SIRT3 and GCN5L regulation of NADP+- and NADPH-driven reactions of mitochondrial isocitrate dehydrogenase IDH2

Smolková, Katarína; Špačková, Jitka; Gotvaldová, Klára; Dvořák, Aleš; Křenková, Alena; Hubálek, Martin; Holendová, Blanka; Vı́tek, Libor; Ježek, Petr

doi:10.1038/s41598-020-65351-z

Cited by 14 publications

(12 citation statements)

References 43 publications

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“…The activity of PDH can be reduced in the absence of SIRT3 activity due to hyperacetylation of E1a subunit [ 22 ]. Similarly, the hyperacetylation of CS and ICDH2 leads to inhibition of enzyme activity [ 56 , 57 ]. Although aconitase is activated by acetylation [ 58 , 59 ], this enzyme is highly sensitive to free radical-induced damage [ 60 , 61 , 62 , 63 ].…”

Section: Discussionmentioning

confidence: 99%

Perturbed Brain Glucose Metabolism Caused by Absent SIRT3 Activity

Kristián

Karimi

Fearnow

et al. 2021

Cells

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Acetylation is a post-translational modification that regulates the activity of enzymes fundamentally involved in cellular and mitochondrial bioenergetic metabolism. NAD+ dependent deacetylase sirtuin 3 (SIRT3) is localized to mitochondria where it plays a key role in regulating acetylation of TCA cycle enzymes and the mitochondrial respiratory complexes. Although the SIRT3 target proteins in mitochondria have been identified, the effect of SIRT3 activity on mitochondrial glucose metabolism in the brain remains elusive. The impact of abolished SIRT3 activity on glucose metabolism was determined in SIRT3 knockout (KO) and wild type (WT) mice injected with [1,6-13C]glucose using ex vivo 13C-NMR spectroscopy. The 1H-NMR spectra and amino acid analysis showed no differences in the concentration of lactate, glutamate, alanine, succinate, or aspartate between SIRT3 KO and WT mice. However, glutamine, total creatine (Cr), and GABA were lower in SIRT3 KO brain. Incorporation of label from [1,6-13C]glucose metabolism into lactate or alanine was not affected in SIRT3 KO brain. However, the incorporation of the label into all isotopomers of glutamate, glutamine, GABA and aspartate was lower in SIRT3 KO brain, reflecting decreased activity of mitochondrial and TCA cycle metabolism in both neurons and astrocytes. This is most likely due to hyperacetylation of mitochondrial enzymes due to suppressed SIRT3 activity in the brain of SIRT3 KO mice. Thus, the absence of Sirt3 results in impaired mitochondrial oxidative energy metabolism and neurotransmitter synthesis in the brain. Since the SIRT3 activity is NAD+ dependent, these results might parallel changes in glucose metabolism under pathologic reduction in mitochondrial NAD+ pools.

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Section: Discussionmentioning

confidence: 99%

Perturbed Brain Glucose Metabolism Caused by Absent SIRT3 Activity

Kristián

Karimi

Fearnow

et al. 2021

Cells

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“…Additionally, in the oxidative mode (NADP + -dependent oxidative decarboxylation of IC into 2OG) IDH2 itself can be a donor of NADPH to the pool of enzymes performing the reduction of 2OG into 2HG, as demonstrated with purified IDH2 [9]. To provide evidence that IDH2 under certain conditions provides NADPH, we used IDH2 −/− MCF7 cells and overexpressed IDH2 WT or IDH2 R140Q (capable of neomorphic reduction of 2OG to 2HG, but not oxidative decarboxylation of IC or RC) to compare 2HG production at the background of IDH2 +/+ and −/− cells, respectively (Figure 4e, left).…”

Section: Role Of Nadph Balance In Idh2-produced 2hg Substrate and Cofactor Fluxmentioning

confidence: 99%

“…Subsequently, cells were harvested, washed with PBS, counted, and frozen until analyzed. The analysis was performed as described previously [9].…”

Section: Analytical Techniquesmentioning

confidence: 99%

“…Elevated concentrations of 2HG can be readily detected in the circulation of oncogenic patients [6,7]. Previously, we have demonstrated that 2HG can be formed by mitochondrial wild-type IDH2 despite the absence of any IDH2/1 mutations in breast cancer cells [8][9][10]. 2HG formation by wild-type IDH2 in breast cancer was also reported by Terunuma [11] and associated with the activity of the Myc oncogene, which augments glutaminolysis, and thus supports also reductive reactions catalyzed by IDH2, i.e., the reductive carboxylation (RC) into isocitrate (IC) and reduction of 2OG into 2HG.…”

Section: Introductionmentioning

confidence: 97%

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Biochemical Background in Mitochondria Affects 2HG Production by IDH2 and ADHFE1 in Breast Carcinoma

Špačková

Gotvaldová

Dvořák

et al. 2021

Cancers

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Mitochondrial production of 2-hydroxyglutarate (2HG) can be catalyzed by wild-type isocitrate dehydrogenase 2 (IDH2) and alcohol dehydrogenase, iron-containing 1 (ADHFE1). We investigated whether biochemical background and substrate concentration in breast cancer cells promote 2HG production. To estimate its role in 2HG production, we quantified 2HG levels and its enantiomers in breast cancer cells using analytical approaches for metabolomics. By manipulation of mitochondrial substrate fluxes using genetic and pharmacological approaches, we demonstrated the existence of active competition between 2HG producing enzymes, i.e., IDH2 and ADHFE1. Moreover, we showed that distinct fractions of IDH2 enzyme molecules operate in distinct oxido-reductive modes, providing NADPH and producing 2HG simultaneously. We have also detected 2HG release in the urine of breast cancer patients undergoing adjuvant therapy and detected a correlation with stages of breast carcinoma development. In summary, we provide a background for vital mitochondrial production of 2HG in breast cancer cells with outcomes towards cancer biology and possible future diagnosis of breast carcinoma.

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Section: Introductionmentioning

confidence: 97%

Biochemical Background in Mitochondria Affects 2HG Production by IDH2 and ADHFE1 in Breast Carcinoma

Špačková¹,

Gotvaldová²,

Dvořák³

et al. 2021

Preprint

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Mitochondrial production of 2-hydroxyglutarate (R-2HG) can be catalyzed by wild-type isocitrate dehydrogenase 2 (IDH2) and alcohol dehydrogenase, iron-containing 1 (ADHFE1). We investigated whether biochemical background and substrate concentration in breast cancer cells promote 2HG production. To estimate its role in 2HG production, we analyzed 2HG levels and its enantiomers in breast cancer cells using analytical approaches for metabolomics. By manipulation of mitochondrial substrate fluxes, including glutaminolysis, using genetic and pharmacological approaches we demonstrated the existence of an active competition between 2HG producing enzymes, i.e. IDH2 and ADHFE1. Moreover, we show that distinct fractions of IDH2 enzyme molecules operate in distinct oxido-reductive modes, providing NADPH and producing 2HG simultaneously. We have also detected 2HG release into breast cancer patient´s urine undergoing adjuvant therapy and detected a correlation with stages of breast carcinoma development. In summary, we provide a background for vital mitochondrial production of 2HG in breast cancer cells with outcomes towards cancer biology and possible future diagnosis of breast carcinoma.

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SIRT3 and GCN5L regulation of NADP+- and NADPH-driven reactions of mitochondrial isocitrate dehydrogenase IDH2

Cited by 14 publications

References 43 publications

Perturbed Brain Glucose Metabolism Caused by Absent SIRT3 Activity

Perturbed Brain Glucose Metabolism Caused by Absent SIRT3 Activity

Biochemical Background in Mitochondria Affects 2HG Production by IDH2 and ADHFE1 in Breast Carcinoma

Biochemical Background in Mitochondria Affects 2HG Production by IDH2 and ADHFE1 in Breast Carcinoma

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