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Gene transcription can be induced by cAMP and Ca
Volume 270,
Number 15,
Issue of April 14, pp. 8860-8866, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
-dependent Phosphatase Calcineurin in Gene
Transcription That Is Stimulated by cAMP through cAMP Response Elements
through distinct protein kinases phosphorylating the
transcription factor CREB, which binds to cAMP response elements (CREs)
in various genes. Induction of gene transcription by Ca
has been shown recently to depend on the
Ca
/calmodulin-dependent protein phosphatase
calcineurin in pancreatic islet cells. This study investigates the role
of calcineurin in CRE-directed gene transcription after stimulation by
cAMP. Reporter fusion genes under the transcriptional control of CREs
were transiently transfected into the cell line HIT. Pharmacological
evidence suggests that cAMP stimulates CRE-mediated transcription
through a Ca
-dependent mechanism. The
immunosuppressive drugs cyclosporin A and FK506 inhibited CRE-mediated
transcription stimulated by cAMP. At the same concentrations they also
inhibited calcineurin phosphatase activity. Reversal of calcineurin
inhibition by rapamycin or overexpression of calcineurin led to
disinhibition of CRE-mediated gene transcription. Immunoblots with a
phosphoCREB-specific antibody showed that cyclosporin A and FK506 do
not interfere with CREB phosphorylation at serine 119 stimulated with
cAMP or membrane depolarization. These results indicate that in HIT
cells stimulation of CRE-mediated transcription depends not only on the
activity of protein kinases phosphorylating CREB but also on the
Ca
/calmodulin-dependent protein phosphatase
calcineurin that is necessary for the transcriptional competence of
phosphorylated CREB.
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