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(Received for publication, June 9, 1995)
Several different oncogenes and growth factors promote G
phase progression. Cyclin D1, the regulatory subunit of several
cyclin-dependent kinases, is required for, and capable of shortening,
the G phase of the cell cycle. The present study
demonstrates that transforming mutants of p21 (Ras Val-12, Ras Leu-61) induce the cyclin D1 promoter in
human trophoblasts (JEG-3), mink lung epithelial (Mv1.Lu), and in
Chinese hamster ovary fibroblast cell lines. Site-directed mutagenesis
of AP-1-like sequences at -954 abolished
p21
-dependent activation of cyclin D1
expression. The AP-1-like sequences were also required for activation
of the cyclin D1 promoter by c-Jun. In electrophoretic mobility shift
assays using nuclear extracts from cultured cells and primary tissues,
several AP-1 proteins (c-Jun, JunB, JunD, and c-Fos) bound the cyclin
D1 -954 region. Cyclin D1 promoter activity was stimulated by
overexpression of mitogen-activated protein kinase
(p41
) or c-Ets-2 through the proximal 22 base
pairs. Expression of plasmids encoding either dominant negative MAPK
(p41
) or dominant negatives of ETS activation
(Ets-LacZ), antagonized MAPK-dependent induction of cyclin D1 promoter
activity. Epidermal growth factor induction of cyclin D1 transcription,
through the proximal promoter region, was antagonized by either
p41
or Ets-LacZ, suggesting that ETS functions
downstream of epidermal growth factor and MAPK in the context of the
cyclin D1 promoter. The activation of cyclin D1 transcription by
p21
provides evidence for cross-talk between the
p21
and cell cycle regulatory pathways.
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