Content of 5me-Deoxycytidine (5-meC) in DNA preparations of ten species and the estimated average numbers of 5mdC per (haploid) genome

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Organism Various
Reference Capuano F, Mülleder M, Kok R, Blom HJ, Ralser M. Cytosine DNA methylation is found in Drosophila melanogaster but absent in Saccharomyces cerevisiae, Schizosaccharomyces pombe, and other yeast species. Anal Chem. 2014 Apr 15 86(8):3697-702. doi: 10.1021/ac500447w. p.3699 table 1PubMed ID24640988
Primary Source See refs beneath table
Method Abstract: "[Investigators] employed an alternative LC-MS/MS protocol where by targeting 5-methyldeoxycytidine moieties after enzymatic digestion, only 5-meC specifically derived from DNA is quantified. This technique unambiguously identified cytosine DNA methylation in Arabidopsis thaliana (14.0% of cytosines methylated), Mus musculus (7.6%), and Escherichia coli (2.3%)."
Comments P.3700 left column top paragraph: "Taking into account the genome size and GC content, these values allowed an estimation about the number of methylated cytosine residues per genome (A. thaliana, 6 × 10^6 M. musculus, 9 × 10^7 E. coli, 5 × 10^4) (Table 1). Moreover, this protocol confirmed the expected decline in methylation in the DNA methyltransferases deficient (dcm-6) K12 derivate GM2929 (Figure1b, Table 1). Compared to the methylation competent K12 strain DH5α, the content of methylated cytosines was reduced to 0.016% (equaling <500 modified cytosine bases per genome), which corresponds to a reduction in DNA methylation of 99.3%. Whereas, unmethylated cytidines were readily detected in all yeast species (Figure 1b, upper panel), the protocol applied on S. cerevisiae, S. boulardii, S. paradoxus, P. pastoris, and S. pombe DNA did not detect any evidence for the content of 5-methyldeoxycytidine (Figure ​1b, lower panel Table 1). Taking into account the limit of detection at 250 attomol for methyldeoxycytidine, its content in yeast would be lower than 0.00002 per deoxycytidine (Table 1)."
Entered by Uri M
ID 112844