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(Received for publication, January 26, 1996, and in revised form, July 1, 1996)
From Glucagon gene transcription in the endocrine
pancreas is regulated by at least four cis-acting DNA
control elements. We showed previously that G1 is critical for alpha
cell-specific expression. G1 contains three AT-rich sequences important
for promoter function, which represent candidate binding sites for
homeodomain transcription factors. Performing reverse
transcription-polymerase chain reaction amplifications with degenerate
oligonucleotide primers homologous to the Antennapedia
homeobox, cDNA clones corresponding to the caudal-related gene cdx-2/3 were predominantly
obtained from glucagon-producing cells and primary non-beta cells. From
RNase protection and polymerase chain reaction analyses,
cdx-2/3 turned out to be the only
caudal-related gene that is expressed at significant levels
in cells of the endocrine pancreas. Cdx-2/3 binds with high affinity to
an AT-rich motif of G1, which matches the consensus binding site of
caudal-related proteins. In the glucagon-producing hamster
cell line InR1G9, Cdx-2/3 is a subunit of complex B3 formed on G1.
Alternative splicing generates two cdx-2/3 transcripts in
islet cells, coding for a full-length protein and an amino-terminally
truncated isoform. Although both isoforms bind G1 with similar
affinity, only the full-length Cdx-2/3 A protein activates glucagon
gene transcription in non-glucagon-producing cells, transcriptional
activation being dose-dependent. We therefore conclude that
the caudal-related gene cdx-2/3 is implicated
in the transcriptional control of glucagon gene expression in the alpha
cells of the islets of Langerhans.
Volume 271, Number 46,
Issue of November 15, 1996
pp. 28984-28994
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
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