Abstract
Guanine-rich nucleic acid sequences can adopt noncanonical four-stranded secondary structures called guanine (G)-quadruplexes1. Bioinformatics analysis suggests that G-quadruplex motifs are prevalent in genomes2, which raises the need to elucidate their function. There is now evidence for the existence of DNA G-quadruplexes at telomeres with associated biological function3. A recent hypothesis supports the notion that gene promoter elements contain DNA G-quadruplex motifs that control gene expression at the transcriptional level4. We discovered a highly conserved, thermodynamically stable RNA G-quadruplex in the 5′ untranslated region (UTR) of the gene transcript of the human NRAS proto-oncogene. Using a cell-free translation system coupled to a reporter gene assay, we have demonstrated that this NRAS RNA G-quadruplex modulates translation. This is the first example of translational repression by an RNA G-quadruplex. Bioinformatics analysis has revealed 2,922 other 5′ UTR RNA G-quadruplex elements in the human genome. We propose that RNA G-quadruplexes in the 5′ UTR modulate gene expression at the translational level.
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Acknowledgements
We thank Cancer Research UK, the Cambridge Commonwealth Trust and Trinity College, Cambridge for funding. S.B. is a Biotechnology and Biological Sciences Research Council career development fellow. J.L.H. is a research fellow at Trinity College, Cambridge. We thank Z. Jawad-Alami for useful discussions and S. Sewitz for critically reading this manuscript.
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S.K., A.B. and S.B. conceived and designed the experiments. S.K. performed all the experiments. S.K. and A.B. analyzed the experimental data. J.L.H. conceived, designed and carried out the bioinformatics studies. All four authors discussed the results, and the manuscript was written by S.K. and S.B. with contributions from A.B. and J.L.H.
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Supplementary information
Supplementary Fig. 1
cDNA sequence of NRAS 5′ UTR. (PDF 98 kb)
Supplementary Fig. 2
CD melting of NRQ RNA G quadruplex. (PDF 37 kb)
Supplementary Table 1
Second G-quadruplex sequence motif in the NRAS 5′ UTRs of mouse and dog. (PDF 66 kb)
Supplementary Table 2
UV-melting temperatures of NRQ RNA G quadruplex at various strand concentrations. (PDF 18 kb)
Supplementary Table 3
UV-melting temperatures of NRQ RNA G quadruplex with different monovalent cations. (PDF 19 kb)
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Kumari, S., Bugaut, A., Huppert, J. et al. An RNA G-quadruplex in the 5′ UTR of the NRAS proto-oncogene modulates translation. Nat Chem Biol 3, 218–221 (2007). https://doi.org/10.1038/nchembio864
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DOI: https://doi.org/10.1038/nchembio864
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