Evidence for repressional role of an inverted CCAAT box in cell cycle- dependent transcription of the human DNA topoisomerase IIα gene

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Abstract

Expression of DNA topoisomerase IIα (topo IIα) is cell cycle-regulated at both the transcriptional and the post-transcriptional levels. In order to identify cis-acting elements responsible for transcriptional regulation during the cell cycle, we investigated NIH/3T3 cells stably transfected with luciferase reporter plasmids containing various lengths of the human topo IIα gene promoter. Serum-deprived cells expressed low levels of luciferase, and following serum-induced cell cycle reentry luciferase levels were gradually elevated 2-fold. During S phase, a steep 3-fold increase in luciferase activity was seen, reaching its maximum approximately 22 h after serum addition. This pattern was observed with both a full-length (nucleotides (nt) -295 to +90] and a deletion (nt -90 to +90) promoter construct. In contrast, when testing a deletion construct (nt -51 to +90) lacking the first inverted CCAAT box (ICB1) the S phase-specific induction was absent. Mutation of ICB1 revealed that it had a repressive character, since luciferase levels rose rapidly to maximal levels immediately following serum addition. Furthermore, electrophoretic mobility shift assays demonstrated a marked decrease in ICB1 binding activity following serum addition. Together, this suggests a role of ICB1 in cell cycle-dependent repression of topo Hα transcription.

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Falck, J., Jensen, P. B., & Sehested, M. (1999). Evidence for repressional role of an inverted CCAAT box in cell cycle- dependent transcription of the human DNA topoisomerase IIα gene. Journal of Biological Chemistry, 274(26), 18753–18758. https://doi.org/10.1074/jbc.274.26.18753

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