Analysis of Mitochondrial Proteome of Cybrid Cells Harbouring a Truncative Mitochondrial DNA Mutation in Respiratory Complex I.

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Abstract

Transmitochondrial cytoplasmic hybrids (cybrids) are well established model systems to reveal the effects ofmitochondrial DNA (mtDNA) mutations on cell metabolism excluding the interferences of a different nuclearbackground. The m.3571insC mutation in the MTND1 gene of respiratory complex I (CI) is commonlydetected in oncocytic tumors, in which it causes a severe CI dysfunction leading to an energetic impairmentwhen present above 83% mutant load. To assess whether the energetic deficit may alter the mitochondrialproteome, OS-78 and OS-93 cybrid cell lines bearing two different degrees of the m.3571insC mutation(78% and 92.8%, respectively) and control cybrids bearing wild-type mtDNA (CC) were analyzed. Twodimensionalelectrophoresis and mass spectrometry revealed significant alterations only in cybrids abovethe threshold (OS-93). All differentially expressed proteins are decreased. In particular, the levels of thepyruvate dehydrogenase E1 chain B subunit (E1b), of lipoamide dehydrogenase (E3), the enzymecomponent of pyruvate and 2-oxoglutarate dehydrogenase complexes, and of lactate dehydrogenase B(LDHB) were reduced. Moreover, a significant decrease of the pyruvate dehydrogenase complex activitywas found when OS-93 cybrid cells were grown in galactose medium, a metabolic condition that forcescells to use respiration. These results demonstrate that the energetic impairment caused by the almosthomoplasmic m.3571insC mutation perturbs cellular metabolism leading to a decreased steady statelevel of components of very important mitochondrial NAD-dependent dehydrogenases.

Autore Pugliese

Tutti gli autori

  • C. Musicco; A. Cormio; M.A. Calvaruso; L. Iommarini; G. Gasparre; A.M. Porcelli; A.M. Timperio; L. Zolla; M.N. Gadaleta

Titolo volume/Rivista

Molecular bioSystems

Anno di pubblicazione

2014

ISSN

1742-206X

ISBN

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