Application of 5-Methylcytosine DNA Glycosylase to the Quantitative Analysis of DNA Methylation

Autor: Young Geun Mok, Woo Lee Choi, Jin Hoe Huh
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Methyltransferase
Arabidopsis
DEMETER
Polymerase Chain Reaction
Catalysis
Article
DNA Glycosylases
Epigenesis
Genetic
Inorganic Chemistry
lcsh:Chemistry
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
Solanum lycopersicum
Gene Expression Regulation
Plant
Epigenetics
Physical and Theoretical Chemistry
Molecular Biology
epiallele
lcsh:QH301-705.5
Spectroscopy
Alleles
030304 developmental biology
Homeodomain Proteins
0303 health sciences
DNA methylation
Chemistry
Arabidopsis Proteins
Organic Chemistry
General Medicine
DNA
Sequence Analysis
DNA
Computer Science Applications
Chromatin
DNA Demethylation
genomic DNA
DNA demethylation
Phenotype
Biochemistry
lcsh:Biology (General)
lcsh:QD1-999
DNA glycosylase
DNA demethylase
030220 oncology & carcinogenesis
Mutation
epigenetic profiling
Transcription Factors
Zdroj: International Journal of Molecular Sciences, Vol 22, Iss 1072, p 1072 (2021)
International Journal of Molecular Sciences
Volume 22
Issue 3
ISSN: 1661-6596
1422-0067
Popis: In higher eukaryotes DNA methylation is a prominent epigenetic mark important for chromatin structure and gene expression. Thus, profiling DNA methylation is important for predicting gene expressions associated with specific traits or diseases. DNA methylation is achieved by DNA methyltransferases and can be actively removed by specific enzymes in a replication-independent manner. DEMETER (DME) is a bifunctional 5-methylcytosine (5mC) DNA glycosylase responsible for active DNA demethylation that excises 5mC from DNA and cleaves a sugar-phosphate bond generating a single strand break (SSB). In this study, DME was used to analyze DNA methylation levels at specific epialleles accompanied with gain or loss of DNA methylation. DME treatment on genomic DNA generates SSBs in a nonsequence-specific fashion proportional to 5mC density, and thus DNA methylation levels can be easily measured when combined with the quantitative PCR (qPCR) method. The DME-qPCR analysis was applied to measure DNA methylation levels at the FWA gene in late-flowering Arabidopsis mutants and the CNR gene during fruit ripening in tomato. Differentially methylated epialleles were successfully distinguished corresponding to their expression levels and phenotypes. DME-qPCR is proven a simple yet effective method for quantitative DNA methylation analysis, providing advantages over current techniques based on methylation-sensitive restriction digestion.
Databáze: OpenAIRE
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