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J. Biol. Chem., Vol. 278, Issue 19, 16770-16776, May 9, 2003
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From the We have previously demonstrated that ectopic
expression of E2F1 is sufficient to drive quiescent cells into S phase
and that E2F1 expression can contribute to oncogenic transformation.
Key target genes in this process include master regulators of the cell
cycle, such as cyclin E, which regulates G1
progression, and cyclin A, which is required for the initiation of DNA
synthesis. In the present work, we present novel evidence that a second
G1 cyclin, cyclin D3, is also potently activated by E2F1.
First, an estrogen receptor-E2F1 fusion protein (ER-E2F1) potently
activates the endogenous cyclin D3 mRNA upon treatment with
4-hydroxytamoxifen, which induces nuclear accumulation of the otherwise
cytosolic fusion protein. Furthermore, trans-activation of cyclin D3 by ER-E2F1 occurs even in the presence of the protein synthesis inhibitor cycloheximide and thus appears direct. Second, all of the
growth-stimulatory members of the E2F family (E2F1, -2, and -3A)
potently activate a cyclin D3 promoter reporter, whereas
growth-restraining members of the family (E2F4, -5, and -6) have little
effect. Third, recombinant E2F1 binds with high affinity to the cyclin
D3 promoter in vitro. Fourth, chromatin immunoprecipitation
assays demonstrate that endogenous E2F1 is associated with the cyclin
D3 promoter in vivo. Finally, mapping experiments localize
the essential E2F regulatory element of the cyclin D3 promoter to a
noncanonical E2F site in the promoter between nucleotides
Regulation of the Cyclin D3 Promoter by E2F1*
§,§,§,§, and§¶
Program in Molecular Oncology, H. Lee
Moffitt Comprehensive Cancer Center and Research Institute and the
§ Department of Interdisciplinary Oncology, University of
South Florida, College of Medicine, Tampa, Florida 33612
143 and
135 relative to the initiating methionine codon. We conclude that in
addition to cyclins E and A, E2F family members can also activate one
member of the D-type cyclins, further contributing to the ability of
the stimulatory E2F family members to drive cellular proliferation.
*
This work was supported by NCI, National Institutes of
Health, Grant CA78214 and American Cancer Society Grant
RSG-02-239-01-GMC (to W. D. C.) and by the H. Lee Moffitt Cancer
Center and Research Institute.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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