title: The Saccharomyces cerevisiae gene YPR011c encodes a mitochondrial transporter of adenosine 5'-phosphosulfate and 3'-phospho-adenosine 5'-phosphosulfate.
abstract: The genome of Saccharomyces cerevisiae contains 35 members of the mitochondrial carrier family, nearly all of which have been functionally characterized. In this study, the identification of the mitochondrial carrier for adenosine 5'-phosphosulfate (APS) is described. The corresponding gene (YPR011c) was overexpressed in bacteria. The purified protein was reconstituted into phospholipid vesicles and its transport properties and kinetic parameters were characterized. It transported APS, 3'-phospho-adenosine 5'-phosphosulfate, sulfate and phosphate almost exclusively by a counter-exchange mechanism. Transport was saturable and inhibited by bongkrekic acid and other inhibitors. To investigate the physiological significance of this carrier in S. cerevisiae, mutants were subjected to thermal shock at 45°C in the presence of sulfate and in the absence of methionine. At 45°C cells lacking YPR011c, engineered cells (in which APS is produced only in mitochondria) and more so the latter cells, in which the exit of mitochondrial APS is prevented by the absence of YPR011cp, were less thermotolerant. Moreover, at the same temperature all these cells contained less methionine and total glutathione than wild-type cells. Our results show that S. cerevisiae mitochondria are equipped with a transporter for APS and that YPR011cp-mediated mitochondrial transport of APS occurs in S. cerevisiae under thermal stress conditions.
authors.affiliation: Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari, via Orabona 4, 70125 Bari, Italy; Center of Excellence in Comparative Genomics, University of Bari, Italy.
authors.forename: Simona
authors.initials: S
authors.lastname: Todisco
authors.affiliation: Department of Sciences, University of Basilicata, via N. Sauro 85, 85100 Potenza, Italy.
authors.forename: Maria Antonietta
authors.initials: MA
authors.lastname: Di Noia
authors.affiliation: Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari, via Orabona 4, 70125 Bari, Italy; Center of Excellence in Comparative Genomics, University of Bari, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Castegna
authors.affiliation: Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari, via Orabona 4, 70125 Bari, Italy; CNR Institute of Biomembranes and Bioenergetics, via Amendola 165/A, 70126 Bari, Italy.
authors.forename: Francesco Massimo
authors.initials: FM
authors.lastname: Lasorsa
authors.affiliation: Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari, via Orabona 4, 70125 Bari, Italy; CNR Institute of Biomembranes and Bioenergetics, via Amendola 165/A, 70126 Bari, Italy.
authors.forename: Eleonora
authors.initials: E
authors.lastname: Paradies
authors.affiliation: Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari, via Orabona 4, 70125 Bari, Italy; Center of Excellence in Comparative Genomics, University of Bari, Italy. Electronic address: ferdpalmieri@gmail.com.
authors.forename: Ferdinando
authors.initials: F
authors.lastname: Palmieri
mesh_terms: D000222:Adaptation, Physiological; D000250:Adenosine Phosphosulfate / Q000378:metabolism*; D001692:Biological Transport / Q000235:genetics; D003065:Coenzyme A / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D005800:Genes, Fungal / Q000235:genetics*; D005816:Genetic Complementation Test; D005978:Glutathione / Q000378:metabolism; D007700:Kinetics; D008715:Methionine / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism*; D050505:Mutant Proteins / Q000378:metabolism; D010724:Phosphoadenosine Phosphosulfate / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism*; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D050505:Mutant Proteins; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; C586987:YPR011c protein, s cerevisiae; D010724:Phosphoadenosine Phosphosulfate; D000250:Adenosine Phosphosulfate; D008715:Methionine; D005978:Glutathione; D003065:Coenzyme A
keywords: 3′-phospho-adenosine 5′-phosphosulfate; APS; Adenosine 5′-phosphosulfate; ApppA; CoA; MCF; Mitochondrial carrier; Mitochondrion; P(1),P(3)-di(adenosine-5′) triphosphate; PAP; PAPS; Saccharomyces cerevisiae; Thermotolerance; Transporter; YPR011cp; adenosine 3′,5′-diphosphate; adenosine 5′-phosphosulfate; coenzyme A; mitochondrial Met3p; mitochondrial carrier family; mtMet3p; the protein encoded by YPR011c