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Rb represses E2F-mediated transcription in part by blocking the
trans-activation domain of E2F. In addition, Rb can convert an
E2F binding site from a positive to a negative element. To examine the
effect of a Rb-DNA-bound complex on transcription, full-length Rb was
fused to the DNA binding domain of GAL4. Here, we report that GAL4-Rb
can repress transcription mediated by either Sp1, AP-1, or p53,
dependent upon the presence of both the GAL4 DNA binding domain and
GAL4 binding sites. Moreover, GAL4-Rb inhibited the activity of the
herpes simplex virus tk promoter from GAL4 binding sites
located at a distance from the promoter. In contrast, GAL4-Rb was
unable to repress basal transcription. Cotransfection of specific
cyclins and cyclin-dependent kinases or SV40 T-antigen abolished the
repressive activity of GAL4-Rb. The domains of Rb involved in mediating
the repression of transcription were mapped to regions that are
overlapping, but not identical, to those required for the interaction
with E2F. We propose that Rb can function as a general repressor of
transcription when bound to the promoter region.
Volume 270,
Number 15,
Issue of April 14, pp. 8837-8843, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
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