Journal
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Volume 380, Issue 1, Pages 22-27Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.bbrc.2008.12.186
Keywords
Topoisomerase I; Circadian rhythms; Promoter analysis; Antitumor drugs; Clock genes
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Funding
- Uehara Memorial Foundation
- Japan Private School Promotion Foundation
- Saitama Medical University Internal Grant
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To identify whether Topoisomerase I (TopoI) has autonomous circadian rhythms regulated by clock genes, we tested mouse TopoI (mTopoI) promoter oscillation in NIH3T3 cells using a real-time monitoring assay and Topol mRNA oscillations using real-time RT-PCR. Analysis of the mTopol promoter region with MatInspector software revealed two putative E-box (E1 and E2) and one DBP/E4BP4-binding element (D-box). Luciferase assays indicated that mTopoI gene expression was directly regulated by clock genes. The real-time monitoring assay showed that E-box and D-box response elements participate in the regulation of the circadian expression of mTopoI. Furthermore, a gel-shift assay showed that E2 is a direct target of the BMAL1/CLOCK heterodimer and DBP binds to the Putative D-site. These results indicate that TopoI is expressed in an autonomous circadian rhythm in NIH3T3 cells. (C) 2009 Elsevier Inc. All rights reserved.
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