Protein Hydroxylation Catalyzed by 2-Oxoglutarate-dependent Oxygenases

Markolovic, Suzana; Wilkins, Sarah E.; Schofield, Christopher J.

doi:10.1074/jbc.r115.662627

Cited by 150 publications

(135 citation statements)

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“…These studies reveal that vitamin C plays a role in reducing inactive iron and maintaining continued enzyme cycling. Because Tet dioxygenases are also dependent on iron and 2-oxoglutarate (32,58), and the catalytic cycle of the iron-and 2-oxoglutarate-dependent dioxygenase is highly conserved (59), it is tempting to speculate that vitamin C might help Tet enzymes like collagen prolyl-4-hydroxylase enzymes. Indeed, although the hydroxylation of 5mC into 5hmC was initiated and 5hmC was enriched in the Foxp3 CNS2 regions of iTregs, further oxidation reaction was not completed (Fig.…”

Section: Discussionmentioning

confidence: 99%

Vitamin C Facilitates Demethylation of the Foxp3 Enhancer in a Tet-Dependent Manner

Nair

Song

2016

The Journal of Immunology

147

100

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Demethylation of CpG motifs in the Foxp3 intronic element, conserved noncoding sequence 2 (CNS2), is indispensable for the stable expression of Foxp3 in regulatory T cells (Tregs). In this study, we found that vitamin C induces CNS2 demethylation in Tregs in a ten-eleven-translocation 2 (Tet2)-dependent manner. The CpG motifs of CNS2 in Tregs generated in vitro by TGF-β (iTregs), which were methylated originally, became demethylated after vitamin C treatment. The conversion of 5-methylcytosin into 5-hydroxymethylcytosin was more efficient, and the methyl group from the CpG motifs of Foxp3 CNS2 was erased rapidly in iTregs treated with vitamin C. The effect of vitamin C disappeared in Tet2−/− iTregs. Furthermore, CNS2 in peripheral Tregs in vivo, which were demethylated originally, became methylated after treatment with a sodium-dependent vitamin C transporter inhibitor, sulfinpyrazone. Finally, CNS2 demethylation in thymic Tregs was also impaired in Tet2−/− mice, but not in wild type mice, when they were treated with sulfinpyrazone. Collectively, vitamin C was required for the CNS2 demethylation mediated by Tet proteins, which was essential for Foxp3 expression. Our findings indicate that environmental factors, such as nutrients, could bring about changes in immune homeostasis through epigenetic mechanisms.

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Section: Discussionmentioning

confidence: 99%

Vitamin C Facilitates Demethylation of the Foxp3 Enhancer in a Tet-Dependent Manner

Nair

Song

2016

The Journal of Immunology

147

100

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“…In addition to acetylation, protein lysine modifications by possibly non-enzymatic malonylation, succinylation and glutarylation utilize mitochondrial intermediates for regulation of protein functions, reversible by the de-acylase Sirt5 [3][4][5]. Alpha-ketoglutarate is a substrate for prolyl hydroxylases, including regulators of hypoxia-inducible factors (HIFs), and dioxygenases with DNA and histone demethylation functions [6,7]. Mitochondria are a major cellular pool of calcium ions, with physiologic functions in bioenergetics and retrograde signaling, and pathophysiologic roles in necrotic cell death [8][9][10].…”

Section: Overview Of Mitochondrial Functionsmentioning

confidence: 99%

Mitochondrial functions in stem cells

Margineantu

Hockenbery

2016

Current Opinion in Genetics & Development

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“…One important class of enzymes that utilizes αKG—and thus potentially susceptible to inhibition by 2HG—is the non-heme iron- and αKG-dependent dioxygenases, a family of enzymes with over 80 different members. 21–24 2HG has been demonstrated to inhibit several αKG-dependent enzymes, such as histone demethylases, prolyl hydroxylases, the TET family 5-methlycytosine (5mC) hydroxylases, 7 and some of the AlkB family DNA repair enzymes, such as ALKBH2 and ALKBH3. 8,25 However, in the case of the AlkB proteins, previous in vitro studies have only provided the IC 50 values of 2HG on the enzymes and the results have not been extrapolated to physiologically relevant concentrations of 2HG and αKG in cancer cells.…”

Section: Introductionmentioning

confidence: 99%

Oncometabolites d- and l-2-Hydroxyglutarate Inhibit the AlkB Family DNA Repair Enzymes under Physiological Conditions

Chen

Bian

Tang

et al. 2017

Chem. Res. Toxicol.

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Cancer-associated mutations often lead to perturbed cellular energy metabolism and accumulation of potentially harmful oncometabolites. One example is the chiral molecule 2-hydroxyglutarate (2HG); its two stereoisomers (D- and L-2HG) have been found with abnormally high concentrations in tumors featuring anomalous metabolic pathways. 2HG has been demonstrated to competitively inhibit several α-ketoglutarate (αKG)- and non-heme iron-dependent dioxygenases, including some of the AlkB family DNA repair enzymes, such as ALKBH2 and ALKBH3. However, previous studies have only provided the IC50 values of D-2HG on the enzymes and the results have not been correlated to physiologically relevant concentrations of 2HG and αKG in cancer cells. In this work, we carried out detailed kinetic analyses of DNA repair reactions catalyzed by ALKBH2, ALKBH3 and the bacterial AlkB in the presence of D- and L-2HG in both double and single stranded DNA contexts. We determined kinetic parameters of inhibition, including kcat, KM, and Ki. We also correlated the relative concentrations of 2HG and αKG previously measured in tumor cells with the inhibitory effect of 2HG on the AlkB family enzymes. Both D- and L-2HG significantly inhibited the human DNA repair enzymes ALKBH2 and ALKBH3 under pathologically relevant concentrations (73–88% for D-2HG and 31–58% for L-2HG inhibition). This work provides a new perspective that the elevation of either D- or L-2HG in cancer cells may contribute to an increased mutation rate by inhibiting the DNA repair carried out by the AlkB family enzymes and thus exacerbate the genesis and progression of tumors.

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Protein Hydroxylation Catalyzed by 2-Oxoglutarate-dependent Oxygenases

Cited by 150 publications

References 100 publications

Vitamin C Facilitates Demethylation of the Foxp3 Enhancer in a Tet-Dependent Manner

Vitamin C Facilitates Demethylation of the Foxp3 Enhancer in a Tet-Dependent Manner

Mitochondrial functions in stem cells

Oncometabolites d- and l-2-Hydroxyglutarate Inhibit the AlkB Family DNA Repair Enzymes under Physiological Conditions

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