ArticleNucleic acids research2016
5-methylcytosine-sensitive variants of Thermococcus kodakaraensis DNA polymerase.
Article in Nucleic acids research, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 22 citations in OpenAlex.
- Semi-rational evolution of a recombinant DNA polymerase for modified nucleotide incorporation efficiency.PloS one · 2025Article
- Direct Enzyme Engineering of B Family DNA Polymerases for Biotechnological Approaches.Bioengineering (Basel, Switzerland) · 2023Review
- m5C methylation modification guides the prognostic value and immune landscapes in acute myeloid leukemia.Aging · 2023Article
- Cytosine base modifications regulate DNA duplex stability and metabolism.Nucleic acids research · 2021Article
- Review
- Evolution of a reverse transcriptase to map NNature methods · 2019Article
- Engineering Polymerases for New Functions.Trends in biotechnology · 2019Review
- Identification of Thermus aquaticus DNA polymerase variants with increased mismatch discrimination and reverse transcriptase activity from a smart enzyme mutant library.Scientific reports · 2019Article
- Engineering of a DNA Polymerase for Direct mAngewandte Chemie (International ed. in English) · 2018Article
- Engineering human PrimPol into an efficient RNA-dependent-DNA primase/polymerase.Nucleic acids research · 2017Article
- Exploring the Chemistry of Genetic Information Storage and Propagation through Polymerase Engineering.Accounts of chemical research · 2017Review
- Variants of sequence family B Thermococcus kodakaraensis DNA polymerase with increased mismatch extension selectivity.PloS one · 2017Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
DNA methylation of cytosine in eukaryotic cells is a common epigenetic modification, which plays an important role in gene expression and thus affects various cellular processes like development and carcinogenesis. The occurrence of 5-methyl-2'-deoxycytosine (5mC) as well as the distribution pattern of this epigenetic marker were shown to be crucial for gene regulation and can serve as important biomarkers for diagnostics. DNA polymerases distinguish little, if any, between incorporation opposite C and 5mC, which is not surprising since the site of methylation is not involved in Watson-Crick recognition. Here, we describe the development of a DNA polymerase variant that incorporates the canonical 2'-deoxyguanosine 5'-monophosphate (dGMP) opposite C with higher efficiency compared to 5mC. The variant of Thermococcus kodakaraensis (KOD) exo- DNA polymerase was discovered by screening mutant libraries that were built by rational design. We discovered that an amino acid substitution at a single site that does not directly interact with the templating nucleobase, may alter the ability of the DNA polymerase in processing C in comparison to 5mC. Employing these findings in combination with a nucleotide, which is fluorescently labeled at the terminal phosphate, indicates the potential use of the mutant DNA polymerase in the detection of 5mC.
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Registered trials
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