Ca2+ metabolism in yeast cells and mitochondria

Carafoli, Ernesto; Balcavage, Walter X.; Lehninger, Albert L.; Mattoon, James R.

doi:10.1016/0005-2728(70)90057-5

Cited by 108 publications

(45 citation statements)

References 14 publications

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“…Neither does the Ca2+ uptake activity originate from the mitochondria, since it was measured in the presence of CCCP, an uncoupler of oxidative phosphorylation and a strong inhibitor of mitochondrial Ca2+ uptake [27]. In addition, it has been found that in yeast, Ca2+ is not taken up into the mitochondria [26]. Our results, therefore, strongly suggest that the observed pump is located in the endoplasmic reticulum, or other related intracellular compartments.…”

Section: Discussionmentioning

confidence: 55%

An intracellular ATP‐dependent calcium pump within the yeast Schizosaccharomyces pombe, encoded by the gene cta3

Halachmi

Ghislain

Eilam

1992

European Journal of Biochemistry

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We have permeabilized the plasma membranes of Schizosaccharomyces pombe cell with nystatin and measured ATP-dependent Ca2+ uptake in the presence of K N 0 3 and a protonophore in order to inhibit Ca2 + uptake into the vacuole. ATP-dependent Ca2 + accumulation into non-vacuolar Ca2 . However, the K , value for Ca2+ uptake by the vacuolar exchanger, 0.1 mM, seems to be too high to control Ca2+ concentration at submicromolar levels.Recently two genes for P-types ATPase, PMRl and PMR2, were identified in S. cerevisiae [15]. The deduced amino acid sequence of PMRl and PMR2 proteins was similar to that of animal Ca2+-ATPases. It was suggested that PMRl encodes a Ca2 + -ATPase that controls the secretory pathways. Our group has recently identified a novel P-type ATPase gene, eta3 in S. pombe [l 11. The deduced amino acid sequence shows that cta3 gene encodes a P-type ATPase. A null mutation in eta3 leads to higher levels of free cytosolic Ca2+, and lower amounts of sequestered and bound Ca2' . When the permeability of the plasma membrane was loosened by DEAE-dextran treatment, uptake of Ca2 + in the null mutant was lower than in the wild type. These physiological results supported the conclusion that eta3 protein is involved in Ca2 + transport and homeostasis and suggested localization of eta3 protein in intracellular membranes.In the present work, we investigated Ca2+ accumulation into intracellular compartments in nystatin-permeabilized cells, which are freely permeable to ATP. In the presence of inhibitors of Ca2+ uptake into the vacuole, an ATP-dependent Ca2+ uptake, which could be inhibited by vanadate was detected. These results are the first biochemical evidence of Ca2+-ATPase in yeast. ATP-dependent Ca2+-uptake is reduced in the yeast strain MG4 harboring a null allele of cta3.

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Section: Discussionmentioning

confidence: 55%

An intracellular ATP‐dependent calcium pump within the yeast Schizosaccharomyces pombe, encoded by the gene cta3

Halachmi

Ghislain

Eilam

1992

European Journal of Biochemistry

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“…Unlike mammalian mitochondria, yeast mitochondria apparently lack a high affinity Ca 2ϩ uniporter (67,68). Na ϩ /Ca 2ϩ antiporter activity also remains to be demonstrated in yeast mitochondria.…”

Section: Mitochondrial Namentioning

confidence: 98%

Identification of a Mitochondrial Na+/H+Exchanger

Numata

Petrecca

Lake

et al. 1998

Journal of Biological Chemistry

247

167

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The electroneutral exchange of protons for Na ؉ and K ؉ across the mitochondrial inner membrane contributes to organellar volume and Ca 2؉ homeostasis. The molecular nature of these transporters remains unknown. In this report, we characterize a novel gene (YDR456w; renamed NHA2) in Saccharomyces cerevisiae whose deduced protein sequence is homologous to members of the mammalian Na ؉ /H ؉ exchanger gene family. Fluorescence microscopy showed that a Nha2-green fluorescent protein chimera colocalizes with 4,6-diamidino-2-phenylindole staining of mitochondrial DNA. To assess the function of Nha2, we deleted the NHA2 gene by homologous disruption and found that benzamil-inhibitable, acid-activated 22 Na ؉ uptake into mitochondria was abolished in the mutant strain. It also showed retarded growth on nonfermentable carbon sources and severely reduced survival during the stationary phase of the cell cycle compared with the parental strain, consistent with a defect in aerobic metabolism. Sequence comparisons revealed that Nha2 has highest identity to a putative Na ؉ /H ؉ exchanger homologue (KIAA0267; renamed NHE6) in humans. Northern blot analysis demonstrated that NHE6 is ubiquitously expressed but is most abundant in mitochondrion-rich tissues such as brain, skeletal muscle, and heart. Fluorescence microscopy showed that a NHE6-green fluorescent protein chimera also accumulates in mitochondria of transfected HeLa cells. These data indicate that NHA2 and NHE6 encode homologous Na ؉ /H ؉ exchangers and suggest they may be important for mitochondrial function in lower and higher eukaryotes, respectively.

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“…Indeed, (i) the available inhibitors of channel function (e.g. RuR) lacked the protein specificity necessary for direct protein identification; (ii) Saccharomyces cerevisiae appears to have no RuR-sensitive mitochondrial Ca 2ϩ influx (83), thus preventing the use of yeast genetics; and (iii) in silico homology searches with known Ca 2ϩ channels yielded no successful hit. An alternative strategy was made possible by the seminal work of Mootha and co-workers (84), who constructed a mitochondrial gene data set (MitoCarta) by compiling an inventory of gene products with proven mitochondrial localization.…”

Section: The Discovery Of the Mcu: Finding The Needle In The Mitochonmentioning

confidence: 99%

The Mitochondrial Calcium Uniporter (MCU): Molecular Identity and Physiological Roles

Patrón

Raffaello

Granatiero

et al. 2013

Journal of Biological Chemistry

152

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The direct measurement of mitochondrial [Ca 2؉ ] with highly specific probes demonstrated that major swings in organellar [Ca 2؉ ] parallel the changes occurring in the cytosol and regulate processes as diverse as aerobic metabolism and cell death by necrosis and apoptosis. Despite great biological relevance, insight was limited by the complete lack of molecular understanding. The situation has changed, and new perspectives have emerged following the very recent identification of the mitochondrial Ca 2؉ uniporter, the channel allowing rapid Ca 2؉ accumulation across the inner mitochondrial membrane.

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Ca2+ metabolism in yeast cells and mitochondria

Cited by 108 publications

References 14 publications

An intracellular ATP‐dependent calcium pump within the yeast Schizosaccharomyces pombe, encoded by the gene cta3

An intracellular ATP‐dependent calcium pump within the yeast Schizosaccharomyces pombe, encoded by the gene cta3

Identification of a Mitochondrial Na+/H+Exchanger

The Mitochondrial Calcium Uniporter (MCU): Molecular Identity and Physiological Roles

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