ArticleNucleic acids research2010
A real-time assay for CpG-specific cytosine-C5 methyltransferase activity.
Article in Nucleic acids research, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 57 citations in OpenAlex.
- Phospho-Switch: Regulation of the Activity of SAM-Dependent Methyltransferases UsingInternational journal of molecular sciences · 2025Article
- Cytosine methylase and hydroxymethylase activity in mammalian mitochondria.Frontiers in cell and developmental biology · 2025Article
- Article
- A sequential one-pot approach for rapid and convenient characterization of putative restriction-modification systems.mSystems · 2023Article
- HpaII-assisted and linear amplification-enhanced isothermal exponential amplification fluorescent strategy for rapid and sensitive detection of DNA methyltransferase activity.Analytical and bioanalytical chemistry · 2023Article
- Z-DNA as a Tool for Nuclease-Free DNA Methyltransferase Assay.International journal of molecular sciences · 2021Article
- Article
- Structure-Guided Identification of DNMT3B Inhibitors.ACS medicinal chemistry letters · 2020Article
- An integrated-molecular-beacon based multiple exponential strand displacement amplification strategy for ultrasensitive detection of DNA methyltransferase activity.Chemical science · 2019Article
- Single-ribonucleotide repair-mediated ligation-dependent cycling signal amplification for sensitive and specific detection of DNA methyltransferase.Chemical science · 2018Article
- Inhibition of DNA Methylation at theACS omega · 2016Article
- Expression and purification of the modification-dependent restriction enzyme BisI and its homologous enzymes.Scientific reports · 2016Article
- DNA Methyltransferase Activity Assays: Advances and Challenges.Theranostics · 2016Review
- Electrochemical assay for the signal-on detection of human DNA methyltransferase activity.Journal of the American Chemical Society · 2013Article
- Laccaic acid A is a direct, DNA-competitive inhibitor of DNA methyltransferase 1.The Journal of biological chemistry · 2013Article
- Dissection of cell cycle-dependent dynamics of Dnmt1 by FRAP and diffusion-coupled modeling.Nucleic acids research · 2013Article
- The replication focus targeting sequence (RFTS) domain is a DNA-competitive inhibitor of Dnmt1.The Journal of biological chemistry · 2011Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A real-time assay for CpG-specific cytosine-C5 methyltransferase activity has been developed. The assay applies a break light oligonucleotide in which the methylation of an unmethylated 5'-CG-3' site is enzymatically coupled to the development of a fluorescent signal. This sensitive assay can measure rates of DNA methylation down to 0.34 +/- 0.06 fmol/s. The assay is reproducible, with a coefficient of variation over six independent measurements of 4.5%. Product concentration was accurately measured from fluorescence signals using a linear calibration curve, which achieved a goodness of fit (R(2)) above 0.98. The oligonucleotide substrate contains three C5-methylated cytosine residues and one unmethylated 5'-CG-3' site. Methylation yields an oligonucleotide containing the optimal substrate for the restriction enzyme GlaI. Cleavage of the fully methylated oligonucleotide leads to separation of fluorophore from quencher, giving a proportional increase in fluorescence. This method has been used to assay activity of DNMT1, the principle maintenance methyltransferase in human cells, and for the kinetic characterization of the bacterial cytosine-C5 methyltransferase M.SssI. The assay has been shown to be suitable for the real-time monitoring of DNMT1 activity in a high-throughput format, with low background signal and the ability to obtain linear rates of methylation over long periods, making this a promising method of high-throughput screening for inhibitors.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.