ATP is the universal fuel molecule of any cell. The F1Fo-ATP synthase is a key enzyme of the energetic metabolism since it is responsible of most of the cellular synthesis of ATP. This 600 kDa complex is composed in yeast by 17 distinct subunits. It uses the energy of an electrochemical proton gradient to synthesize ATP. When protons are conducted across its membrane region, this conduction drives the rotation of a rotor part. This rotor part protrudes in the hydrophilic catalytic sector and induces conformational changes that lead to ATP synthesis. The yeast enzyme, like mammalian ATP synthases, is not only involved in ATP synthesis but also in the organization of the inner mitochondrial membrane: ATP synthase dimers constitute the building blocks of large oligomers that are involved in mitochondrial cristae morphology. Three ATP synthase subunits are essential for ATP synthase dimerization: su g, su e and the N-terminal extremity of subunit 4 including its two transmembrane segments (Nter 4). Despite the tremendous structural work performed on this complex, only 3D structures of sub-complexes have been solved and structural data are still missing on the channel region, made by subunit 6 and a ring of 10 subunits 9 (9(10) ring). Hence proton conduction is still a mystery. Besides, no information has been obtained on the structure and the organization of the small hydrophobic subunits e, g and the N-terminal part of subunit 4 that create the interfaces for ATP synthase oligomerization. This project will be divided in two main modules: - The first one will aim to get structural data on the channel part of the ATP synthase complex. To this end, two approaches will be used either by crystallizing the whole enzyme in lipidic sponge phase or in bicelles or by using classical 3D and/or 2D crystallization methods to get crystals of an isolated sub-complex containing the proton channel of the enzyme (subunits i + 6 + 9(10)). - The second part of the project will focus on the small hydrophobic subunits involved in the ATP synthase dimerization. A production of these proteins, in the presence or not of isotopically labelled amino acids will be performed using different modes of the cell-free expression system. After having determined which mode of production and which detergents or lipids would be the most appropriate for each protein, NMR spectra will be acquired to get data on their structures and the dynamics of their interactions. In combination to this structural and dynamical NMR studies, we will locate each subunit inside the dimeric ATP synthase species. This will be achieved by generating first a precise 3D volume of the dimeric complex by cryo-EM and then by replacing these small subunits in the envelope using a specific labelling with nanogold particles. The NMR models of subunits involved in the ATP synthase dimerization and the X-ray models of the whole enzyme and/or the proton channel will then be fitted into the 3D volume of dimers to get a precise model of the dimeric ATP synthase. This project will combine a lot of varied and complementary techniques (2D, 3D crystallization, X-ray crystallography, NMR, Cryo-EM, membrane protein expression) that are mandatory if we now want to decipher the intricate mechanism of proton translocation and thus the energetic coupling of this fantastic nanomotor that is the ATP synthase. Getting structural data on the small subunits involved in the dimerization of this complex will help understanding how the basic building blocks essential to the mitochondrial morphology are stabilized.

In SW Turkey, the oldest Homo erectus remains were dated to ca 1.6 - 1.2 Ma Lebatard et al., 2014; Vialet et al., 2012), documenting the migratory axis of Hominins populations from Africa to Europe. In the same area, we have access to exceptional lacustrine archives at Acigöl and Çatal Hüyük, over 1500 m of sedimentary infilling with the base dated to 2.4 Ma (Acigöl) and > 3 Ma (Çatal Hüyük), and to palaeontological sites rich in megafauna. Never, on the continent, have ancient archives been preserved in such a context that they now allow us to study the biological, geological and palaeoanthropological components in order to reconstruct in a systemic and interdisciplinary approach: 1) the interrelations and feedback between the different actors responsible for the functioning of the environments in a dynamic geological context (tectonics, volcanism); 2) the major Quaternary climatic cycles to be replaced into regional and global climate records and models; and 3) the evolving ecosystems (vegetation, fauna, Hominins), as well as the climate of this key region of the Eastern Mediterranean. The identification of the pollen grain of cereals (including rye) and nowadays cultivated trees (olive, walnut, plane, chestnut, arborescent Rosaceae) in the serie of Acigöl, will enable us to examine in detail the age and the modalities of the premises of domestication in this important area of Eastern Mediterranean frequented by human populations and large mammal herds. In particular: which micro-evolutive processes and forcing in this region of the Middle East are at the origin of the hereditary physiological transformation of these plants? It is a provocative question that is currently unanswered. Our results will therefore be highly significant for the understanding of the hominization of Europe and the genesis of the Neolithic Revolution. We will extend our research to the Middle East (Iran), Central Asia and Western Mediterranean sites (France, Italy, Spain, Greece) that populations of Hominins and large mammal herds frequented during their “Out of Africa” migration from ~2 Ma (Fig.1). The question is to understand if the association between palaeocereals, big mammals and Hominins that were evidenced at Acigöl existed elsewhere; knowing that humans certainly did not migrate alone but followed the ecosystem which suited them. With our consortium (ACIGOL), we will use concepts and techniques from life sciences (biology, genetics), earth sciences (geology, geophysics, isotopic geochemistry) and humanities sciences (palaeontology, archaeology, and ethnology) to carry out our project. The scientific program is organized around three key tasks: 1) Physical changes of the landscape and chronology; 2) Ecosystems changes and relationships with human populations; 3) Integrated understanding of the environment, ecosystems, human societies and climate interrelations The results will be published in specialized journals, as well as in journals of general interest, with a high impact factor and widely disseminated internationally. This research project is both unique and ambitious; the findings will be highly significant in terms of providing new understanding of the complex dynamics between past climates, environments, megaherbivores and human societies, as well as providing valuable data relevant to future climate change scenarios. The realization of this project will only be possible with the establishment of a strong consortium and the substantial financial support provided by ERC SYNERGY calls.