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. 2015 May:82:22-8.
doi: 10.1016/j.freeradbiomed.2015.01.028. Epub 2015 Feb 3.

Oxidative stress alters global histone modification and DNA methylation

Yingmei Niu  1 , Thomas L DesMarais  2 , Zhaohui Tong  3 , Yixin Yao  4 , Max Costa  5
Affiliations

Oxidative stress alters global histone modification and DNA methylation

Yingmei Niu et al. Free Radic Biol Med. 2015 May.

Abstract

The JmjC domain-containing histone demethylases can remove histone lysine methylation and thereby regulate gene expression. The JmjC domain uses iron Fe(II) and α-ketoglutarate (αKG) as cofactors in an oxidative demethylation reaction via hydroxymethyl lysine. We hypothesize that reactive oxygen species will oxidize Fe(II) to Fe(III), thereby attenuating the activity of JmjC domain-containing histone demethylases. To minimize secondary responses from cells, extremely short periods of oxidative stress (3h) were used to investigate this question. Cells that were exposed to hydrogen peroxide (H2O2) for 3h exhibited increases in several histone methylation marks including H3K4me3 and decreases of histone acetylation marks including H3K9ac and H4K8ac; preincubation with ascorbate attenuated these changes. The oxidative stress level was measured by generation of 2',7'-dichlorofluorescein, GSH/GSSG ratio, and protein carbonyl content. A cell-free system indicated that H2O2 inhibited histone demethylase activity where increased Fe(II) rescued this inhibition. TET protein showed a decreased activity under oxidative stress. Cells exposed to a low-dose and long-term (3 weeks) oxidative stress also showed increased global levels of H3K4me3 and H3K27me3. However, these global methylation changes did not persist after washout. The cells exposed to short-term oxidative stress also appeared to have higher activity of class I/II histone deacetylase (HDAC) but not class III HDAC. In conclusion, we have found that oxidative stress transiently alters the epigenetic program process through modulating the activity of enzymes responsible for demethylation and deacetylation of histones.

Keywords: Histone demethylase; Hydrogen peroxide; Oxidative stress.

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Conflict of interest statement

Competing Financial Interests: The authors have no competing financial interests

Figures

Figure 1
Figure 1
Global histone modification level in cells exposed to H2O2 for 3 hours with or without ascorbate pretreatment. (A)The ascorbate pretreatment was conducted at 250μM, 3 hours before sham or H2O2 was introduced to the BEAS-2B cell culture at 250μM for 3 hours. Histone H3 serves as a loading control. Band intensity relative to control band in each panel has been quantified and relative intensities are shown by the numbers below each panel. (B) Microscopy images of cells stained by carboxy-H2DCFDA. The cells were treated with 250 μM H2O2 for 3 hours, stained with carboxy-H2DCFDA (green) and Hoechst (blue); and observed using a confocal microscope. (C) GSH/GSSG ratio in cells exposed to H2O2 for 3 hours with or without ascorbate pretreatment.
Figure 2
Figure 2
Oxidative stress modulates the activity of histone modifying enzymes. (A) H2O2 inhibited the activity of histone H3K4 demethylase in vitro. The histone H3K4 demethylation assay was performed as described in Materials and Methods. The reaction mixture was incubated at 37°C for 1 hour. The intensity of the bands was quantified, and relative values relative to control condition were shown. + denotes 250uM for H2O2, 100μM for Fe(II), 1mM for ascorbate and 2mM for αKG; ++ denotes 200μM for Fe(II), 2mM for ascorbate and 4mM for αKG. *p<0.05 The lower panel shows stained protein gel image as a loading control.(B),(C) HDAC activity in cells exposed to H2O2 for 3 hours with or without ascorbate pretreatment.
Figure 3
Figure 3
Global histone modification level in long term oxidative stress and sustainability. (A) Global histone modification level in cells exposed to H2O2 for 3 weeks. (B) GSH/GSSG ratio in cells exposed to H2O2 for 3 weeks, *p<0.05. (C) Microscopy images of cells stained by carboxy-H2DCFDA. The cells were treated with 25 μM H2O2 for 3 weeks, stained with carboxy-H2DCFDA (green) and Hoechst (blue); and observed using a confocal microscope.
Figure 4
Figure 4
TET activity and DNA methylation under oxidative stress. (A)Global 5-mC level of BEAS-2B cells exposed to 150 μM H2O2 for 3 days. (B) Microscopy images of BEAS-2B cells stained by carboxy-H2DCFDA. The cells were treated with 150 μM H2O2 for 3 days, stained with carboxy-H2DCFDA (green) and Hoechst (blue). (C) GSH/GSSG ratio in BEAS-2B cells exposed to H2O2 for 3 days. (D). Fold enrichment of different areas of gpt gene after 5-hmC dependent DNA IP in G12 cells exposed to 150 μM H2O2 for 3 days when compared to control G12 cells. (E)TET activity in G12 cells. *p<0.05
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References

    1. Chia N, Wang L, Lu X, Senut MC, Brenner C, Ruden DM. Hypothesis: Environmental regulation of 5-hydroxymethyl-cytosine by oxidative stress. Epigenetics. 2011;6:7. - PubMed
    1. Costa M. Dioxygenases as targets of metals, hypoxia and oxidative stress during carcinogenesis. Molecular and genetic medicine. 2013;7:3.
    1. Berger SL. The complex language of chromatin regulation during transcription. Nature. 2007;447:407–412. - PubMed
    1. Whetstine JR, Nottke A, Lan F, Huarte M, Smolikov S, Chen Z, Spooner E, Li E, Zhang G, Colaiacovo M, Shi Y. Reversal of histone lysine trimethylation by the JMJD2 family of histone demethylases. Cell. 2006;125:467–481. - PubMed
    1. Wang J, Hevi S, Kurash JK, Lei H, Gay F, Bajko J, Su H, Sun W, Chang H, Xu G, Gaudet F, Li E, Chen T. The lysine demethylase LSD1 (KDM1) is required for maintenance of global DNA methylation. Nature genetics. 2009;41:125–129. - PubMed

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