Nucleic Acids Research, Vol 25, Issue 19 3847-3854, Copyright © 1997 by Oxford University Press
M Yamaguchi, Y Hayashi, F Hirose, Y Nishimoto and A Matsukage
The transcription factor E2F plays a key role in transcriptional control
during the growth cycle of higher eukaryotic cells. The promoter region of
the DrosophilaDNA polymerase alpha 180 kDa catalytic subunit gene contains
three E2F recognition sequences located at positions -353 to -342 (E2F site
1), -21 to -14 (E2F site 2) and -12 to -5 (E2F site 3) with respect to the
transcription initiation site. Various base substitutions were generated in
each or all of the three E2F sites in vitro to allow examination of their
effects on E2F binding and promoter function in cultured Kc cells as well
as in living flies. Glutathione S-transferase (GST)-E2F and GST-DP fusion
proteins were found to cooperate in binding to the three E2F sites in the
DNA polymerase alpha gene promoter in vitro. In contrast, an E2F-specific
activity detected in nuclear extracts of Kc cells showed little affinity
for E2F site 1 but strong binding to sites 2 and 3. Transient expression of
Drosophila E2F in Kc cells activated the DNA polymerase alpha gene promoter
and the target sites for activation coincided with E2F sites 2 and 3.
However, analyses with transgenic flies indicate that E2F site 3 functions
positively in terms of DNA polymerase alpha gene promoter activity, while
E2F sites 1 and 2 rather have a negative control function. Thus E2F sites
play distinct roles as positive or negative elements in regulation of the
DNA polymerase alpha gene promoter during Drosophila development.
ARTICLES
Distinct roles of E2F recognition sites as positive or negative elements in regulation of the DNA polymerase alpha 180 kDa catalytic subunit gene promoter during Drosophila development
Laboratory of Cell Biology, Aichi Cancer Center Research Institute, Chikusa-ku, Nagoya 464, Japan. myamaguc@aichi-cc.pref.aichi.jp
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