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J. Biol. Chem., Vol. 269, Issue 20, 14784-14791, May, 1994
HM Guardiola-Diaz, C Boswell and AF Seasholtz
Membrane depolarization is a critical element of neuronal signaling. In
this study, the biochemical and molecular mechanisms involved in
transcriptional regulation of the corticotropin-releasing hormone (CRH)
gene by depolarization were investigated. In PC-12 cells, potassium-
induced membrane depolarization increased expression of a CRH-reporter
construct in a cAMP-dependent manner. This synergistic activation was
mediated via calcium influx, predominantly via L-type calcium channels, and
calmodulin. RNase protection assays demonstrated increased levels of
CRH-reporter transcripts in stably transfected cells after treatment with
cAMP and potassium, with the induced transcripts initiating at the major
transcription initiation site of the human CRH gene. At the genomic level,
the CRH cAMP-responsive element conferred both positive cAMP and
synergistic cAMP/depolarization regulation to a heterologous promoter.
Additionally, DNase I protection assays demonstrated similar nuclear
protein/DNA binding profiles across the cAMP-responsive element after
treatment of PC-12 cells with potassium or potassium/cAMP. These results
support a model in which the protein(s) binding to the cAMP- responsive
element integrates signals initiated by multiple pathways (cAMP and
calcium) and transmits that integrated signal to the basal transcription
machinery, resulting in increased levels of gene expression.
The cAMP-responsive element in the corticotropin-releasing hormone gene mediates transcriptional regulation by depolarization
Department of Biological Chemistry, University of Michigan, Ann Arbor 48109.
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