The cytochrome b p.278Y>C mutation causative of a multisystem disorder enhances superoxide production and alters supramolecular interactions of respiratory chain complexes

Anna Ghelli; Concetta V Tropeano; Maria Antonietta Calvaruso; Alessandra Marchesini; Luisa Iommarini; Anna Maria Porcelli; Claudia Zanna; Vera De Nardo; Andrea Martinuzzi; Flemming Wibrand; John Vissing; Ivana Kurelac; Giuseppe Gasparre; Nur Selamoglu; Fevzi Daldal; Michela Rugolo

doi:10.1093/hmg/ddt067

The cytochrome b p.278Y>C mutation causative of a multisystem disorder enhances superoxide production and alters supramolecular interactions of respiratory chain complexes

Anna Ghelli, Concetta V Tropeano, Maria Antonietta Calvaruso, Alessandra Marchesini, Luisa Iommarini, Anna Maria Porcelli, Claudia Zanna, Vera De Nardo, Andrea Martinuzzi, Flemming Wibrand, John Vissing, Ivana Kurelac, Giuseppe Gasparre, Nur Selamoglu, Fevzi Daldal, Michela Rugolo

Department of Clinical Medicine

34 Citations (Scopus)

Abstract

Cytochrome b is the only mtDNA-encoded subunit of the mitochondrial complex III (CIII), the functional bottleneck of the respiratory chain. Previously, the human cytochrome b missense mutation m.15579A>G, which substitutes the Tyr 278 with Cys (p.278Y>C), was identified in a patient with severe exercise intolerance and multisystem manifestations. In this study, we characterized the biochemical properties of cybrids carrying this mutation and report that the homoplasmic p.278Y>C mutation caused a dramatic reduction in the CIII activity and in CIII-driven mitochondrial ATP synthesis. However, the CI, CI 1 CIII and CII 1 CIII activities and the rate of ATP synthesis driven by the CI or CII substrate were only partially reduced or unaffected. Consistent with these findings, mutated cybrids maintained the mitochondrial membrane potential in the presence of oligomycin, indicating that it originated from the respiratory electron transport chain. The p.278Y>C mutation enhanced superoxide production, as indicated by direct measurements in mitochondria and by the imbalance of glutathione homeostasis in intact cybrids. Remarkably, although the assembly of CI or CIII was not affected, the examination of respiratory supercomplexes revealed that the amounts of CIII dimer and III2IV1 were reduced, whereas those of I1III2IVn slightly increased. We therefore suggest that the deleterious effects of p.278Y>C mutation on cytochrome b are palliated when CIII is assembled into the supercomplexes I1III2IVn, in contrast to when it is found alone. These findings underline the importance of supramolecular interactions between complexes for maintaining a basal respiratory chain activity and shed light to the molecular basis of disease manifestations associated with this mutation.

Original language	English
Journal	Human Molecular Genetics
Volume	22
Issue number	11
Pages (from-to)	2141-2151
Number of pages	11
ISSN	0964-6906
DOIs	https://doi.org/10.1093/hmg/ddt067
Publication status	Published - Jun 2013

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Ghelli, A., Tropeano, C. V., Calvaruso, M. A., Marchesini, A., Iommarini, L., Porcelli, A. M., Zanna, C., De Nardo, V., Martinuzzi, A., Wibrand, F., Vissing, J., Kurelac, I., Gasparre, G., Selamoglu, N., Daldal, F., & Rugolo, M. (2013). The cytochrome b p.278Y>C mutation causative of a multisystem disorder enhances superoxide production and alters supramolecular interactions of respiratory chain complexes. Human Molecular Genetics, 22(11), 2141-2151. https://doi.org/10.1093/hmg/ddt067

The cytochrome b p.278Y>C mutation causative of a multisystem disorder enhances superoxide production and alters supramolecular interactions of respiratory chain complexes. / Ghelli, Anna; Tropeano, Concetta V; Calvaruso, Maria Antonietta et al.

In: Human Molecular Genetics, Vol. 22, No. 11, 06.2013, p. 2141-2151.

Research output: Contribution to journal › Journal article › Research › peer-review

Ghelli, A, Tropeano, CV, Calvaruso, MA, Marchesini, A, Iommarini, L, Porcelli, AM, Zanna, C, De Nardo, V, Martinuzzi, A, Wibrand, F, Vissing, J, Kurelac, I, Gasparre, G, Selamoglu, N, Daldal, F & Rugolo, M 2013, 'The cytochrome b p.278Y>C mutation causative of a multisystem disorder enhances superoxide production and alters supramolecular interactions of respiratory chain complexes', Human Molecular Genetics, vol. 22, no. 11, pp. 2141-2151. https://doi.org/10.1093/hmg/ddt067

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title = "The cytochrome b p.278Y>C mutation causative of a multisystem disorder enhances superoxide production and alters supramolecular interactions of respiratory chain complexes",

abstract = "Cytochrome b is the only mtDNA-encoded subunit of the mitochondrial complex III (CIII), the functional bottleneck of the respiratory chain. Previously, the human cytochrome b missense mutation m.15579A>G, which substitutes the Tyr 278 with Cys (p.278Y>C), was identified in a patient with severe exercise intolerance and multisystem manifestations. In this study, we characterized the biochemical properties of cybrids carrying this mutation and report that the homoplasmic p.278Y>C mutation caused a dramatic reduction in the CIII activity and in CIII-driven mitochondrial ATP synthesis. However, the CI, CI 1 CIII and CII 1 CIII activities and the rate of ATP synthesis driven by the CI or CII substrate were only partially reduced or unaffected. Consistent with these findings, mutated cybrids maintained the mitochondrial membrane potential in the presence of oligomycin, indicating that it originated from the respiratory electron transport chain. The p.278Y>C mutation enhanced superoxide production, as indicated by direct measurements in mitochondria and by the imbalance of glutathione homeostasis in intact cybrids. Remarkably, although the assembly of CI or CIII was not affected, the examination of respiratory supercomplexes revealed that the amounts of CIII dimer and III2IV1 were reduced, whereas those of I1III2IVn slightly increased. We therefore suggest that the deleterious effects of p.278Y>C mutation on cytochrome b are palliated when CIII is assembled into the supercomplexes I1III2IVn, in contrast to when it is found alone. These findings underline the importance of supramolecular interactions between complexes for maintaining a basal respiratory chain activity and shed light to the molecular basis of disease manifestations associated with this mutation.",

author = "Anna Ghelli and Tropeano, {Concetta V} and Calvaruso, {Maria Antonietta} and Alessandra Marchesini and Luisa Iommarini and Porcelli, {Anna Maria} and Claudia Zanna and {De Nardo}, Vera and Andrea Martinuzzi and Flemming Wibrand and John Vissing and Ivana Kurelac and Giuseppe Gasparre and Nur Selamoglu and Fevzi Daldal and Michela Rugolo",

year = "2013",

month = jun,

doi = "10.1093/hmg/ddt067",

language = "English",

volume = "22",

pages = "2141--2151",

journal = "Human Molecular Genetics",

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T1 - The cytochrome b p.278Y>C mutation causative of a multisystem disorder enhances superoxide production and alters supramolecular interactions of respiratory chain complexes

AU - Ghelli, Anna

AU - Tropeano, Concetta V

AU - Calvaruso, Maria Antonietta

AU - Marchesini, Alessandra

AU - Iommarini, Luisa

AU - Porcelli, Anna Maria

AU - Zanna, Claudia

AU - De Nardo, Vera

AU - Martinuzzi, Andrea

AU - Wibrand, Flemming

AU - Vissing, John

AU - Kurelac, Ivana

AU - Gasparre, Giuseppe

AU - Selamoglu, Nur

AU - Daldal, Fevzi

AU - Rugolo, Michela

PY - 2013/6

Y1 - 2013/6

N2 - Cytochrome b is the only mtDNA-encoded subunit of the mitochondrial complex III (CIII), the functional bottleneck of the respiratory chain. Previously, the human cytochrome b missense mutation m.15579A>G, which substitutes the Tyr 278 with Cys (p.278Y>C), was identified in a patient with severe exercise intolerance and multisystem manifestations. In this study, we characterized the biochemical properties of cybrids carrying this mutation and report that the homoplasmic p.278Y>C mutation caused a dramatic reduction in the CIII activity and in CIII-driven mitochondrial ATP synthesis. However, the CI, CI 1 CIII and CII 1 CIII activities and the rate of ATP synthesis driven by the CI or CII substrate were only partially reduced or unaffected. Consistent with these findings, mutated cybrids maintained the mitochondrial membrane potential in the presence of oligomycin, indicating that it originated from the respiratory electron transport chain. The p.278Y>C mutation enhanced superoxide production, as indicated by direct measurements in mitochondria and by the imbalance of glutathione homeostasis in intact cybrids. Remarkably, although the assembly of CI or CIII was not affected, the examination of respiratory supercomplexes revealed that the amounts of CIII dimer and III2IV1 were reduced, whereas those of I1III2IVn slightly increased. We therefore suggest that the deleterious effects of p.278Y>C mutation on cytochrome b are palliated when CIII is assembled into the supercomplexes I1III2IVn, in contrast to when it is found alone. These findings underline the importance of supramolecular interactions between complexes for maintaining a basal respiratory chain activity and shed light to the molecular basis of disease manifestations associated with this mutation.

AB - Cytochrome b is the only mtDNA-encoded subunit of the mitochondrial complex III (CIII), the functional bottleneck of the respiratory chain. Previously, the human cytochrome b missense mutation m.15579A>G, which substitutes the Tyr 278 with Cys (p.278Y>C), was identified in a patient with severe exercise intolerance and multisystem manifestations. In this study, we characterized the biochemical properties of cybrids carrying this mutation and report that the homoplasmic p.278Y>C mutation caused a dramatic reduction in the CIII activity and in CIII-driven mitochondrial ATP synthesis. However, the CI, CI 1 CIII and CII 1 CIII activities and the rate of ATP synthesis driven by the CI or CII substrate were only partially reduced or unaffected. Consistent with these findings, mutated cybrids maintained the mitochondrial membrane potential in the presence of oligomycin, indicating that it originated from the respiratory electron transport chain. The p.278Y>C mutation enhanced superoxide production, as indicated by direct measurements in mitochondria and by the imbalance of glutathione homeostasis in intact cybrids. Remarkably, although the assembly of CI or CIII was not affected, the examination of respiratory supercomplexes revealed that the amounts of CIII dimer and III2IV1 were reduced, whereas those of I1III2IVn slightly increased. We therefore suggest that the deleterious effects of p.278Y>C mutation on cytochrome b are palliated when CIII is assembled into the supercomplexes I1III2IVn, in contrast to when it is found alone. These findings underline the importance of supramolecular interactions between complexes for maintaining a basal respiratory chain activity and shed light to the molecular basis of disease manifestations associated with this mutation.

U2 - 10.1093/hmg/ddt067

DO - 10.1093/hmg/ddt067

M3 - Journal article

C2 - 23418307

SN - 0964-6906

VL - 22

SP - 2141

EP - 2151

JO - Human Molecular Genetics

JF - Human Molecular Genetics

IS - 11

ER -

The cytochrome b p.278Y>C mutation causative of a multisystem disorder enhances superoxide production and alters supramolecular interactions of respiratory chain complexes

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