Crosslinking of (Cytosine-5)-DNA Methyltransferase SsoII and its Complexes with Specific DNA Duplexes Provides an Insight into Their Structures

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.

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2D1424 Structural insight into maintenance methylation by mouse DNA methyltransferase 1(Dnmt1)(Protein: Structure & Function 1,The 48th Annual Meeting of the Biophysical Society of Japan)

Seibutsu Butsuri ◽

10.2142/biophys.51.s79_5 ◽

2011 ◽

Vol 51 (supplement) ◽

pp. S79-S80

Author(s):

Kohei Takeshita ◽

Isao Suetake ◽

Eiki Yamashita ◽

Michihiro Suga ◽

Hirotaka Narita ◽

...

Keyword(s):

Protein Structure ◽

Structure Function ◽

Annual Meeting ◽

Dna Methyltransferase ◽

Dna Methyltransferase 1 ◽

Biophysical Society ◽

Structural Insight ◽

Insight Into ◽

Maintenance Methylation ◽

Methyltransferase 1

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Insight into the selective binding mechanism of DNMT1 and DNMT3A inhibitors: a molecular simulation study

Physical Chemistry Chemical Physics ◽

10.1039/c9cp02024a ◽

2019 ◽

Vol 21 (24) ◽

pp. 12931-12947 ◽

Cited By ~ 10

Author(s):

Tianli Xie ◽

Jie Yu ◽

Weitao Fu ◽

Zhe Wang ◽

Lei Xu ◽

...

Keyword(s):

Molecular Simulation ◽

Simulation Study ◽

Dna Methyltransferase ◽

Rational Design ◽

Binding Mechanism ◽

Selective Inhibitors ◽

Selective Binding ◽

Simulation Techniques ◽

Insight Into

Molecular simulation techniques help with the rational design of novel selective inhibitors targeting certain DNA methyltransferase isoforms, which is beneficial for more refined treatments of epigenetic related cancer and other diseases.

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