Limits...
Formation and abundance of 5-hydroxymethylcytosine in RNA.

Huber SM, van Delft P, Mendil L, Bachman M, Smollett K, Werner F, Miska EA, Balasubramanian S - Chembiochem (2015)

Bottom Line: Herein, we describe an in vivo isotope-tracing methodology to demonstrate that the ribonucleoside 5-methylcytidine (m(5)C) is subject to oxidative processing in mammals, forming 5-hydroxymethylcytidine (hm(5)C) and 5-formylcytidine (f(5)C).Furthermore, we have identified hm(5)C in total RNA from all three domains of life and in polyA-enriched RNA fractions from mammalian cells.This suggests m(5)C oxidation is a conserved process that could have critical regulatory functions inside cells.

View Article: PubMed Central - PubMed

Affiliation: University of Cambridge, Department of Chemistry, Lensfield Road, Cambridge, CB2 1EW (UK).

No MeSH data available.


Synthesis of stable-isotope-labelled cytidine, 5-methylcytidine and 5-hydroxymethylcytidine: a) polyphosphoric acid, 17 h, 95 °C, 37 %; b) TMSCl, 1,1,1,3,3,3-hexamethyldisilazane, MeCN, 14 h, 85 °C; c) 1-O-acetyl-2,3,5-tri-O-benzoyl-β-D-ribofuranose, TMSOTf, 1,2-dichloroethene, 4 h, 70 °C, 80 %; d) tris(3,5-dihydro-4H-1,2,4-triazol-4-yl)phosphine oxide, MeCN, 17 h, 25 °C; e) NH4OH (35 %), 1,4-dioxane, 2 h, 25 °C, 69 %; f) NaOMe, MeOH, 5 h, 25 °C, 97 %; g) H2SO4, acetone, 15 h, 25 °C, 87 %; h) paraformaldehyde, KOH (0.5 M), microwave irradiation, 75 min, 60 °C, 16 %; i) 90 % TFA, 1 h, 0 °C, 98 %. Asterisks represent a 15N or 13C isotopologue.[18]
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4471624&req=5

fig04: Synthesis of stable-isotope-labelled cytidine, 5-methylcytidine and 5-hydroxymethylcytidine: a) polyphosphoric acid, 17 h, 95 °C, 37 %; b) TMSCl, 1,1,1,3,3,3-hexamethyldisilazane, MeCN, 14 h, 85 °C; c) 1-O-acetyl-2,3,5-tri-O-benzoyl-β-D-ribofuranose, TMSOTf, 1,2-dichloroethene, 4 h, 70 °C, 80 %; d) tris(3,5-dihydro-4H-1,2,4-triazol-4-yl)phosphine oxide, MeCN, 17 h, 25 °C; e) NH4OH (35 %), 1,4-dioxane, 2 h, 25 °C, 69 %; f) NaOMe, MeOH, 5 h, 25 °C, 97 %; g) H2SO4, acetone, 15 h, 25 °C, 87 %; h) paraformaldehyde, KOH (0.5 M), microwave irradiation, 75 min, 60 °C, 16 %; i) 90 % TFA, 1 h, 0 °C, 98 %. Asterisks represent a 15N or 13C isotopologue.[18]


Formation and abundance of 5-hydroxymethylcytosine in RNA.

Huber SM, van Delft P, Mendil L, Bachman M, Smollett K, Werner F, Miska EA, Balasubramanian S - Chembiochem (2015)

Synthesis of stable-isotope-labelled cytidine, 5-methylcytidine and 5-hydroxymethylcytidine: a) polyphosphoric acid, 17 h, 95 °C, 37 %; b) TMSCl, 1,1,1,3,3,3-hexamethyldisilazane, MeCN, 14 h, 85 °C; c) 1-O-acetyl-2,3,5-tri-O-benzoyl-β-D-ribofuranose, TMSOTf, 1,2-dichloroethene, 4 h, 70 °C, 80 %; d) tris(3,5-dihydro-4H-1,2,4-triazol-4-yl)phosphine oxide, MeCN, 17 h, 25 °C; e) NH4OH (35 %), 1,4-dioxane, 2 h, 25 °C, 69 %; f) NaOMe, MeOH, 5 h, 25 °C, 97 %; g) H2SO4, acetone, 15 h, 25 °C, 87 %; h) paraformaldehyde, KOH (0.5 M), microwave irradiation, 75 min, 60 °C, 16 %; i) 90 % TFA, 1 h, 0 °C, 98 %. Asterisks represent a 15N or 13C isotopologue.[18]
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4471624&req=5

fig04: Synthesis of stable-isotope-labelled cytidine, 5-methylcytidine and 5-hydroxymethylcytidine: a) polyphosphoric acid, 17 h, 95 °C, 37 %; b) TMSCl, 1,1,1,3,3,3-hexamethyldisilazane, MeCN, 14 h, 85 °C; c) 1-O-acetyl-2,3,5-tri-O-benzoyl-β-D-ribofuranose, TMSOTf, 1,2-dichloroethene, 4 h, 70 °C, 80 %; d) tris(3,5-dihydro-4H-1,2,4-triazol-4-yl)phosphine oxide, MeCN, 17 h, 25 °C; e) NH4OH (35 %), 1,4-dioxane, 2 h, 25 °C, 69 %; f) NaOMe, MeOH, 5 h, 25 °C, 97 %; g) H2SO4, acetone, 15 h, 25 °C, 87 %; h) paraformaldehyde, KOH (0.5 M), microwave irradiation, 75 min, 60 °C, 16 %; i) 90 % TFA, 1 h, 0 °C, 98 %. Asterisks represent a 15N or 13C isotopologue.[18]
Bottom Line: Herein, we describe an in vivo isotope-tracing methodology to demonstrate that the ribonucleoside 5-methylcytidine (m(5)C) is subject to oxidative processing in mammals, forming 5-hydroxymethylcytidine (hm(5)C) and 5-formylcytidine (f(5)C).Furthermore, we have identified hm(5)C in total RNA from all three domains of life and in polyA-enriched RNA fractions from mammalian cells.This suggests m(5)C oxidation is a conserved process that could have critical regulatory functions inside cells.

View Article: PubMed Central - PubMed

Affiliation: University of Cambridge, Department of Chemistry, Lensfield Road, Cambridge, CB2 1EW (UK).

No MeSH data available.