Consequences of the pathogenic T9176C mutation of human mitochondrial DNA on yeast mitochondrial ATP synthase

Several human neurological disorders have been associated with various mutations affecting mitochondrial enzymes involved in cellular ATP production. One of these mutations, T9176C in the mitochondrial DNA (mtDNA), changes a highly conserved leucine residue into proline at position 217 of the mitochondrially encoded Atp6p (or a) subunit of the F1FO-ATP synthase. The consequences of this mutation on the mitochondrial ATP synthase are still poorly defined. To gain insight into the primary pathogenic mechanisms induced by T9176C, we have investigated the consequences of this mutation on the ATP synthase of yeast where Atp6p is also encoded by the mtDNA. In vitro, yeast atp6-T9176C mitochondria showed a 30% decrease in the rate of ATP synthesis. When forcing the F1FO complex to work in the reverse mode, i.e. F1-catalyzed hydrolysis of ATP coupled to proton transport out of the mitochondrial matrix, the mutant showed a normal proton-pumping activity and this activity was fully sensitive to oligomycin, an inhibitor of the ATP synthase proton channel. However, under conditions of maximal ATP hydrolytic activity, using non-osmotically protected mitochondria, the mutant ATPase activity was less efficiently inhibited by oligomycin (60% inhibition versus 85% for the wild type control). Blue Native Polyacrylamide Gel Electrophoresis analyses revealed that atp6-T9176C yeast accumulated rather good levels of fully assembled ATP synthase complexes. However, a number of sub-complexes (F1, Atp9p-ring, unassembled alpha-F1 subunits) could be detected as well, presumably because of a decreased stability of Atp6p within the ATP synthase. Although the oxidative phosphorylation capacity was reduced in atp6-T9176C yeast, the number of ATP molecules synthesized per electron transferred to oxygen was similar compared with wild type yeast. It can therefore be inferred that the coupling efficiency within the ATP synthase was mostly unaffected and that the T9176C mutation did not increase the proton permeability of the mitochondrial inner membrane

Wild food plants of popular use in Sicily

In the present work the authors report the result of their food ethnobotanical researches, which have been carried out in Sicily during the last thirty years. Data concerning 188 wild species used in the traditional Sicilian cuisine are reported. The authors underline those species that are partially or completely unknown for their culinary use and they illustrate other species that local inhabitants suggested in the prevention or treatment of symptomatologies caused by a refined diet, poor in vegetables. These data want to contribute to avoid the loss of traditional knowledge on uses and recipes concerning wild food botanicals, and to encourage further studies for those species that have not yet been sufficiently researched in their food chemical and nutritional profile. These studies may also suggest new applications for a few botanicals in medico-nutritional fields. The work includes also a short review of the seaweeds and mushrooms traditionally gathered and consumed in Sicily

Wild vascular plants gathered for consumption in the Polish countryside: a review

BACKGROUND: This paper is an ethnobotanical review of wild edible plants gathered for consumption from the end of the 18(th )century to the present day, within the present borders of Poland. METHODS: 42 ethnographic and botanical sources documenting the culinary use of wild plants were analyzed. RESULTS: The use of 112 species (3.7% of the flora) has been recorded. Only half of them have been used since the 1960s. Three species: Cirsium rivulare, Euphorbia peplus and Scirpus sylvaticus have never before been reported as edible by ethnobotanical literature. The list of wild edible plants which are still commonly gathered includes only two green vegetables (Rumex acetosa leaves for soups and Oxalis acetosella as children's snack), 15 folk species of fruits and seeds (Crataegus spp., Corylus avellana, Fagus sylvatica, Fragaria vesca, Malus domestica, Prunus spinosa, Pyrus spp., Rosa canina, Rubus idaeus, Rubus sect. Rubus, Sambucus nigra, Vaccinium myrtillus, V. oxycoccos, V. uliginosum, V. vitis-idaea) and four taxa used for seasoning or as preservatives (Armoracia rusticana root and leaves, Carum carvi seeds, Juniperus communis pseudo-fruits and Quercus spp. leaves). The use of other species is either forgotten or very rare. In the past, several species were used for food in times of scarcity, most commonly Chenopodium album, Urtica dioica, U. urens, Elymus repens, Oxalis acetosella and Cirsium spp., but now the use of wild plants is mainly restricted to raw consumption or making juices, jams, wines and other preserves. The history of the gradual disappearance of the original barszcz, Heracleum sphondylium soup, from Polish cuisine has been researched in detail and two, previously unpublished, instances of its use in the 20(th )century have been found in the Carpathians. An increase in the culinary use of some wild plants due to media publications can be observed. CONCLUSION: Poland can be characterized as a country where the traditions of culinary use of wild plants became impoverished very early, compared to some parts of southern Europe. The present use of wild plants, even among the oldest generation, has been almost entirely restricted to fruits

Mitochondrial ATP synthase disorders: Molecular mechanisms and the quest for curative therapeutic approaches.

International audienc

An uncoupling protein homologue putatively involved in facultative muscle thermogenesis in birds.

The cDNA of an uncoupling protein (UCP) homologue was obtained by screening a chicken skeletal-muscle library. The predicted 307-amino-acid sequence of avian UCP (avUCP) is 55, 70, 70 and 46% identical with mammalian UCP1, UCP2 and UCP3 and plant UCP respectively. avUCP mRNA expression is restricted to skeletal muscle and its abundance was increased 1.3-fold in a chicken line showing diet-induced thermogenesis, and 3.6- and 2.6-fold in cold-acclimated and glucagon-treated ducklings developing muscle non-shivering thermogenesis respectively. The present data support the implication of avUCP in avian energy expenditure

Mitochondrial protein sorting as a therapeutic target for ATP synthase disorders

Mitochondrial diseases are systemic, prevalent and often fatal; yet treatments remain scarce. Identifying molecular intervention points that can be therapeutically targeted remains a major challenge, which we confronted via a screening assay we developed. Using yeast models of mitochondrial ATP synthase disorders, we screened a drug repurposing library, and applied genomic and biochemical techniques to identify pathways of interest. Here we demonstrate that modulating the sorting of nuclear-encoded proteins into mitochondria, mediated by the TIM23 complex, proves therapeutic in both yeast and patient-derived cells exhibiting ATP synthase deficiency. Targeting ​TIM23-dependent protein sorting improves an array of phenotypes associated with ATP synthase disorders, including biogenesis and activity of the oxidative phosphorylation machinery. Our study establishes mitochondrial protein sorting as an intervention point for ATP synthase disorders, and because of the central role of this pathway in mitochondrial biogenesis, it holds broad value for the treatment of mitochondrial diseases

A yeast-based assay identifies drugs active against human mitochondrial disorders

Due to the lack of relevant animal models, development of effective treatments for human mitochondrial diseases has been limited. Here we establish a rapid, yeast-based assay to screen for drugs active against human inherited mitochondrial diseases affecting ATP synthase, in particular NARP (neuropathy, ataxia, and retinitis pigmentosa) syndrome. This method is based on the conservation of mitochondrial function from yeast to human, on the unique ability of yeast to survive without production of ATP by oxidative phosphorylation, and on the amenability of the yeast mitochondrial genome to site-directed mutagenesis. Our method identifies chlorhexidine by screening a chemical library and oleate through a candidate approach. We show that these molecules rescue a number of phenotypes resulting from mutations affecting ATP synthase in yeast. These compounds are also active on human cybrid cells derived from NARP patients. These results validate our method as an effective high-throughput screening approach to identify drugs active in the treatment of human ATP synthase disorders and suggest that this type of method could be applied to other mitochondrial diseases