Regulation of mitochondrial pyruvate uptake by alternative pyruvate carrier complexes

Bender, Tom; Pena, Gabrielle; Martinou, Jean‐Claude

doi:10.15252/embj.201490197

Cited by 120 publications

(142 citation statements)

References 39 publications

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“…MPC1L Can Replace MPC1 to Form a Functional MPC-To determine whether MPC1L is a functional MPC subunit, we used a complementation assay in a yeast strain lacking all MPC subunits (mpc1⌬/mpc2⌬/mpc3⌬). This strain fails to grow on synthetic medium lacking branched chain amino acids, for which mitochondrial pyruvate is an essential precursor in yeast (7,8,10). It was shown previously that this growth defect can be rescued by co-expression of the mouse MPC1 and MPC2 orthologs (7), and we have confirmed these results using human MPC1 and MPC2 (Fig.…”

Section: A a A T Ag L Tq R -------------------P Yg C --------A T T Tsupporting

confidence: 85%

“…These results indicate that MPC1L is an integral IMM protein, as was previously shown for MPC1 (7). In Saccharomyces cerevisiae, MPC1 is inserted in the IMM via two transmembrane segments, whereas the functionally redundant MPC2 and MPC3 subunits possess three transmembrane segments (10). We reasoned that a topological similarity between MPC1 and MPC1L in placental mammals would suggest a functional similarity between these two proteins.…”

Section: Mpc1l (Mpc1-like) Is a Mpc1 Paralog Specific To Placentalsupporting

confidence: 69%

“…MPC1-and MPC1L-containing MPCs Display Similar Pyruvate Import Efficiency-We next asked whether the physiological relevance for the existence of alternative MPC complexes in placental mammals lies in possible differences in pyruvate import efficiency, similar to that reported in yeast (10). The respirometry experiments shown in Fig.…”

Section: A a A T Ag L Tq R -------------------P Yg C --------A T T Tmentioning

confidence: 72%

“…The MPC was found to consist of two highly conserved protein subunits encoded in higher eukaryotes by the genes MPC1 and MPC2 (7,8). MPC1 and MPC2 are small transmembrane proteins of 109 and 127 amino acids, respectively, in humans, which physically interact with each other and form higher molecular weight complexes (8,10), the full composition of which remains to be fully elucidated.…”

mentioning

confidence: 99%

“…Study of the yeast MPC has revealed a switch in subunit composition depending on growth conditions. In fermentative conditions, the complex is composed of MPC1 and MPC2 (MPC FERM ), but in oxidative conditions MPC2 is replaced by the yeast specific MPC3 (MPC OX ), a MPC2 homolog that confers a greater capacity for pyruvate transport (10). In higher eukaryotes, no additional MPC subunits have been described to date.…”

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confidence: 99%

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MPC1-like Is a Placental Mammal-specific Mitochondrial Pyruvate Carrier Subunit Expressed in Postmeiotic Male Germ Cells

Vanderperre¹,

Cermakova²,

Escoffier

et al. 2016

Journal of Biological Chemistry

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Selective transport of pyruvate across the inner mitochondrial membrane by the mitochondrial pyruvate carrier (MPC) is a fundamental step that couples cytosolic and mitochondrial metabolism. The recent molecular identification of the MPC complex has revealed two interacting subunits, MPC1 and MPC2. Although in yeast, an additional subunit, MPC3, can functionally replace MPC2, no alternative MPC subunits have been described in higher eukaryotes. Here, we report for the first time the existence of a novel MPC subunit termed MPC1-like (MPC1L), which is present uniquely in placental mammals. MPC1L shares high sequence, structural, and topological homology with MPC1. In addition, we provide several lines of evidence to show that MPC1L is functionally equivalent to MPC1: 1) when co-expressed with MPC2, it rescues pyruvate import in a MPC-deleted yeast strain; 2) in mammalian cells, it can associate with MPC2 to form a functional carrier as assessed by bioluminescence resonance energy transfer; 3) in MPC1 depleted mouse embryonic fibroblasts, MPC1L rescues the loss of pyruvate-driven respiration and stabilizes MPC2 expression; and 4) MPC1-and MPC1L-mediated pyruvate imports show similar efficiency. However, we show that MPC1L has a highly specific expression pattern and is localized almost exclusively in testis and more specifically in postmeiotic spermatids and sperm cells. This is in marked contrast to MPC1/MPC2, which are ubiquitously expressed throughout the organism. To date, the biological importance of this alternative MPC complex during spermatogenesis in placental mammals remains unknown. Nevertheless, these findings open up new avenues for investigating the structure-function relationship within the MPC complex.The metabolic balance between glycolysis and oxidative phosphorylation (OXPHOS) 2 is of crucial importance in determining cell function and fate. Rapidly proliferating cells rely preferentially on glycolysis despite its lower yield of ATP, because glycolytic intermediates efficiently meet the anabolic needs for sustained cell growth. In rapidly dividing cells, this occurs even in the presence of oxygen, a process called aerobic glycolysis, or the Warburg effect (1, 2). Differentiated cells rely more heavily on OXPHOS to meet the energy demands associated with their specialization. In addition, a shift from glycolytic to oxidative metabolism is necessary for the differentiation process to occur (2-4), and conversely, a hallmark of cancer lies in the rewiring of cellular metabolism toward increased glycolytic flux (5). Pyruvate, the end product of glycolysis, is imported into the mitochondrial matrix where it fuels the TCA cycle and thereby provides electrons and reducing equivalents to the respiratory chain. Pyruvate-derived acetyl-CoA can also be used as an anabolic substrate for the synthesis of lipids and amino acids. Thus, elucidating the molecular mechanisms that determine the intracellular fate of pyruvate is important for understanding the control of cell metabolism and ultimately of cell funct...

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Section: A a A T Ag L Tq R -------------------P Yg C --------A T T Tsupporting

confidence: 85%

Section: Mpc1l (Mpc1-like) Is a Mpc1 Paralog Specific To Placentalsupporting

confidence: 69%

Section: A a A T Ag L Tq R -------------------P Yg C --------A T T Tmentioning

confidence: 72%

mentioning

confidence: 99%

mentioning

confidence: 99%

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MPC1-like Is a Placental Mammal-specific Mitochondrial Pyruvate Carrier Subunit Expressed in Postmeiotic Male Germ Cells

Vanderperre¹,

Cermakova²,

Escoffier

et al. 2016

Journal of Biological Chemistry

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Overexpression of the mitochondrial pyruvate carrier reduces lactate production and increases recombinant protein productivity in CHO cells

Bulté

Palomares

Parra

et al. 2020

Biotech & Bioengineering

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Chinese hamster ovary (CHO) cells are characterized by a low glucose catabolic efficiency, resulting in undesirable lactate production. Here, it is hypothesized that such low efficiency is determined by the transport of pyruvate into the mitochondria. The mitochondrial pyruvate carrier (MPC), responsible for introducing pyruvate into the mitochondria, is formed by two subunits, MPC1 and MPC2. Stable CHO cell lines, overexpressing the genes of both subunits, were constructed to facilitate the entry of pyruvate into the mitochondria and its incorporation into oxidative pathways. Significant overexpression of both genes, compared to the basal level of the control cells, was verified, and subcellular localization of both subunits in the mitochondria was confirmed. Kinetic evaluation of the best MPC overexpressing CHO cells showed a reduction of up to 50% in the overall yield of lactate production with respect to the control. An increase in specific growth rate and maximum viable cell concentration, as well as an increase of up to 40% on the maximum concentration of two recombinant model proteins transiently expressed (alkaline phosphatase or a monoclonal antibody), was also observed. Hybrid cybernetic modeling, that considered 89 reactions, 25 extracellular metabolites, and a network of 62 intracellular metabolites, explained that the best MPC overexpression case resulted in an increased metabolic flux across the mitochondrial membrane, activated a more balanced growth, and reduced the Warburg effect without compromising glucose consumption rate and maximum cell concentration. Overall, this study showed that transport of pyruvate into the mitochondria limits the efficiency of glucose oxidation, which can be overcome by a cell engineering approach.

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Mitochondrial Pyruvate Carriers are not Required for Adipogenesis but are Regulated by High‐Fat Feeding in Brown Adipose Tissue

Burrell

Richard

King

et al. 2020

Obesity

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Original Article OBESITY BIOLOGY AND INTEGRATED PHYSIOLOGY Study ImportanceWhat is already known?► Mitochondrial pyruvate carriers (MPCs) are essential for pyruvate to be transported into the mitochondria and utilized as substrate by the pyruvate dehydrogenase complex for energy production. What does this study add?► This study is novel because it demonstrates that MPCs are not required for the adipogenesis of 3T3-L1 cells, suggesting that other substrates may compensate for energy production during in vitro adipogenesis. ► This study is also novel because it shows that MPCs are regulated by diet-induced obesity in brown adipose tissue.Objective: The objectives of this study were to assess the role of mitochondrial pyruvate carriers (MPCs) in adipocyte development in vitro and determine whether MPCs are regulated in vivo by high-fat feeding in male and female C57BL/6J mice.Methods: This study utilized small interfering RNA-mediated knockdown to assess the requirement of MPC1 for adipogenesis in the 3T3-L1 model system. Treatment with UK-5099, a potent pharmacological MPC inhibitor, was also used to assess the loss of MPC activity. Western blot analysis was performed on adipose tissue samples from mice on a low-fat diet or a high-fat diet (HFD) for 12 weeks.Results: The loss of MPC expression via small interfering RNA-mediated knockdown or pharmacological inhibition did not affect adipogenesis of 3T3-L1 cells. In vivo studies indicated that expression of MPCs was significantly decreased in brown adipose tissue of male mice, but not female, on an HFD. Conclusions: Although MPCs are essential for pyruvate transport, MPCs are not required for adipogenesis in vitro, suggesting that other substrates can be used for energy production when the MPC complex is not functional. Also, a significant decrease in MPC1 and 2 expression occurred in brown fat, but not white fat, of male mice fed an HFD.Obesity (2020) 28, 293-302.

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Regulation of mitochondrial pyruvate uptake by alternative pyruvate carrier complexes

Cited by 120 publications

References 39 publications

MPC1-like Is a Placental Mammal-specific Mitochondrial Pyruvate Carrier Subunit Expressed in Postmeiotic Male Germ Cells

MPC1-like Is a Placental Mammal-specific Mitochondrial Pyruvate Carrier Subunit Expressed in Postmeiotic Male Germ Cells

Overexpression of the mitochondrial pyruvate carrier reduces lactate production and increases recombinant protein productivity in CHO cells

Mitochondrial Pyruvate Carriers are not Required for Adipogenesis but are Regulated by High‐Fat Feeding in Brown Adipose Tissue

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