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Related Experiment Video

Updated: Jul 8, 2026

Sequence-specific Labeling of Nucleic Acids and Proteins with Methyltransferases and Cofactor Analogues

Sequence-specific Labeling of Nucleic Acids and Proteins with Methyltransferases and Cofactor Analogues

Published on: November 22, 2014

Methyltransferase-directed DNA strand scission.

Lindsay R Comstock¹, Scott R Rajski

¹School of Pharmacy, University of Wisconsin-Madison, 777 Highland Avenue, Madison, WI 53705-2222, USA.

Journal of the American Chemical Society

|October 13, 2005

Summary

This summary is machine-generated.

This study shows how modified DNA can be damaged to reveal methylation sites. This new chemical method rapidly identifies DNA methylation using specific DNA damage, aiding in genetic research.

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Continuous Fluorescence-Based Endonuclease-Coupled DNA Methylation Assay to Screen for DNA Methyltransferase Inhibitors

Continuous Fluorescence-Based Endonuclease-Coupled DNA Methylation Assay to Screen for DNA Methyltransferase Inhibitors

Published on: August 5, 2022

Area of Science:

Biochemistry
Organic Chemistry
Molecular Biology

Background:

DNA methylation is a crucial epigenetic modification involved in gene regulation.
Identifying methylated DNA regions is essential for understanding various biological processes and diseases.
Current methods for detecting DNA methylation can be complex and time-consuming.

Purpose of the Study:

To develop a novel chemical strategy for the rapid identification of DNA methylation.
To utilize Staudinger ligation chemistry for DNA modification and subsequent strand scission.
To establish a method for detecting DNA methyltransferase (MTase) activity and recognition sites.

Main Methods:

DNA modified by methyltransferases (MTases) was subjected to Staudinger ligation with phenanthroline-derivatized triarylphosphines.
The resulting duplexes were treated with Cu(II) and 3-mercaptopropionic acid to induce DNA strand scission.
Specific MTases (M.TaqI and M.HhaI) were used with synthetic azide-bearing cofactors to create DNA lesions.

Main Results:

MTase-modified DNA successfully underwent Staudinger ligation.
The chemical treatment led to strand scission specifically at the MTase recognition site.
DNA lesions were generated that induced strand scission 5' to the enzyme-modified base.

Conclusions:

A new chemical approach enables rapid identification of DNA methylation.
The method relies on inducing DNA damage proximal to methylation sites.
This technique offers a valuable tool for studying DNA methylation patterns and MTase activity.