Nucleic Acids Research, Vol 25, Issue 24 4933-4939, Copyright © 1997 by Oxford University Press
K Korner, LA Wolfraim, FC Lucibello and R Muller
The TATA- and Inr-less promoter of the human cdc25C gene is regulated
during the cell cycle through binding of a repressor to two contiguous
promoter-proximal elements, the CDE and CHR. In this study we have
characterized in detail the region of the cdc25C promoter immediately
downstream of these elements. Several lines of evidence suggest that this
region of approximately 60 bp acts as the core promoter. This sequence: (i)
harbors most of the transcription initiation sites; (ii) possesses basal
promoter activity in vivo ; (iii) shows no stable protein binding in vivo
as indicated by genomic dimethyl sulfate and phenanthroline copper
footprinting; (iv) contains single-stranded regions in vivo as shown by
potassium permanganate footprinting; (v) is hypersensitive to DNase I
cleavage in permeabilized cells. Mutational analysis of the core promoter
revealed the presence of three sites which play a role in transcription.
Two of these sites were found to represent low affinity binding sites for
transcription factors of the Sp1 family. Mutation of these sites led to
decreased levels of transcription, while their alteration to canonical Sp1
sites impaired cell cycle regulation. Thus the transient interaction of Sp1
with the core promoter appears to be necessary for maximal transcription
without perturbing cell cycle regulation.
ARTICLES
Characterization of the TATA-less core promoter of the cell cycle- regulated cdc25C gene
Institut fur Molekularbiologie und Tumorforschung (IMT), Philipps- Universitat Marburg, Emil-Mannkopff-Strasse 2, D-35033 Marburg, Germany.
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