Expression of the Ciona intestinalis Alternative Oxidase (AOX) in Drosophila Complements Defects in Mitochondrial Oxidative Phosphorylation

Fernández-Ayala, Daniel J. M.; Sanz, Alberto; Vartiainen, Suvi; Kemppainen, Kia K.; Babušiak, Marek; Mustalahti, Eero; Costa, Rodolfo; Tuomela, Tea; Zeviani, Massimo; Chung, Jongkyeong; O’Dell, Kevin; Rustin, Pierre; Jacobs, Howard T.

doi:10.1016/j.cmet.2009.03.004

Cited by 164 publications

(250 citation statements)

References 60 publications

Supporting

Mentioning

239

Contrasting

Order By: Relevance

“…Yeast NDI1, amplified from genomic DNA, was cloned into a Drosophila P-element vector under the control of a GAL4-dependent promoter, flanked by insulator elements. Following microinjection (VANEDIS Drosophila Injection Service), transgenic progeny were established as independent lines NDI1 A46 and NDI1 B20 , with genomic insertion sites (on chromosomes 3 and 2, respectively) determined by inverse PCR (13). NDI1 A46 ∕3 and NDI1 B20 ∕2 denote these transgenes in combination with the corresponding wild-type chromosomes.…”

Section: Methodsmentioning

confidence: 99%

“…w 1118 , balancer, and da-GAL4 driver lines were obtained from stock centers, and maintained in standard medium (13). AOX transgenic lines were as described previously (13).…”

Section: Methodsmentioning

confidence: 99%

“…RNA was isolated and analyzed by quantitative RT-PCR as described previously (13), using gene-specific primers, with RpL32 as reference gene. Subcellular fractions were analyzed by Western blots (13) and blue native-polyacrylamide gel electrophoresis combined with in-gel histochemistry (47). Table S1.…”

Section: Methodsmentioning

confidence: 99%

“…Furthermore, their expression appears largely benign, and confers resistance against respiratory chain inhibition by toxins or functional gene knockdown. Specifically, expression of AOX from the sea-squirt Ciona intestinalis was able to protect both flies (13) and mammalian cells (11) from the toxicity of cyanide or antimycin, or from functional depletion of cytochrome oxidase (12,13). Similarly, yeast Ndi1 was able to protect rat neurons from rotenone toxicity in vivo (15), and functionally to replace complex I in both Caenorhabditis elegans (14) and cultured human cells (8)(9)(10)).…”

mentioning

confidence: 96%

“…In previous studies these bypass enzymes were shown to be active when introduced into the mitochondria of higher metazoans such as mammals (8)(9)(10)(11)(12), arthropods (13), or nematodes (14), all of which lack endogenous alternative enzymes. Furthermore, their expression appears largely benign, and confers resistance against respiratory chain inhibition by toxins or functional gene knockdown.…”

mentioning

confidence: 99%

See 4 more Smart Citations

Expression of the yeast NADH dehydrogenase Ndi1 in Drosophila confers increased lifespan independently of dietary restriction

Sanz

Soikkeli²,

Portero‐Otín³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

125

162

View full text Add to dashboard Cite

Mutations in mitochondrial oxidative phosphorylation complex I are associated with multiple pathologies, and complex I has been proposed as a crucial regulator of animal longevity. In yeast, the single-subunit NADH dehydrogenase Ndi1 serves as a non-proton-translocating alternative enzyme that replaces complex I, bringing about the reoxidation of intramitochondrial NADH. We have created transgenic strains of Drosophila that express yeast NDI1 ubiquitously. Mitochondrial extracts from NDI1-expressing flies displayed a rotenone-insensitive NADH dehydrogenase activity, and functionality of the enzyme in vivo was confirmed by the rescue of lethality resulting from RNAi knockdown of complex I. NDI1 expression increased median, mean, and maximum lifespan independently of dietary restriction, and with no change in sirtuin activity. NDI1 expression mitigated the aging associated decline in respiratory capacity and the accompanying increase in mitochondrial reactive oxygen species production, and resulted in decreased accumulation of markers of oxidative damage in aged flies. Our results support a central role of mitochondrial oxidative phosphorylation complex I in influencing longevity via oxidative stress, independently of pathways connected to nutrition and growth signaling.aging | mitochondria | respiratory chain | free radicals M itochondria are key metabolic organelles whose oxidative phosphorylation (OXPHOS) system is considered to be one of the most efficient producers of bioenergy. When OX-PHOS function is compromized (e.g., by mutations or toxins), bioenergy supply and cellular homeostasis are seriously disrupted, which can be lethal.OXPHOS complex I plays a central role in the regulation of ATP production, intermediary metabolism, and apoptosis (1, 2), and mutations affecting it cause many human pathologies (3). It has also been proposed as a pacemaker of the aging process (4). Treatments inferred to decrease the production of reactive oxygen species (ROS) at the level of complex I can prolong lifespan in Drosophila (5). All these characteristics make it essential to understand better the role of complex I in vivo and its involvement in aging.Many organisms possess alternative enzymes that can bypass or replace the proton-translocating complexes of the mitochondrial respiratory chain. These include the alternative oxidases (AOX) and the NADH dehydrogenases of the Ndi and Nde families. Together these enzymes provide an alternative respiratory chain that potentially allows the maintenance of redox homeostasis and intermediary metabolism under conditions where flux through the "standard" respiratory chain is limited by high ATP levels, the action of toxins or other physiological restraints (6, 7). AOX acts as a bypass of complexes III and IV, whereas Nde or Ndi can bypass complex I.In previous studies these bypass enzymes were shown to be active when introduced into the mitochondria of higher metazoans such as mammals (8-12), arthropods (13), or nematodes (14), all of which lack endogenous alternative enzymes. Fu...

show abstract

Section: Methodsmentioning

confidence: 99%

“…w 1118 , balancer, and da-GAL4 driver lines were obtained from stock centers, and maintained in standard medium (13). AOX transgenic lines were as described previously (13).…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 96%

mentioning

confidence: 99%

See 3 more Smart Citations

Expression of the yeast NADH dehydrogenase Ndi1 in Drosophila confers increased lifespan independently of dietary restriction

Sanz

Soikkeli²,

Portero‐Otín³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

125

162

View full text Add to dashboard Cite

show abstract

Xenotopic expression of alternative electron transport enzymes in animal mitochondria and their impact in health and disease

Camargo

Chioda

Rodrigues

et al. 2018

Cell Biology International

Self Cite

View full text Add to dashboard Cite

The mitochondrial respiratory chain in vertebrates and arthropods is different from that of most other eukaryotes because they lack alternative enzymes that provide electron transfer pathways additional to the oxidative phosphorylation (OXPHOS) system. However, the use of diverse experimental models, such as human cells in culture, Drosophila melanogaster and the mouse, has demonstrated that the transgenic expression of these alternative enzymes can impact positively many phenotypes associated with human mitochondrial and other cellular dysfunction, including those typically presented in complex IV deficiencies, Parkinson's, and Alzheimer's. In addition, these enzymes have recently provided extremely valuable data on how, when, and where reactive oxygen species, considered by many as “by‐products” of OXPHOS, can contribute to animal longevity. It has also been shown that the expression of the alternative enzymes is thermogenic in cultured cells, causes reproductive defects in flies, and enhances the deleterious phenotype of some mitochondrial disease models. Therefore, all the reported beneficial effects must be considered with caution, as these enzymes have been proposed to be deployed in putative gene therapies to treat human diseases. Here, we present a brief review of the scientific data accumulated over the past decade that show the benefits and the risks of introducing alternative branches of the electron transport into mammalian and insect mitochondria, and we provide a perspective on the future of this research field.

show abstract

Emerging therapeutic approaches to mitochondrial diseases

Wenz

Williams

Bacman

et al. 2010

Dev Disabil Res Revs

View full text Add to dashboard Cite

Mitochondrial diseases are very heterogeneous and can affect different tissues and organs. Moreover, they can be caused by genetic defects in either nuclear or mitochondrial DNA as well as by environmental factors. All of these factors have made the development of therapies difficult. In this review article, we will discuss emerging approaches to the therapy of mitochondrial disorders, some of which are targeted to specific conditions whereas others may be applicable to a more diverse group of patients.

show abstract

Expression of the Ciona intestinalis Alternative Oxidase (AOX) in Drosophila Complements Defects in Mitochondrial Oxidative Phosphorylation

Cited by 164 publications

References 60 publications

Expression of the yeast NADH dehydrogenase Ndi1 in Drosophila confers increased lifespan independently of dietary restriction

Expression of the yeast NADH dehydrogenase Ndi1 in Drosophila confers increased lifespan independently of dietary restriction

Xenotopic expression of alternative electron transport enzymes in animal mitochondria and their impact in health and disease

Emerging therapeutic approaches to mitochondrial diseases

Contact Info

Product

Resources

About