N6-Hydroperoxymethyladenosine: a new intermediate of chemical oxidation of N6-methyladenosine mediated by bicarbonate-activated hydrogen peroxide

Jinjun Wu; Heng Xiao; Tianlu Wang; Tingting Hong; Boshi Fu; Dongsheng Bai; Zhiyong He; Shuang Peng; Xiwen Xing; Jianlin Hu; Pu Guo; Xiang Zhou

doi:10.1039/c5sc00484e

N⁶-Hydroperoxymethyladenosine: a new intermediate of chemical oxidation of N⁶-methyladenosine mediated by bicarbonate-activated hydrogen peroxide

Chem Sci. 2015 May 1;6(5):3013-3017. doi: 10.1039/c5sc00484e. Epub 2015 Mar 11.

Authors

Jinjun Wu^{1

2}, Heng Xiao^{1

2}, Tianlu Wang^{1

2}, Tingting Hong^{1

2}, Boshi Fu^{1

2}, Dongsheng Bai^{1

2}, Zhiyong He^{1

2}, Shuang Peng^{1

2}, Xiwen Xing^{1

2}, Jianlin Hu^{1

2}, Pu Guo^{1

2}, Xiang Zhou^{1

2}

Affiliations

¹ College of Chemistry and Molecular Sciences , Institute of Advanced Studies , Wuhan University , Wuhan , Hubei 430072 , P. R. China . Email: xzhou@whu.edu.cn ; ; Tel: +86-27-68756663.
² State Key Laboratory of Natural and Biomimetic Drugs , Peking University , Beijing , 100191 , P. R. China.

Abstract

N⁶-Methyladenosine (m⁶A) represents a relatively abundant modification in eukaryotic RNA. Because m⁶A has similar properties to adenosine and a low reactivity, limited research has been focused on this nucleoside. In this study, we revealed an important intermediate in the oxidation of m⁶A through the bicarbonate-activated peroxide system. Over the course of oxidation, we found a new mechanism in which N⁶-hydroxymethyladenosine (hm⁶A), N⁶-formyladenosine (f⁶A) and N⁶-hydroperoxymethyladenosine (oxm⁶A) were intermediate products, and adenosine was the final product. In this study, oxm⁶A was isolated using HPLC and characterized by mass spectrometry, NMR and diphenyl-1-pyrenylphosphine (DPPP) fluorescence detection. This study provides a new modified nucleoside and demonstrates oxidative demethylation of m⁶A by reactive oxygen species at the nucleobase level and in RNA strands.