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J. Biol. Chem., Vol. 269, Issue 45, 27855-27862, 11, 1994
T Okazaki, U Chung, T Nishishita, S Ebisu, S Usuda, S Mishiro, S Xanthoudakis, T Igarashi and E Ogata
A rise in extracellular calcium (Ca2+e) suppresses not only secretion of
parathyroid hormone (PTH) but also expression of the PTH gene to ensure
constant plasma Ca2+ level. A nuclear protein(s) in a wide variety of cells
bound to the specific DNA elements (negative Ca2+ responsive elements,
nCaREs) in the human PTH gene in sequence-specific and Ca2+e
concentration-dependent manners. Our Southwestern cloning revealed that a
redox factor protein (ref1), which was known to activate several
transcription factors via alterations of their redox state, belonged to an
nCaRE binding protein. The level of ref1 mRNA as well as of its protein was
elevated by an increase in Ca2+e concentration. In gel shift assay,
anti-ref1 antibody eliminated formation of the nCaRE-protein complex. We
also found that there was another protein(s) interacting with nCaREs and
ref1. Further, experiments with an antisense-ref cDNA expression vector
introduced into cultured cells suggested that DNA (nCaRE)-ref1 interaction
led to Ca2+e-mediated transcriptional suppression. Thus, it is concluded
that ref1 possesses transcription repressor activity in addition to its
function as a transcriptional auxiliary protein.
A redox factor protein, ref1, is involved in negative gene regulation by extracellular calcium
Fourth Department of Internal Medicine, University of Tokyo School of Medicine, Japan.
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