor- alpha (TNF-alpha)
induction of the proximal GM-CSF promoter sequences in fibroblasts. We now
describe the isolation of two different cDNA clones that encode cold shock
domain (CSD) proteins with NF-GMb binding characteristics. One is identical
to the previously reported CSD protein dbpB and the other is a previously
unreported variant of the dbpA CSD factor. This is the first report of CSD
factors binding to a cytokine gene. Nuclear NF-GMb and expressed CSD
proteins have the same binding specificity for the GM-CSF promoter and
other CSD binding sites. We present evidence that CSD factors are
components of the nuclear NF-GMb complex. We also demonstrate that
overexpression of the CSD proteins leads to complete repression of the
proximal GM-CSF promoter containing the NF-GMb/CSD binding sites.
Surprisingly, we show that CSD overexpression can also directly repress a
region of the promoter which apparently lacks NF-GMb/CSD binding sites.
NF-GMb/CSD factors may hence be acting by two different mechanisms. We
discuss the potential importance of CSD factors in maintaining strict
regulation of the GM-CSF gene.
ARTICLES
Cold shock domain proteins repress transcription from the GM-CSF promoter
Division of Human Immunology, Hanson Centre for Cancer Research, Institute of Medical and Veterinary Science, Adelaide, South Australia, Australia.
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