ArticleNucleic acids research2010
A one-step method for quantitative determination of uracil in DNA by real-time PCR.
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 19 papers.
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Who cites it
19 citing papers in PubMed, 43 citations in OpenAlex.
- Ovulation sources ROS to confer mutagenic activities on the TP53 gene in the fallopian tube epithelium.Neoplasia (New York, N.Y.) · 2025Article
- Cytidine deaminases APOBEC3C and APOBEC3D promote DNA replication stress resistance in pancreatic cancer cells.Nature cancer · 2024Article
- Base-Resolution Analysis of Deoxyuridine at the Genome Scale Based on the Artificial Incorporation Modified Nucleobase.ACS central science · 2021Article
- Detection of Genomic Uracil Patterns.International journal of molecular sciences · 2021Review
- Visualization of uracils created by APOBEC3A using UdgX shows colocalization with RPA at stalled replication forks.Nucleic acids research · 2020Article
- Genome-wide alterations of uracil distribution patterns in human DNA upon chemotherapeutic treatments.eLife · 2020Article
- DCTPP1 prevents a mutator phenotype through the modulation of dCTP, dTTP and dUTP pools.Cellular and molecular life sciences : CMLS · 2020Article
- The etiology of uracil residues in the Saccharomyces cerevisiae genomic DNA.Current genetics · 2019Review
- Article
- Genome-wide mapping reveals that deoxyuridine is enriched in the human centromeric DNA.Nature chemical biology · 2018Article
- Differential control of dNTP biosynthesis and genome integrity maintenance by the dUTPase superfamily enzymes.Scientific reports · 2017Article
- The cell cycle restricts activation-induced cytidine deaminase activity to early G1.The Journal of experimental medicine · 2017Article
- Detection of uracil within DNA using a sensitive labeling method for in vitro and cellular applications.Nucleic acids research · 2016Article
- SHMT1 knockdown induces apoptosis in lung cancer cells by causing uracil misincorporation.Cell death & disease · 2014Article
- Methylated DNA is over-represented in whole-genome bisulfite sequencing data.Frontiers in genetics · 2014Article
- Uracil-containing DNA in Drosophila: stability, stage-specific accumulation, and developmental involvement.PLoS genetics · 2012Article
- The dUTPase enzyme is essential in Mycobacterium smegmatis.PloS one · 2012Article
- HIV DNA is heavily uracilated, which protects it from autointegration.Proceedings of the National Academy of Sciences of the United States of America · 2011Article
- Calpain-catalyzed proteolysis of human dUTPase specifically removes the nuclear localization signal peptide.PloS one · 2011Article
Corrections and comments
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Authors and funding
2 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Uracil may occur in DNA due to either cytosine deamination or thymine replacing incorporation. Its quantitative characterization is important in assessing DNA damages in cells with perturbed thymidylate metabolism or within different DNA segments involved in immunoglobulin gene diversification. The archaeal DNA polymerase from Pyrococcus furiosus binds strongly to the deaminated base uracil and stalls on uracil-containing templates. Here, we present a straightforward method for quantitative assessment of uracil in DNA within specific genomic segments. We use wild-type P. furiosus polymerase in parallel with its point mutant version which lacks the uracil-binding specificity on synthetic and genomic DNA samples to quantify the uracil content in a single-step real-time PCR assay. Quantification of the PCR results is based on an approach analogous to template copy number determination in comparing different samples. Data obtained on synthetic uracil-containing templates are verified by direct isotopic measurements. The method is also tested on physiological DNA samples from Escherichia coli and mouse cell lines with perturbed thymidylate biosynthesis. The present PCR-based method is easy to use and measures the uracil content within a genomic segment defined by the primers. Using distinct sets of primers, the method allows the analysis of heterogeneity of uracil distribution within the genome.
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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.