title: Human mitochondrial pyrophosphatase: cDNA cloning and analysis of the gene in patients with mtDNA depletion syndromes.
abstract: Pyrophosphatases (PPases) catalyze the hydrolysis of inorganic pyrophosphate generated in several cellular enzymatic reactions. A novel human pyrophosphatase cDNA encoding a 334-amino-acid protein approximately 60% identical to the previously identified human cytosolic PPase was cloned and characterized. The novel enzyme, named PPase-2, was enzymatically active and catalyzed hydrolysis of pyrophosphate at a rate similar to that of the previously identified PPase-1. A functional mitochondrial import signal sequence was identified in the N-terminus of PPase-2, which targeted the enzyme to the mitochondrial matrix. The human pyrophosphatase 2 gene (PPase-2) was mapped to chromosome 4q25 and the 1.4-kb mRNA was ubiquitously expressed in human tissues, with highest levels in muscle, liver, and kidney. The yeast homologue of the mitochondrial PPase-2 is required for mitochondrial DNA maintenance and yeast cells lacking the enzyme exhibit mitochondrial DNA depletion. We sequenced the PPA2 gene in 13 patients with mitochondrial DNA depletion syndromes (MDS) of unknown cause to determine if mutations in the PPA2 gene of these patients were associated with this disease. No pathogenic mutations were identified in the PPA2 gene of these patients and we found no evidence that PPA2 gene mutations are a common cause of MDS in humans.
authors.affiliation: Division of Metabolic Disorders, Karolinska Institute, Stockholm, Sweden.
authors.forename: Sophie
authors.initials: S
authors.lastname: Curbo
authors.forename: Clotilde
authors.initials: C
authors.lastname: Lagier-Tourenne
authors.forename: Rosalba
authors.initials: R
authors.lastname: Carrozzo
authors.forename: Lluis
authors.initials: L
authors.lastname: Palenzuela
authors.forename: Simona
authors.initials: S
authors.lastname: Lucioli
authors.forename: Michio
authors.initials: M
authors.lastname: Hirano
authors.forename: Filippo
authors.initials: F
authors.lastname: Santorelli
authors.forename: Joaquin
authors.initials: J
authors.lastname: Arenas
authors.forename: Anna
authors.initials: A
authors.lastname: Karlsson
authors.forename: Magnus
authors.initials: M
authors.lastname: Johansson
mesh_terms: D000595:Amino Acid Sequence; D015152:Blotting, Northern; D002122:Calcium Chloride / Q000494:pharmacology; D045744:Cell Line, Tumor; D002874:Chromosome Mapping; D002894:Chromosomes, Human, Pair 4 / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D011756:Diphosphates / Q000378:metabolism; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D006868:Hydrolysis / Q000187:drug effects; D043564:Inorganic Pyrophosphatase / Q000235:genetics / Q000378:metabolism; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D017240:Mitochondrial Myopathies / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D024101:Mitochondrial Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D021382:Protein Sorting Signals / Q000235:genetics; D011755:Pyrophosphatases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013577:Syndrome; D014162:Transfection
chemical_list: D018076:DNA, Complementary; D011756:Diphosphates; D007527:Isoenzymes; D024101:Mitochondrial Proteins; D021382:Protein Sorting Signals; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D011755:Pyrophosphatases; D043564:Inorganic Pyrophosphatase; C509848:PPA2 protein, human; D002122:Calcium Chloride