title: Modified Nucleotides for Discrimination between Cytosine and the Epigenetic Marker 5-Methylcytosine.
abstract: 5-Methyl-2'-deoxycytosine, the most common epigenetic marker of DNA in eukaryotic cells, plays a key role in gene regulation and affects various cellular processes such as development and carcinogenesis. Therefore, the detection of 5mC can serve as an important biomarker for diagnostics. Here we describe that modified dGTP analogues as well as modified primers are able to sense the presence or absence of a single methylation of C, even though this modification does not interfere directly with Watson-Crick nucleobase pairing. By screening several modified nucleotide scaffolds, O(6)-modified 2'-deoxyguanosine analogues were identified as discriminating between C and 5mC. These modified nucleotides might find application in site-specific 5mC detection, for example, through real-time PCR approaches.
authors.affiliation: Fachbereich Chemie, Graduiertenschule Chemische, Biologie Konstanz, Universität Konstanz, Universitätsstrasse 10, 78457, Konstanz, Deutschland.
authors.forename: Janina
authors.initials: J
authors.lastname: von Watzdorf
authors.affiliation: Fachbereich Chemie, Graduiertenschule Chemische, Biologie Konstanz, Universität Konstanz, Universitätsstrasse 10, 78457, Konstanz, Deutschland.
authors.forename: Kim
authors.initials: K
authors.lastname: Leitner
authors.affiliation: Fachbereich Chemie, Graduiertenschule Chemische, Biologie Konstanz, Universität Konstanz, Universitätsstrasse 10, 78457, Konstanz, Deutschland. andreas.marx@uni-konstanz.de.
authors.forename: Andreas
authors.initials: A
authors.lastname: Marx
mesh_terms: D044503:5-Methylcytosine / Q000737:chemistry*; D003596:Cytosine / Q000737:chemistry*; D044127:Epigenesis, Genetic; D005819:Genetic Markers; D009711:Nucleotides / Q000737:chemistry*; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D005819:Genetic Markers; D009711:Nucleotides; D044503:5-Methylcytosine; D003596:Cytosine
keywords: 5-methylcytosine; DNA methylation; DNA polymerase; epigenetics; polymerase chain reaction
references.citation: J Am Chem Soc. 1968 Jan 17;90(2):474-8
references.pmid: 5634624
references.citation: Nat Rev Genet. 2010 Mar;11(3):204-20
references.pmid: 20142834
references.citation: Cell. 1992 Jun 12;69(6):915-26
references.pmid: 1606615
references.citation: Science. 2001 Aug 10;293(5532):1068-70
references.pmid: 11498573
references.citation: Methods. 2009 Mar;47(3):142-50
references.pmid: 18950712
references.citation: Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1827-31
references.pmid: 1542678
references.citation: Nat Genet. 1994 Mar;6(3):236-44
references.pmid: 8012384
references.citation: J Biol Chem. 1987 Oct 25;262(30):14689-96
references.pmid: 3667598
references.citation: Angew Chem Int Ed Engl. 2013 Apr 15;52(16):4350-5
references.pmid: 23559386
references.citation: Nat Rev Genet. 2012 Oct;13(10):679-92
references.pmid: 22945394
references.citation: Angew Chem Int Ed Engl. 2014 Jul 28;53(31):8154-8
references.pmid: 24923910
references.citation: Nat Protoc. 2006;1(5):2353-64
references.pmid: 17406479
references.citation: Angew Chem Int Ed Engl. 2016 Feb 24;55(9):3229-32
references.pmid: 26835661
references.citation: Nucleic Acids Res. 2008 Dec;36(22):e150
references.pmid: 18984622
references.citation: Proc Natl Acad Sci U S A. 1988 Sep;85(17):6252-6
references.pmid: 3413095
references.citation: Nature. 1984 Mar 8-14;308(5955):201-3
references.pmid: 6322009
references.citation: Chembiochem. 2013 Jun 17;14(9):1058-62
references.pmid: 23733496
references.citation: Science. 2012 May 11;336(6082):736-9
references.pmid: 22499810
references.citation: Nucleosides Nucleotides Nucleic Acids. 2009 May;28(5):408-23
references.pmid: 20183592
references.citation: Nat Rev Genet. 2002 Jun;3(6):415-28
references.pmid: 12042769
references.citation: Nature. 2010 Apr 15;464(7291):1082-6
references.pmid: 20393567
references.citation: J Am Chem Soc. 1970 Feb 11;92(3):724-6
references.pmid: 5411063
references.citation: Anal Biochem. 1998 Nov 1;264(1):129-32
references.pmid: 9784198
references.citation: Nature. 2008 Oct 23;455(7216):1124-8
references.pmid: 18806776
references.citation: Nucleic Acids Res. 2001 Jul 1;29(13):E65-5
references.pmid: 11433041
references.citation: J Am Chem Soc. 2013 Jan 30;135(4):1240-3
references.pmid: 23301810
references.citation: Angew Chem Int Ed Engl. 2008;47(1):181-4
references.pmid: 18041015
references.citation: Nat Rev Cancer. 2003 Apr;3(4):253-66
references.pmid: 12671664
references.citation: Nat Med. 2011 Mar;17(3):330-9
references.pmid: 21386836
references.citation: Genome Res. 2010 Oct;20(10):1369-82
references.pmid: 20716667
references.citation: Nucleic Acids Res. 1991 Aug 11;19(15):4045-57
references.pmid: 1870963
references.citation: PLoS One. 2013 Sep 04;8(9):e72435
references.pmid: 24023739
references.citation: J Biol Chem. 1992 Feb 5;267(4):2633-9
references.pmid: 1370828
references.citation: Proc Natl Acad Sci U S A. 1982 Dec;79(23):7440-4
references.pmid: 6961422
references.citation: Science. 2007 Dec 21;318(5858):1917-20
references.pmid: 18029452
references.citation: Nucleic Acids Res. 1979 Dec 20;7(8):2303-22
references.pmid: 523319