Journal
FEBS JOURNAL
Volume 277, Issue 20, Pages 4254-4264Publisher
WILEY
DOI: 10.1111/j.1742-4658.2010.07814.x
Keywords
G-quadruplex; NMR; thymidine kinase 1
Categories
Funding
- CSIR
- Department of Science and Technology [DST/SJF/LS-03]
- CSIR Task Force Project [CMM 0017]
- Singapore Ministry of Education [ARC30/07]
- Nanyang Technological University (NTU) [RG62/07]
- Singapore Biomedical Research Council [07/1/22/19/542]
- Division of Chemistry and Biological Chemistry
- NTU School of Biological Sciences
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G-quadruplex motifs constitute unusual DNA secondary structures formed by stacking of planar hydrogen-bonded G-tetrads. Recent genome-wide bioinformatics and experimental analyses have suggested the interesting possibility that G-quadruplex motifs could be cis-regulatory elements. Here, we identified a characteristic potential G-quadruplex-forming sequence element within the promoter of human thymidine kinase 1 (TK1). Our NMR, UV and CD spectroscopy and gel electrophoresis data suggested that this sequence forms a novel intramolecular G-quadruplex with two G-tetrads in K+ solution. The results presented here indicate the role of this G-quadruplex motif in transcription of TK1 in cell-based reporter assays. Specific nucleotide substitutions designed to destabilize the G-quadruplex motif resulted in increased promoter activity, supporting direct involvement of the G-quadruplex motif in transcription of TK1. These studies suggest that the G-quadruplex motif may be an important target for controlling critical biological processes, such as DNA synthesis, mediated by TK1.
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