Formation and Abundance of 5-Hydroxymethylcytosine in RNA

• Published in: Chembiochem : a European journal of chemical biology Vol. 16; no. 5; pp. 752 - 755
• Main Authors: Huber, Sabrina M., van Delft, Pieter, Mendil, Lee, Bachman, Martin, Smollett, Katherine, Werner, Finn, Miska, Eric A., Balasubramanian, Shankar
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 23.03.2015; WILEY‐VCH Verlag; Wiley; Wiley Subscription Services, Inc
• Subjects: 5-hydroxymethylcytosine; 5-methylcytosine; 5-Methylcytosine - analogs & derivatives; Animals; Biochemistry & Molecular Biology; Chemistry, Medicinal; Chromatography, High Pressure Liquid; Communications; Cytosine - analogs & derivatives; Cytosine - biosynthesis; Cytosine - chemistry; Cytosine - metabolism; isotope tracing; LC-MS/MS; Life Sciences & Biomedicine; Mammals; Mice; Mice, Inbred C57BL; Molecular Structure; Oxidation-Reduction; Pharmacology & Pharmacy; RNA - chemistry; RNA - metabolism; RNA modifications; Science & Technology; Tandem Mass Spectrometry; NONCODING RNA; LC-MS; METHYLATION; MS; DNA; NUCLEOSIDES; 5-hydroxymethylcytosine; isotope tracing; 5-methylcytosine; RNA modifications; LC-MS/MS
• ISSN: 1439-4227; 1439-7633; 1439-7633
• DOI: 10.1002/cbic.201500013

RNA methylation is emerging as a regulatory RNA modification that could have important roles in the control and coordination of gene transcription and protein translation. Herein, we describe an in vivo isotope‐tracing methodology to demonstrate that the ribonucleoside 5‐methylcytidine (m5C) is subject to oxidative processing in mammals, forming 5‐hydroxymethylcytidine (hm5C) and 5‐formylcytidine (f5C). Furthermore, we have identified hm5C in total RNA from all three domains of life and in polyA‐enriched RNA fractions from mammalian cells. This suggests m5C oxidation is a conserved process that could have critical regulatory functions inside cells. Oxidative metabolites of 5‐methylcytosine: We show for the first time that 5‐hydroxymethylcytosine (hm5C) in RNA is generated by m5C oxidation in vivo and is pervasive in all three domains of life across different species. Our findings suggest that m5C oxidation is a conserved process that could have critical regulatory functions inside cells.