Yang, T;
Low, JJA;
Woon, ECY;
(2020)
A general strategy exploiting m⁵C duplex-remodelling effect for selective detection of RNA and DNA m⁵C methyltransferase activity in cells.
Nucleic Acids Research
, 48
(1)
, Article e5. 10.1093/nar/gkz1047.
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A general strategy exploiting m5C duplex-remodelling effect for selective detection of RNA and DNA m5C methyltransferase act.pdf - Accepted Version Download (6MB) | Preview |
Abstract
RNA:5-methylcytosine (m⁵C) methyltransferases are currently the focus of intense research following a series of high-profile reports documenting their physiological links to several diseases. However, no methods exist which permit the specific analysis of RNA:m⁵C methyltransferases in cells. Herein, we described how a combination of biophysical studies led us to identify distinct duplex-remodelling effects of m5C on RNA and DNA duplexes. Specifically, m⁵C induces a C3′-endo to C2′-endo sugar-pucker switch in CpG RNA duplex but triggers a B-to-Z transformation in CpG DNA duplex. Inspired by these different ‘structural signatures’, we developed a m⁵C-sensitive probe which fluoresces spontaneously in response to m5C-induced sugar-pucker switch, hence useful for sensing RNA:m⁵C methyltransferase activity. Through the use of this probe, we achieved real-time imaging and flow cytometry analysis of NOP2/Sun RNA methyltransferase 2 (NSUN2) activity in HeLa cells. We further applied the probe to the cell-based screening of NSUN2 inhibitors. The developed strategy could also be adapted for the detection of DNA:m⁵C methyltransferases. This was demonstrated by the development of DNA m⁵C-probe which permits the screening of DNA methyltransferase 3A inhibitors. To our knowledge, this study represents not only the first examples of m⁵C-responsive probes, but also a new strategy for discriminating RNA and DNA m⁵C methyltransferase activity in cells.
| Type: | Article |
|---|---|
| Title: | A general strategy exploiting m⁵C duplex-remodelling effect for selective detection of RNA and DNA m⁵C methyltransferase activity in cells |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1093/nar/gkz1047 |
| Publisher version: | https://doi.org/10.1093/nar/gkz1047 |
| Language: | English |
| Additional information: | © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/). |
| Keywords: | methyltransferase activity in cells. |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > UCL School of Pharmacy UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > UCL School of Pharmacy > Pharma and Bio Chemistry |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10106330 |
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