Nucleic Acids Research, Vol 25, Issue 14 2759-2765, Copyright © 1997 by Oxford University Press
MI Darville and GG Rousseau
There is one class of genes whose expression increases at the G1/S
transition of the cell cycle. One of these genes codes for 6-
phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFK-2), an enzyme that
controls glycolysis. The cell-cycle regulation of the PFK-2 gene depends on
a binding site for the transcription factor E2F located at the 5'end of the
first exon and involves not only a transcriptional, but also a
post-transcriptional, mechanism. We have investigated this mechanism by
studying in Rat-1 fibroblasts mature and immature mRNAs from the endogenous
PFK-2 gene and from stably expressed transgenes containing PFK-2 gene
regions. An increase in precursor mRNA half-life and processing took place
at the G1/S transition. Transgenes with a mutated E2F binding site or with
mutated splice sites lost the regulation by serum, indicating that both an
intact E2F binding site and an efficient splicing reaction are necessary
for proper mitogenic stimulation. In quiescent cells, the transgene lacking
the E2F binding site was more efficiently spliced than the wild-type
construct. These results indicate that, in the wild-type gene, precursor
mRNA splicing is blocked in G0and that this block requires the E2F binding
site. The data provide evidence for a coupling between stimulation of
promoter activity and increased mRNA splicing in the mitogenic regulation
of S phase-regulated genes.
ARTICLES
E2F-dependent mitogenic stimulation of the splicing of transcripts from an S phase-regulated gene
Hormone and Metabolic Research Unit, Louvain University Medical School and International Institute of Cellular and Molecular Pathology, 75 Avenue Hippocrate, B-1200 Brussels, Belgium.
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