Mutation in AAC2, equivalent to human adPEO-associated ANT1 mutations, lead to defective oxidative phosphorylation in Saccharomyces cerevisiae and affect mitochondrial DNA stability

Flavia Fontanesi, Luigi Palmieri, Pasquale Scarcia, Tiziana Lodi, Claudia Donnini, Anna Limongelli, Valeria Tiranti, Massimo Zeviani, Iliana Ferrero, Anna Maria Viola

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

Autosomal dominant and recessive forms of progressive external ophthalmoplegia (adPEO and arPEO) are mitochondrial disorders characterized by the presence of multiple deletions of mitochondrial DNA in affected tissues. Four adPEO-associated missense mutations have been identified in the ANT1 gene. In order to investigate their functional consequences on cellular physiology, we introduced three of them at equivalent positions in AAC2, the yeast orthologue of human ANT1. We demonstrate here that expression of the equivalent mutations in aac2-defective haploid strains of Saccharomyces cerevisiae results in (a) a marked growth defect on non-fermentable carbon sources, and (b) a concurrent reduction of the amount of mitochondrial cytochromes, cytochrome c oxidase activity and cellular respiration. The efficiency of ATP and ADP transport was variably affected by the different AAC2 mutations. However, irrespective of the absolute level of activity, the AAC2 pathogenic mutants showed a significant defect in ADP versus ATP transport compared with wild-type AAC2. In order to study whether a dominant phenotype, as in humans, could be observed, the aac2 mutant alleles were also inserted in combination with the endogenous wild-type AAC2 gene. The heteroallelic strains behaved as recessive for oxidative growth and petite-negative phenotype. In contrast, reduction in cytochrome content and increased mtDNA instability appeared to behave as dominant traits in heteroallelic strains. Our results indicate that S. cerevisiae is a suitable in vivo model to study the pathogenicity of the human ANT1 mutations and the pathophysiology leading to impairment of oxidative phosphorylation and damage of mtDNA integrity, as found in adPEO.

Original languageEnglish (US)
Pages (from-to)923-934
Number of pages12
JournalHuman Molecular Genetics
Volume13
Issue number9
DOIs
StatePublished - May 1 2004
Externally publishedYes

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Oxidative Phosphorylation
Mitochondrial DNA
Saccharomyces cerevisiae
Cytochromes
Adenosine Diphosphate
Mutation
Adenosine Triphosphate
Chronic Progressive External Ophthalmoplegia
Cell Respiration
Phenotype
Mitochondrial Diseases
Haploidy
Electron Transport Complex IV
Missense Mutation
Growth
Genes
Virulence
Carbon
Yeasts
Alleles

ASJC Scopus subject areas

  • Genetics

Cite this

Mutation in AAC2, equivalent to human adPEO-associated ANT1 mutations, lead to defective oxidative phosphorylation in Saccharomyces cerevisiae and affect mitochondrial DNA stability. / Fontanesi, Flavia; Palmieri, Luigi; Scarcia, Pasquale; Lodi, Tiziana; Donnini, Claudia; Limongelli, Anna; Tiranti, Valeria; Zeviani, Massimo; Ferrero, Iliana; Viola, Anna Maria.

In: Human Molecular Genetics, Vol. 13, No. 9, 01.05.2004, p. 923-934.

Research output: Contribution to journalArticle

Fontanesi, Flavia ; Palmieri, Luigi ; Scarcia, Pasquale ; Lodi, Tiziana ; Donnini, Claudia ; Limongelli, Anna ; Tiranti, Valeria ; Zeviani, Massimo ; Ferrero, Iliana ; Viola, Anna Maria. / Mutation in AAC2, equivalent to human adPEO-associated ANT1 mutations, lead to defective oxidative phosphorylation in Saccharomyces cerevisiae and affect mitochondrial DNA stability. In: Human Molecular Genetics. 2004 ; Vol. 13, No. 9. pp. 923-934.
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AU - Fontanesi, Flavia

AU - Palmieri, Luigi

AU - Scarcia, Pasquale

AU - Lodi, Tiziana

AU - Donnini, Claudia

AU - Limongelli, Anna

AU - Tiranti, Valeria

AU - Zeviani, Massimo

AU - Ferrero, Iliana

AU - Viola, Anna Maria

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AB - Autosomal dominant and recessive forms of progressive external ophthalmoplegia (adPEO and arPEO) are mitochondrial disorders characterized by the presence of multiple deletions of mitochondrial DNA in affected tissues. Four adPEO-associated missense mutations have been identified in the ANT1 gene. In order to investigate their functional consequences on cellular physiology, we introduced three of them at equivalent positions in AAC2, the yeast orthologue of human ANT1. We demonstrate here that expression of the equivalent mutations in aac2-defective haploid strains of Saccharomyces cerevisiae results in (a) a marked growth defect on non-fermentable carbon sources, and (b) a concurrent reduction of the amount of mitochondrial cytochromes, cytochrome c oxidase activity and cellular respiration. The efficiency of ATP and ADP transport was variably affected by the different AAC2 mutations. However, irrespective of the absolute level of activity, the AAC2 pathogenic mutants showed a significant defect in ADP versus ATP transport compared with wild-type AAC2. In order to study whether a dominant phenotype, as in humans, could be observed, the aac2 mutant alleles were also inserted in combination with the endogenous wild-type AAC2 gene. The heteroallelic strains behaved as recessive for oxidative growth and petite-negative phenotype. In contrast, reduction in cytochrome content and increased mtDNA instability appeared to behave as dominant traits in heteroallelic strains. Our results indicate that S. cerevisiae is a suitable in vivo model to study the pathogenicity of the human ANT1 mutations and the pathophysiology leading to impairment of oxidative phosphorylation and damage of mtDNA integrity, as found in adPEO.

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