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J Biol Chem, Vol. 273, Issue 29, 18499-18508, July 17, 1998
From the Department of Molecular Pharmacology, University of
Göttingen, D-37070 Göttingen, Germany
Cyclic AMP stimulates insulin gene transcription
through a cAMP response element (CRE). In the present study the insulin
CRE-binding proteins and their functions were investigated. A
mutational analysis of nuclear protein binding in electrophoretic
mobility shift assays in combination with specific antisera showed that
in the CRE of the rat insulin I gene the imperfect CRE octamer-like
sequence TGACGTCC interacts weakly with CREB and overlaps with two
sequence motifs (TTGTTGAC and CCAAT) that bind winged helix-like
proteins and the transcription factor NF-Y, respectively. Transient
transfection of wild-type and mutant insulin CRE-reporter fusion genes
and the inactivation of cellular CREB or NF-Y by overexpression of the
dominant negative mutants KCREB or NF-YA29, respectively, indicate that
cAMP inducibility of the insulin CRE is mediated by CREB or closely
related proteins; however, NF-Y binding to the insulin CRE confers
constitutive, basal activity and decreases the ability of CREB to
mediate cAMP-stimulated transcription and calcium responsiveness.
Results from these studies demonstrate that NF-Y binds to the insulin
CRE and modulates the function of CREB. Together with the
nonpalindromic sequence of the CRE octamer motif, the interaction of
NF-Y with CREB may be responsible for the gene-specific transcriptional
activity of the insulin CRE and explain why it has considerable basal
activity but is less responsive to cAMP stimulation than others.
Gene-specific Transcriptional Activity of the Insulin
cAMP-responsive Element Is Conferred by NF-Y in Combination with cAMP
Response Element-binding Protein
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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