{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2022,4,3]],"date-time":"2022-04-03T15:45:51Z","timestamp":1649000751356},"reference-count":58,"publisher":"Portland Press Ltd.","issue":"3","content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2012,12,15]]},"abstract":"<jats:p>Respiratory chain deficiency can result from alterations in mitochondrial and\/or cytosolic protein synthesis due to the dual genetic origin of mitochondrial oxidative phosphorylation. In the present paper we report a point mutation (D750G) in the bifunctional VARS (valyl-tRNA synthetase) of the fungus Neurospora crassa, associated with a temperature-sensitive phenotype. Analysis of the mutant strain revealed decreased steady-state levels of VARS and a clear reduction in the rate of mitochondrial protein synthesis. We observed a robust induction of the mitochondrial alternative oxidase with a concomitant decrease in the canonical respiratory pathway, namely in cytochrome b and aa3 content. Furthermore, the mutant strain accumulates the peripheral arm of complex I and depicts decreased levels of complexes III and IV, consistent with severe impairment of the mitochondrial respiratory chain. The phenotypic alterations of the mutant strain are observed at the permissive growth temperature and exacerbated upon increase of the temperature. Surprisingly, glucose-6-phosphate dehydrogenase activities were similar in the wild-type and mutant strains, whereas mitochondrial activities for succinate dehydrogenase and alternative NADH dehydrogenases were increased in the mutant strain, suggesting that the VARSD\u2212G mutation does not affect overall cytosolic protein synthesis. Expression of the wild-type vars gene rescues all of the mutant phenotypes, indicating that the VARSD\u2212G mutation is a loss-of-function mutation that results in a combined respiratory chain deficiency.<\/jats:p>","DOI":"10.1042\/bj20120963","type":"journal-article","created":{"date-parts":[[2012,9,10]],"date-time":"2012-09-10T08:42:16Z","timestamp":1347266536000},"page":"297-306","update-policy":"http:\/\/dx.doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Defective valyl-tRNA synthetase hampers the mitochondrial respiratory chain in <i>Neurospora crassa<\/i>"],"prefix":"10.1042","volume":"448","author":[{"given":"Margarida","family":"Duarte","sequence":"first","affiliation":[{"name":"IBMC (Instituto de Biologia Molecular e Celular), Universidade do Porto, Rua do Campo Alegre 823, 4150-180 Porto, Portugal"}]},{"given":"Arnaldo","family":"Videira","sequence":"additional","affiliation":[{"name":"IBMC (Instituto de Biologia Molecular e Celular), Universidade do Porto, Rua do Campo Alegre 823, 4150-180 Porto, Portugal"},{"name":"ICBAS (Instituto de Ci\u00eancias Biom\u00e9dicas de Abel Salazar), Universidade do Porto, Largo Prof. Abel Salazar 2, 4099-003 Porto, Portugal"}]}],"member":"288","published-online":{"date-parts":[[2012,11,21]]},"reference":[{"key":"2021112215274402500_B1","doi-asserted-by":"crossref","first-page":"1015","DOI":"10.1146\/annurev.bi.54.070185.005055","article-title":"The mitochondrial electron transport and oxidative phosphorylation system","volume":"54","author":"Hatefi","year":"1985","journal-title":"Annu. Rev. Biochem."},{"key":"2021112215274402500_B2","doi-asserted-by":"crossref","first-page":"737385","DOI":"10.1155\/2010\/737385","article-title":"Mitochondrial translation and beyond: processes implicated in combined oxidative phosphorylation deficiencies","volume":"2010","author":"Smits","year":"2010","journal-title":"J. Biomed. Biotechnol."},{"key":"2021112215274402500_B3","doi-asserted-by":"crossref","first-page":"1198","DOI":"10.1016\/j.bbabio.2011.06.010","article-title":"Human diseases with impaired mitochondrial protein synthesis","volume":"1807","author":"Rotig","year":"2011","journal-title":"Biochim. Biophys. Acta"},{"key":"2021112215274402500_B4","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1093\/brain\/awq320","article-title":"Nuclear factors involved in mitochondrial translation cause a subgroup of combined respiratory chain deficiency","volume":"134","author":"Kemp","year":"2011","journal-title":"Brain"},{"key":"2021112215274402500_B5","doi-asserted-by":"crossref","first-page":"715","DOI":"10.1146\/annurev.bi.62.070193.003435","article-title":"Cognition, mechanism, and evolutionary relationships in aminoacyl-tRNA synthetases","volume":"62","author":"Carter","year":"1993","journal-title":"Annu. Rev. Biochem."},{"key":"2021112215274402500_B6","doi-asserted-by":"crossref","first-page":"R536","DOI":"10.1016\/j.cub.2005.07.018","article-title":"Fidelity in protein synthesis","volume":"15","author":"Cochella","year":"2005","journal-title":"Curr. Biol."},{"key":"2021112215274402500_B7","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1016\/j.sbi.2011.11.008","article-title":"Structural analyses clarify the complex control of mistranslation by tRNA synthetases","volume":"22","author":"Guo","year":"2012","journal-title":"Curr. Opin. Struct. Biol."},{"key":"2021112215274402500_B8","doi-asserted-by":"crossref","first-page":"1293","DOI":"10.1086\/375039","article-title":"Glycyl tRNA synthetase mutations in Charcot-Marie-Tooth disease type\u00a02D and distal spinal muscular atrophy type\u00a0V","volume":"72","author":"Antonellis","year":"2003","journal-title":"Am. J. Hum. Genet."},{"key":"2021112215274402500_B9","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1146\/annurev.genom.9.081307.164204","article-title":"The role of aminoacyl-tRNA synthetases in genetic diseases","volume":"9","author":"Antonellis","year":"2008","journal-title":"Annu. Rev. Genomics Hum. Genet."},{"key":"2021112215274402500_B10","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1038\/ng1727","article-title":"Disrupted function and axonal distribution of mutant tyrosyl-tRNA synthetase in dominant intermediate Charcot-Marie-Tooth neuropathy","volume":"38","author":"Jordanova","year":"2006","journal-title":"Nat. Genet."},{"key":"2021112215274402500_B11","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1016\/j.ajhg.2009.12.005","article-title":"A major determinant for binding and aminoacylation of tRNA(Ala) in cytoplasmic alanyl-tRNA synthetase is mutated in dominant axonal Charcot-Marie-Tooth disease","volume":"86","author":"Latour","year":"2010","journal-title":"Am. J. Hum. Genet."},{"key":"2021112215274402500_B12","doi-asserted-by":"crossref","first-page":"560","DOI":"10.1016\/j.ajhg.2010.09.008","article-title":"Compound heterozygosity for loss-of-function lysyl-tRNA synthetase mutations in a patient with peripheral neuropathy","volume":"87","author":"McLaughlin","year":"2010","journal-title":"Am. J. Hum. Genet."},{"key":"2021112215274402500_B13","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1016\/j.ajhg.2010.12.010","article-title":"Mutations in the mitochondrial seryl-tRNA synthetase cause hyperuricemia, pulmonary hypertension, renal failure in infancy and alkalosis, HUPRA syndrome","volume":"88","author":"Belostotsky","year":"2011","journal-title":"Am. J. Hum. Genet."},{"key":"2021112215274402500_B14","doi-asserted-by":"crossref","first-page":"857","DOI":"10.1086\/521227","article-title":"Deleterious mutation in the mitochondrial arginyl-transfer RNA synthetase gene is associated with pontocerebellar hypoplasia","volume":"81","author":"Edvardson","year":"2007","journal-title":"Am. J. Hum. Genet."},{"key":"2021112215274402500_B15","doi-asserted-by":"crossref","first-page":"6543","DOI":"10.1073\/pnas.1103471108","article-title":"Mutations in mitochondrial histidyl tRNA synthetase HARS2 cause ovarian dysgenesis and sensorineural hearing loss of Perrault syndrome","volume":"108","author":"Pierce","year":"2011","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2021112215274402500_B16","doi-asserted-by":"crossref","first-page":"534","DOI":"10.1038\/ng2013","article-title":"Mitochondrial aspartyl-tRNA synthetase deficiency causes leukoencephalopathy with brain stem and spinal cord involvement and lactate elevation","volume":"39","author":"Scheper","year":"2007","journal-title":"Nat. Genet."},{"key":"2021112215274402500_B17","first-page":"4022","article-title":"The Neurospora crassa cyt-20 gene encodes cytosolic and mitochondrial valyl-tRNA synthetases and may have a second function in addition to protein synthesis","volume":"11","author":"Kubelik","year":"1991","journal-title":"Mol. Cell. Biol."},{"key":"2021112215274402500_B18","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1007\/BF00351683","article-title":"An yeast nuclear mutation conferring temperature-sensitivity to the mitochondrial tryptophanyl-tRNA synthetase","volume":"21","author":"Entrup","year":"1992","journal-title":"Curr. Genet."},{"key":"2021112215274402500_B19","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1093\/genetics\/93.3.539","article-title":"Isolation and characterization of temperature-sensitive respiratory mutants of Neurospora crassa","volume":"93","author":"Pittenger","year":"1979","journal-title":"Genetics"},{"key":"2021112215274402500_B20","doi-asserted-by":"crossref","first-page":"e10703","DOI":"10.1371\/journal.pone.0010703","article-title":"Characterization of the temperature-sensitive mutations un-7 and png-1\u00a0in Neurospora crassa","volume":"5","author":"Dieterle","year":"2010","journal-title":"PLoS ONE"},{"key":"2021112215274402500_B21","doi-asserted-by":"crossref","first-page":"37","DOI":"10.4148\/1941-4765.1089","article-title":"Demonstration that the Neurospora crassa mutation un-4 is a single nucleotide change in the tim16 gene encoding a subunit of the mitochondrial inner membrane translocase","volume":"55","author":"Wiest","year":"2008","journal-title":"Fungal Genet. Rep."},{"key":"2021112215274402500_B22","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1016\/0092-8674(87)90488-0","article-title":"A protein required for splicing group I introns in Neurospora mitochondria is mitochondrial tyrosyl-tRNA synthetase or a derivative thereof","volume":"50","author":"Akins","year":"1987","journal-title":"Cell"},{"key":"2021112215274402500_B23","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1016\/S0005-2728(02)00276-1","article-title":"From NADH to ubiquinone in Neurospora mitochondria","volume":"1555","author":"Videira","year":"2002","journal-title":"Biochim. Biophys. Acta"},{"key":"2021112215274402500_B24","doi-asserted-by":"crossref","first-page":"176","DOI":"10.1016\/S1087-1845(03)00002-1","article-title":"Alternative oxidase expression in Neurospora crassa","volume":"39","author":"Tanton","year":"2003","journal-title":"Fungal Genet. Biol."},{"key":"2021112215274402500_B25","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/0076-6879(71)17168-6","article-title":"Genetic and microbiological research techniques for Neurospora crassa","volume":"17A","author":"Davis","year":"1970","journal-title":"Methods Enzymol."},{"key":"2021112215274402500_B26","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1016\/S0065-2164(03)01010-4","article-title":"The Fungal Genetics Stock Center: from molds to molecules","volume":"52","author":"McCluskey","year":"2003","journal-title":"Adv. Appl. Microbiol."},{"key":"2021112215274402500_B27","doi-asserted-by":"crossref","first-page":"573","DOI":"10.1002\/j.1537-2197.1947.tb13032.x","article-title":"Neurospora V. A synthetic medium favoring sexual reproduction","volume":"34","author":"Westergaard","year":"1947","journal-title":"Am. J. Bot."},{"key":"2021112215274402500_B28","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1007\/s002940050277","article-title":"The Neurospora crassa cya-5 nuclear gene encodes a protein with a region of homology to the Saccharomyces cerevisiae PET309 protein and is required in a post-transcriptional step for the expression of the mitochondrially encoded COXI protein","volume":"32","author":"Coffin","year":"1997","journal-title":"Curr. Genet."},{"key":"2021112215274402500_B29","doi-asserted-by":"crossref","first-page":"3947","DOI":"10.1074\/jbc.M008199200","article-title":"The external calcium-dependent NADPH dehydrogenase from Neurospora crassa mitochondria","volume":"276","author":"Melo","year":"2001","journal-title":"J. Biol. Chem."},{"key":"2021112215274402500_B30","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1042\/bj1880141","article-title":"Stimulation of the alternative oxidase of Neurospora crassa by nucleoside phosphates","volume":"188","author":"Vanderleyden","year":"1980","journal-title":"Biochem. J."},{"key":"2021112215274402500_B31","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1016\/S1046-2023(02)00038-5","article-title":"Blue Native electrophoresis to study mitochondrial and other protein complexes","volume":"26","author":"Nijtmans","year":"2002","journal-title":"Methods"},{"key":"2021112215274402500_B32","doi-asserted-by":"crossref","first-page":"26453","DOI":"10.1074\/jbc.M402756200","article-title":"Supramolecular organization of cytochrome c oxidase- and alternative oxidase-dependent respiratory chains in the filamentous fungus Podospora anserina","volume":"279","author":"Krause","year":"2004","journal-title":"J. Biol. Chem."},{"key":"2021112215274402500_B33","volume-title":"Molecular Cloning: a Laboratory Manual","author":"Sambrook","year":"2001","edition":"3rd edn"},{"key":"2021112215274402500_B34","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1016\/0003-2697(76)90527-3","article-title":"A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding","volume":"72","author":"Bradford","year":"1976","journal-title":"Anal. Biochem."},{"key":"2021112215274402500_B35","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1111\/j.1432-1033.1985.tb09042.x","article-title":"Identification of the polypeptide encoded by the URF-1 gene of Neurospora crassa mtDNA","volume":"150","author":"Zauner","year":"1985","journal-title":"Eur. J. Biochem."},{"key":"2021112215274402500_B36","doi-asserted-by":"crossref","first-page":"4350","DOI":"10.1073\/pnas.76.9.4350","article-title":"Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications","volume":"76","author":"Towbin","year":"1979","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2021112215274402500_B37","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1016\/0003-2697(84)90320-8","article-title":"A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots","volume":"136","author":"Blake","year":"1984","journal-title":"Anal. Biochem."},{"key":"2021112215274402500_B38","doi-asserted-by":"crossref","first-page":"6347","DOI":"10.1016\/S0021-9258(18)61795-8","article-title":"Purification and partial characterization of glucose 6-phosphate dehydrogenase from Neurosopora crassa","volume":"246","author":"Scott","year":"1971","journal-title":"J. Biol. Chem."},{"key":"2021112215274402500_B39","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1093\/genetics\/71.4.521","article-title":"Isolation and classification of extranuclear mutants of Neurospora crassa","volume":"71","author":"Bertrand","year":"1972","journal-title":"Genetics"},{"key":"2021112215274402500_B40","doi-asserted-by":"crossref","first-page":"2391","DOI":"10.1128\/EC.00149-07","article-title":"Supramolecular organization of the respiratory chain in Neurospora crassa mitochondria","volume":"6","author":"Marques","year":"2007","journal-title":"Eukaryotic Cell"},{"key":"2021112215274402500_B41","doi-asserted-by":"crossref","first-page":"246","DOI":"10.1111\/j.1365-2958.2009.06643.x","article-title":"Effects of mitochondrial complex III disruption in the respiratory chain of Neurospora crassa","volume":"72","author":"Duarte","year":"2009","journal-title":"Mol. Microbiol."},{"key":"2021112215274402500_B42","doi-asserted-by":"crossref","first-page":"1091","DOI":"10.1016\/j.chembiol.2006.08.011","article-title":"Global effects of mistranslation from an editing defect in mammalian cells","volume":"13","author":"Nangle","year":"2006","journal-title":"Chem. Biol."},{"key":"2021112215274402500_B43","doi-asserted-by":"crossref","first-page":"e1002399","DOI":"10.1371\/journal.pgen.1002399","article-title":"Charcot-Marie-Tooth-linked mutant GARS is toxic to peripheral neurons independent of wild-type GARS levels","volume":"7","author":"Motley","year":"2011","journal-title":"PLoS Genet."},{"key":"2021112215274402500_B44","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1038\/nrg1606","article-title":"Mitochondrial DNA mutations in human disease","volume":"6","author":"Taylor","year":"2005","journal-title":"Nat. Rev. Genet."},{"key":"2021112215274402500_B45","doi-asserted-by":"crossref","first-page":"e1001288","DOI":"10.1371\/journal.pbio.1001288","article-title":"Mutations in the mitochondrial methionyl-tRNA synthetase cause a neurodegenerative phenotype in flies and a recessive ataxia (ARSAL) in humans","volume":"10","author":"Bayat","year":"2012","journal-title":"PLoS Biol."},{"key":"2021112215274402500_B46","doi-asserted-by":"crossref","first-page":"635","DOI":"10.1016\/j.ajhg.2011.04.006","article-title":"Exome sequencing identifies mitochondrial alanyl-tRNA synthetase mutations in infantile mitochondrial cardiomyopathy","volume":"88","author":"Gotz","year":"2011","journal-title":"Am. J. Hum. Genet."},{"key":"2021112215274402500_B47","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1016\/j.ajhg.2010.06.001","article-title":"Mutation of the mitochondrial tyrosyl-tRNA synthetase gene, YARS2, causes myopathy, lactic acidosis, and sideroblastic anemia-MLASA syndrome","volume":"87","author":"Riley","year":"2010","journal-title":"Am. J. Hum. Genet."},{"key":"2021112215274402500_B48","doi-asserted-by":"crossref","first-page":"1201","DOI":"10.1002\/humu.22098","article-title":"A novel mutation in YARS2 causes myopathy with lactic acidosis and sideroblastic anemia","volume":"33","author":"Sasarman","year":"2012","journal-title":"Hum. Mutat."},{"key":"2021112215274402500_B49","doi-asserted-by":"crossref","first-page":"793","DOI":"10.1016\/S0092-8674(00)00182-3","article-title":"Structural basis for double-sieve discrimination of L-valine from L-isoleucine and L-threonine by the complex of tRNA(Val) and valyl-tRNA synthetase","volume":"103","author":"Fukai","year":"2000","journal-title":"Cell"},{"key":"2021112215274402500_B50","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1261\/rna.2760703","article-title":"Mechanism of molecular interactions for tRNA(Val) recognition by valyl-tRNA synthetase","volume":"9","author":"Fukai","year":"2003","journal-title":"RNA"},{"key":"2021112215274402500_B51","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1016\/S0167-4781(99)00240-7","article-title":"Expression and characterization of the human mitochondrial leucyl-tRNA synthetase","volume":"1490","author":"Bullard","year":"2000","journal-title":"Biochim. Biophys. Acta"},{"key":"2021112215274402500_B52","doi-asserted-by":"crossref","first-page":"16820","DOI":"10.1074\/jbc.275.22.16820","article-title":"Identification and characterization of human mitochondrial tryptophanyl-tRNA synthetase","volume":"275","author":"Jorgensen","year":"2000","journal-title":"J. Biol. Chem."},{"key":"2021112215274402500_B53","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1038\/nature05096","article-title":"Editing-defective tRNA synthetase causes protein misfolding and neurodegeneration","volume":"443","author":"Lee","year":"2006","journal-title":"Nature"},{"key":"2021112215274402500_B54","doi-asserted-by":"crossref","first-page":"244","DOI":"10.1002\/humu.21635","article-title":"A recurrent loss-of-function alanyl-tRNA synthetase (AARS) mutation in patients with Charcot-Marie-Tooth disease type\u00a02N (CMT2N)","volume":"33","author":"McLaughlin","year":"2012","journal-title":"Hum. Mutat."},{"key":"2021112215274402500_B55","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1016\/j.mito.2011.04.003","article-title":"Mutation in the mitochondrial tRNA(Val) causes mitochondrial encephalopathy, lactic acidosis and stroke-like episodes","volume":"11","author":"Glatz","year":"2011","journal-title":"Mitochondrion"},{"key":"2021112215274402500_B56","doi-asserted-by":"crossref","first-page":"3065","DOI":"10.1093\/nar\/gkn147","article-title":"Overexpression of human mitochondrial valyl tRNA synthetase can partially restore levels of cognate mt-tRNAVal carrying the pathogenic C25U mutation","volume":"36","author":"Rorbach","year":"2008","journal-title":"Nucleic Acids Res."},{"key":"2021112215274402500_B57","doi-asserted-by":"crossref","first-page":"1629","DOI":"10.1128\/jb.94.5.1629-1637.1967","article-title":"Multiple alterations in metabolite uptake in a mutant of Neurospora crassa","volume":"94","author":"Kappy","year":"1967","journal-title":"J. Bacteriol."},{"key":"2021112215274402500_B58","doi-asserted-by":"crossref","first-page":"11782","DOI":"10.1073\/pnas.0905339106","article-title":"Dominant mutations in the tyrosyl-tRNA synthetase gene recapitulate in Drosophila features of human Charcot-Marie-Tooth neuropathy","volume":"106","author":"Storkebaum","year":"2009","journal-title":"Proc. Natl. Acad. Sci. U.S.A."}],"container-title":["Biochemical Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/portlandpress.com\/biochemj\/article-pdf\/448\/3\/297\/672513\/bj4480297.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/portlandpress.com\/biochemj\/article-pdf\/448\/3\/297\/672513\/bj4480297.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,11,22]],"date-time":"2021-11-22T21:03:21Z","timestamp":1637615001000},"score":1,"resource":{"primary":{"URL":"https:\/\/portlandpress.com\/biochemj\/article\/448\/3\/297\/45934\/Defective-valyl-tRNA-synthetase-hampers-the"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2012,11,21]]},"references-count":58,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2012,12,15]]}},"URL":"https:\/\/doi.org\/10.1042\/bj20120963","relation":{},"ISSN":["0264-6021","1470-8728"],"issn-type":[{"value":"0264-6021","type":"print"},{"value":"1470-8728","type":"electronic"}],"subject":[],"published":{"date-parts":[[2012,11,21]]}}}