Journal Article

Distinct Roles of E2F Recognition Sites as Positive or Negative Elements in Regulation of the DNA Polymerase α 180 kDa Catalytic Subunit Gene Promoter during <i>Drosophila</i> Development

Masamitsu Yamaguchi, Yuko Hayashi, Fumiko Hirose, Yoshio Nishimoto and Akio Matsukage

in Nucleic Acids Research

Volume 25, issue 19, pages 3847-3854
Published in print October 1997 | ISSN: 0305-1048
Published online October 1997 | e-ISSN: 1362-4962 | DOI: https://dx.doi.org/10.1093/nar/25.19.3847
Distinct Roles of E2F Recognition Sites as Positive or Negative Elements in Regulation of the DNA Polymerase α 180 kDa Catalytic Subunit Gene Promoter during Drosophila Development

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The transcription factor E2F plays a key role in transcriptional control during the growth cycle of higher eukaryotic cells. The promoter region of the Drosophila DNA polymerase α 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 α 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 α 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 α 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 α gene promoter during Drosophila development.

Journal Article.  5602 words.  Illustrated.

Subjects: Chemistry ; Biochemistry ; Bioinformatics and Computational Biology ; Genetics and Genomics ; Molecular and Cell Biology

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