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(Received for publication, February 29, 1996, and in revised form, April 24, 1996)
From the Imperial Cancer Research Fund Laboratories, Institute of
Molecular Medicine, University of Oxford, John Radcliffe Hospital,
Oxford OX3 9DU, United Kingdom
Expression of DNA topoisomerase II
Volume 271, Number 28,
Issue of July 12, 1996
pp. 16741-16747
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
Gene Promoter in
Confluence-arrested Cells
mRNA
and protein reflects the proliferative state of mammalian cell lines
and tissues with high levels in actively cycling cells but marked
down-regulation during serum deprivation or cell density-induced growth
arrest. Using stably integrated gene fusions comprising the human
topoisomerase II promoter with a growth hormone reporter gene, we
have localized elements required for the differential activity of the
topoisomerase II promoter in proliferating and confluence-arrested
cells. Deletion analysis localized the region of the promoter that
responded to changes in the cellular growth state to between 
101 and
144 base pairs. Mutation analysis identified an inverted CCAAT box
(ICB) located at 108 to 104 as necessary for promoter
down-regulation in confluence-arrested cells, while several other
potential cis-acting elements, including four additional
ICBs, were shown not to be required. The critical ICB was recognized
in vitro by the CCAAT box binding factor, NF-Y, with levels
of binding activity higher in extracts from proliferating cells than
from confluence-arrested cells. We conclude that the differential
regulation of topoisomerase II gene expression in cycling and
confluence-arrested cells is mediated, at least in part, through
proliferation-specific binding of factors to an ICB element in the gene
promoter.
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