Ascorbic acid enhances tet-mediated 5-methylcytosine oxidation and promotes DNA demethylation in mammals

Ruichuan Yin, Shi Qing Mao, Bailin Zhao, Zechen Chong, Ying Yang, Chao Zhao, Dapeng Zhang, Hua Huang, Juan Gao, Zheng Li, Yan Jiao, Cuiping Li, Shengquan Liu, Danni Wu, Weikuan Gu, Yun Gui Yang, Guo Liang Xu, Hailin Wang

Research output: Contribution to journalArticle

233 Citations (Scopus)

Abstract

DNA hydroxymethylation and its mediated DNA demethylation are critical for multiple cellular processes, for example, nuclear reprogramming, embryonic development, and many diseases. Here, we demonstrate that a vital nutrient ascorbic acid (AA), or vitamin C (Vc), can directly enhance the catalytic activity of Tet dioxygenases for the oxidation of 5-methylcytosine (5mC). As evidenced by changes in intrinsic fluorescence and catalytic activity of Tet2 protein caused by AA and its oxidation-resistant derivatives, we further show that AA can uniquely interact with the C-terminal catalytic domain of Tet enzymes, which probably promotes their folding and/or recycling of the cofactor Fe2+. Other strong reducing chemicals do not have a similar effect. These results suggest that AA also acts as a cofactor of Tet enzymes. In mouse embryonic stem cells, AA significantly increases the levels of all 5mC oxidation products, particularly 5-formylcytosine and 5-carboxylcytosine (by more than an order of magnitude), leading to a global loss of 5mC (∼40%). In cells deleted of the Tet1 and Tet2 genes, AA alters neither 5mC oxidation nor the overall level of 5mC. The AA effects are however restored when Tet2 is re-expressed in the Tet-deficient cells. The enhancing effects of AA on 5mC oxidation and DNA demethylation are also observed in a mouse model deficient in AA synthesis. Our data establish a direct link among AA, Tet, and DNA methylation, thus revealing a role of AA in the regulation of DNA modifications.

Original languageEnglish (US)
Pages (from-to)10396-10403
Number of pages8
JournalJournal of the American Chemical Society
Volume135
Issue number28
DOIs
StatePublished - Jul 17 2013

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5-Methylcytosine
Mammals
Ascorbic acid
Ascorbic Acid
DNA
Oxidation
Catalyst activity
Enzymes
Dioxygenases
Vitamins
Coenzymes
Stem cells
Recycling
DNA Methylation
Nutrients
Embryonic Development
Catalytic Domain
Genes

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Ascorbic acid enhances tet-mediated 5-methylcytosine oxidation and promotes DNA demethylation in mammals. / Yin, Ruichuan; Mao, Shi Qing; Zhao, Bailin; Chong, Zechen; Yang, Ying; Zhao, Chao; Zhang, Dapeng; Huang, Hua; Gao, Juan; Li, Zheng; Jiao, Yan; Li, Cuiping; Liu, Shengquan; Wu, Danni; Gu, Weikuan; Yang, Yun Gui; Xu, Guo Liang; Wang, Hailin.

In: Journal of the American Chemical Society, Vol. 135, No. 28, 17.07.2013, p. 10396-10403.

Research output: Contribution to journalArticle

Yin, R, Mao, SQ, Zhao, B, Chong, Z, Yang, Y, Zhao, C, Zhang, D, Huang, H, Gao, J, Li, Z, Jiao, Y, Li, C, Liu, S, Wu, D, Gu, W, Yang, YG, Xu, GL & Wang, H 2013, 'Ascorbic acid enhances tet-mediated 5-methylcytosine oxidation and promotes DNA demethylation in mammals', Journal of the American Chemical Society, vol. 135, no. 28, pp. 10396-10403. https://doi.org/10.1021/ja4028346
Yin, Ruichuan ; Mao, Shi Qing ; Zhao, Bailin ; Chong, Zechen ; Yang, Ying ; Zhao, Chao ; Zhang, Dapeng ; Huang, Hua ; Gao, Juan ; Li, Zheng ; Jiao, Yan ; Li, Cuiping ; Liu, Shengquan ; Wu, Danni ; Gu, Weikuan ; Yang, Yun Gui ; Xu, Guo Liang ; Wang, Hailin. / Ascorbic acid enhances tet-mediated 5-methylcytosine oxidation and promotes DNA demethylation in mammals. In: Journal of the American Chemical Society. 2013 ; Vol. 135, No. 28. pp. 10396-10403.
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AU - Yin, Ruichuan

AU - Mao, Shi Qing

AU - Zhao, Bailin

AU - Chong, Zechen

AU - Yang, Ying

AU - Zhao, Chao

AU - Zhang, Dapeng

AU - Huang, Hua

AU - Gao, Juan

AU - Li, Zheng

AU - Jiao, Yan

AU - Li, Cuiping

AU - Liu, Shengquan

AU - Wu, Danni

AU - Gu, Weikuan

AU - Yang, Yun Gui

AU - Xu, Guo Liang

AU - Wang, Hailin

PY - 2013/7/17

Y1 - 2013/7/17

N2 - DNA hydroxymethylation and its mediated DNA demethylation are critical for multiple cellular processes, for example, nuclear reprogramming, embryonic development, and many diseases. Here, we demonstrate that a vital nutrient ascorbic acid (AA), or vitamin C (Vc), can directly enhance the catalytic activity of Tet dioxygenases for the oxidation of 5-methylcytosine (5mC). As evidenced by changes in intrinsic fluorescence and catalytic activity of Tet2 protein caused by AA and its oxidation-resistant derivatives, we further show that AA can uniquely interact with the C-terminal catalytic domain of Tet enzymes, which probably promotes their folding and/or recycling of the cofactor Fe2+. Other strong reducing chemicals do not have a similar effect. These results suggest that AA also acts as a cofactor of Tet enzymes. In mouse embryonic stem cells, AA significantly increases the levels of all 5mC oxidation products, particularly 5-formylcytosine and 5-carboxylcytosine (by more than an order of magnitude), leading to a global loss of 5mC (∼40%). In cells deleted of the Tet1 and Tet2 genes, AA alters neither 5mC oxidation nor the overall level of 5mC. The AA effects are however restored when Tet2 is re-expressed in the Tet-deficient cells. The enhancing effects of AA on 5mC oxidation and DNA demethylation are also observed in a mouse model deficient in AA synthesis. Our data establish a direct link among AA, Tet, and DNA methylation, thus revealing a role of AA in the regulation of DNA modifications.

AB - DNA hydroxymethylation and its mediated DNA demethylation are critical for multiple cellular processes, for example, nuclear reprogramming, embryonic development, and many diseases. Here, we demonstrate that a vital nutrient ascorbic acid (AA), or vitamin C (Vc), can directly enhance the catalytic activity of Tet dioxygenases for the oxidation of 5-methylcytosine (5mC). As evidenced by changes in intrinsic fluorescence and catalytic activity of Tet2 protein caused by AA and its oxidation-resistant derivatives, we further show that AA can uniquely interact with the C-terminal catalytic domain of Tet enzymes, which probably promotes their folding and/or recycling of the cofactor Fe2+. Other strong reducing chemicals do not have a similar effect. These results suggest that AA also acts as a cofactor of Tet enzymes. In mouse embryonic stem cells, AA significantly increases the levels of all 5mC oxidation products, particularly 5-formylcytosine and 5-carboxylcytosine (by more than an order of magnitude), leading to a global loss of 5mC (∼40%). In cells deleted of the Tet1 and Tet2 genes, AA alters neither 5mC oxidation nor the overall level of 5mC. The AA effects are however restored when Tet2 is re-expressed in the Tet-deficient cells. The enhancing effects of AA on 5mC oxidation and DNA demethylation are also observed in a mouse model deficient in AA synthesis. Our data establish a direct link among AA, Tet, and DNA methylation, thus revealing a role of AA in the regulation of DNA modifications.

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