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Mitochondrial respiratory-chain complexes from Euglenozoa comprise classical subunits described in other eukaryotes (i.e. mammals and fungi) and subunits that are restricted to Euglenozoa (e.g. Euglena gracilis and Trypanosoma brucei). Here we studied the mitochondrial F1FO-ATP synthase (or Complex V) from the photosynthetic eukaryote E. gracilis in detail. The enzyme was purified by a two-step chromatographic procedure and its subunit composition was resolved by a three-dimensional gel electrophoresis (BN/SDS/SDS). Twenty-two different subunits were identified by mass-spectrometry analyses among which the canonical α, β, γ, δ, ε, and OSCP subunits, and at least seven subunits previously found in Trypanosoma. The ADP/ATP carrier was also associated to the ATP synthase into a dimeric ATP synthasome. Single-particle analysis by transmission electron microscopy of the dimeric ATP synthase indicated that the structures of both the catalytic and central rotor parts are conserved while other structural features are original. These new features include a large membrane-spanning region joining the monomers, an external peripheral stalk and a structure that goes through the membrane and reaches the inter membrane space below the c-ring, the latter having not been reported for any mitochondrial F-ATPase.

Metabolic function and architecture of mitochondria are intimately linked. More than 60 years ago, cristae were discovered as characteristic elements of mitochondria that harbor the protein complexes of oxidative phosphorylation, but how cristae are formed, remained an open question. Here we present experimental results obtained with yeast that support a novel hypothesis on the existence of two molecular pathways that lead to the generation of lamellar and tubular cristae. Formation of lamellar cristae depends on the mitochondrial fusion machinery through a pathway that is required also for homeostasis of mitochondria and mitochondrial DNA. Tubular cristae are formed via invaginations of the inner boundary membrane by a pathway independent of the fusion machinery. Dimerization of the F1FO-ATP synthase and the presence of the MICOS complex are necessary for both pathways. The proposed hypothesis is suggested to apply also to higher eukaryotes, since the key components are conserved in structure and function throughout evolution. DOI: http://dx.doi.org/10.7554/eLife.18853.001, eLife digest Cells contain compartments called mitochondria, which are often called the powerhouses of the cell because they provide energy that drives vital cellular processes. Mitochondria have two membranes: an outer and an inner membrane. The outer membrane separates the mitochondria from the rest of the cell. The inner membrane is elaborately folded and the folds – called cristae – create a larger space to accommodate all of the protein machinery involved in producing energy. The cristae can be shaped as flat sac-like structures called lamellar cristae or as tubes known as tubular cristae. Mitochondria are dynamic and are constantly fusing with other mitochondria and splitting up. Even though the internal architecture of mitochondria was first revealed around 60 years ago, it is still not clear how the cristae form. Harner et al. now address this question in yeast cells by combining imaging, biochemistry and genetic approaches. The experiments show that lamellar cristae form when two mitochondria fuse with each other. The outer membranes merge and then the inner membranes start to fuse around their edges to generate the sac-like structure of lamellar cristae. A yeast protein called Mgm1 (known as Opa1 in mammals) drives the fusion of the inner membranes, but this process only takes place when enzymes called F1FO-ATP synthases on the inner membrane form pairs with one another. These F1FO-ATP synthase pairs stabilize the cristae membranes as they curve to form the sac-like structure. Later on, the formation of a group of proteins called the MICOS complex halts the fusion process to prevent the lamellar cristae from completely separating from the rest of the inner membrane. Harner et al. also found that tubular cristae form using a different mechanism when the inner membrane of the mitochondria grows inwards. This process also requires pairs of F1FO-ATP synthases and the MICOS complex, but does not involve Mgm1/Opa1. Together, these findings show that lamellar and tubular cristae in yeast form using two different mechanisms. Since the key components of these mechanisms are also found in virtually all other eukaryotes, the findings of Harner et al. are also likely to apply to many other organisms including animals. DOI: http://dx.doi.org/10.7554/eLife.18853.002