5mC oxidation products in the nucleic acids of the Basidiomycete fungi Laccaria bicolor and Coprinopsis cinerea

We analyzed oxidized 5-methylcytosine derivatives 5-hydroxymethylcytosine, 5-formylcytosine and 5-carboxylcytosine in nucleic acids of multicellular fungi Laccaria bicolor and Coprinopsis cinerea which have been used as models to study DNA methylation, developmental processes and symbiotic interactions. All three cytosine derivatives were detected in the genomes of both fungi, and importantly, we discovered 5carC in the RNA fractions, potentially including large non-coding, messenger RNAs and small RNA molecules, indicating gene regulatory functions of 5carC. Overall design: 5-hydroxymethylcytosine and 5-formylcytosine analysis in Basidiomycete fungi Laccaria bicolor and Coprinopsis cinerea using hMe-Seal, fC-Seal, hmTOP-seq and foTOP-seq methods. Vitamin C was used to increace amounts of 5-hydroxymethylcytosine and 5-formylcytosine in the hmTOP-seq and foTOP-seq methods for L.bicolor