title: Quantification of 5-methyl-2'-deoxycytidine in the DNA.
abstract: Methylation at position 5 of cytosine (Cyt) at the CpG sequences leading to formation of 5-methyl-cytosine (m(5)Cyt) is an important element of epigenetic regulation of gene expression. Modification of the normal methylation pattern, unique to each organism, leads to the development of pathological processes and diseases, including cancer. Therefore, quantification of the DNA methylation and analysis of changes in the methylation pattern is very important from a practical point of view and can be used for diagnostic purposes, as well as monitoring of the treatment progress. In this paper we present a new method for quantification of 5-methyl-2'deoxycytidine (m(5)C) in the DNA. The technique is based on conversion of m(5)C into fluorescent 3,N(4)-etheno-5-methyl-2'deoxycytidine (εm(5)C) and its identification by reversed-phase high-performance liquid chromatography (RP-HPLC). The assay was used to evaluate m(5)C concentration in DNA of calf thymus and peripheral blood of cows bred under different conditions. This approach can be applied for measuring of 5-methylcytosine in cellular DNA from different cells and tissues.
authors.affiliation: Institute of Bioorganic Chemistry, Polish Academy of Sciences, Poznań, Poland.
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Giel-Pietraszuk
authors.affiliation: Faculty of Chemistry, Adam Mickiewicz University in Poznan, Poznań, Poland.
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Insińska-Rak
authors.affiliation: Faculty of Chemistry, Adam Mickiewicz University in Poznan, Poznań, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Golczak
authors.affiliation: Faculty of Chemistry, Adam Mickiewicz University in Poznan, Poznań, Poland.
authors.forename: Marek
authors.initials: M
authors.lastname: Sikorski
authors.affiliation: Institute of Bioorganic Chemistry, Polish Academy of Sciences, Poznań, Poland.
authors.forename: Mirosława
authors.initials: M
authors.lastname: Barciszewska
authors.affiliation: Institute of Bioorganic Chemistry, Polish Academy of Sciences, Poznań, Poland.
authors.forename: Jan
authors.initials: J
authors.lastname: Barciszewski
mesh_terms: D000818:Animals; D002138:Calibration; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase; D004247:DNA / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D019175:DNA Methylation; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000032:analysis / Q000097:blood / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D005260:Female; D057230:Limit of Detection; D008967:Molecular Biology / Q000379:methods*; D010761:Phosphorus Radioisotopes / Q000737:chemistry
chemical_list: D010761:Phosphorus Radioisotopes; D003841:Deoxycytidine; C032222:3,N(4)-ethenodeoxycytidine; D004247:DNA; C427203:calf thymus DNA; C016569:5-methyldeoxycytidine