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J Biol Chem, Vol. 274, Issue 40, 28787-28793, October 1, 1999
From the Second Department of Pathology, Kobe University School of
Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe 650-0017, Japan
Cyclin D1, a G1/S cell
cycle-regulating oncogene, is known to be transcriptionally regulated
by numerous growth factors. We cloned and characterized the rat cyclin
D1 gene 5'-flanking region and, by species- and subspecies-matched
transient transfection studies, found that a basic promoter structure
with a cAMP response element and two continuous Sp1-binding sites was
crucial for the steady-state expression of the cyclin D1 gene.
Furthermore, the methylation status especially around two continuous
Sp1-binding sites was found to be an important epigenetical mechanism
determining the steady-state expression level in rat leukemic cell
lines K4D, K4DT, and K4D16. Whether or not epigenetic control of the
cyclin D1 gene existed among normal rat tissues was further examined by
high sensitivity mapping of the methylated cytosine. In normal rat
tissues, the methylated cytosines at non-CpG loci within two continuous
Sp1-binding sites were observed in uterine stromal cells of the basal
layer and found to be demethylated in the functioning layer, possibly
by a passive demethylation mechanism through cell division. Since
in the passive demethylation process Sp1-binding sites remain
methylated in a part of the cell population, methylated cytosines at
Sp1-binding sites may be essential for keeping a number of the stromal
cells in the basal layer live against estrogen-induced proliferation
that leads to either apoptosis or compaction.
Transcriptional Regulation of Rat Cyclin D1 Gene by CpG
Methylation Status in Promoter Region
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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