The discovery of 5mC-selective deaminases and their application to ultra-sensitive direct sequencing of methylated sites at base resolution.
Yang, W.; Lee, Y.-J.; Silva, R. M. B.; DeLiberto, A.; Yancey, C.; McCallum, D.; Buss, J.; Moncion, R.; Ong, J.; Mabuchi, M.; Hough, D.; Weigele, P. R.; Ettwiller, L. M.
Show abstract
Mining phages for new enzymatic activities continues to be important for the development of new tools for biotechnology. In this study, we used MetaGPA--a method linking genotype to phenotype in metagenomic data--to identify deoxycytidine deaminases, a protein family highly associated with cytosine modifications in metaviromes. Unexpectedly, a subset of these deaminases exhibited a preference for 5-methylcytosine (5mC) over cytosine (C) in both mononucleotide and single-stranded DNA substrates. In a methylome sequencing workflow, preferential deamination of 5mC by these enzymes enabled direct conversion of methylated cytosine while completely eliminating any background deamination of unmodified cytosine. This direct conversion allows for precise identification of methylated sites at single-base resolution with unmatched sensitivity enabling broad applications for the simultaneous sequencing of genome and methylome.
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