Analysis of the Chloroplast Protein Kinase Stt7 during State Transitions

State transitions allow for the balancing of the light excitation energy between photosystem I and photosystem II and for optimal photosynthetic activity when photosynthetic organisms are subjected to changing light conditions. This process is regulated by the redox state of the plastoquinone pool through the Stt7/STN7 protein kinase required for phosphorylation of the light-harvesting complex LHCII and for the reversible displacement of the mobile LHCII between the photosystems. We show that Stt7 is associated with photosynthetic complexes including LHCII, photosystem I, and the cytochrome b6f complex. Our data reveal that Stt7 acts in catalytic amounts. We also provide evidence that Stt7 contains a transmembrane region that separates its catalytic kinase domain on the stromal side from its N-terminal end in the thylakoid lumen with two conserved Cys that are critical for its activity and state transitions. On the basis of these data, we propose that the activity of Stt7 is regulated through its transmembrane domain and that a disulfide bond between the two lumen Cys is essential for its activity. The high-light–induced reduction of this bond may occur through a transthylakoid thiol–reducing pathway driven by the ferredoxin-thioredoxin system which is also required for cytochrome b6f assembly and heme biogenesis

Photosynthetic growth despite a broken Q-cycle

Central in respiration or photosynthesis, the cytochrome bc1 and b6f complexes are regarded as functionally similar quinol oxidoreductases. They both catalyse a redox loop, the Q-cycle, which couples electron and proton transfer. This loop involves a bifurcated electron transfer step considered as being mechanistically mandatory, making the Q-cycle indispensable for growth. Attempts to falsify this paradigm in the case of cytochrome bc1 have failed. The rapid proteolytic degradation of b6f complexes bearing mutations aimed at hindering the Q-cycle has precluded so far the experimental assessment of this model in the photosynthetic chain. Here we combine mutations in Chlamydomonas that inactivate the redox loop but preserve high accumulation levels of b6f complexes. The oxidoreductase activity of these crippled complexes is sufficient to sustain photosynthetic growth, which demonstrates that the Q-cycle is dispensable for oxygenic photosynthesis

Gravité quantique à deux dimensions couplée à de la matière non-conforme

Finding a theory of quantum gravity describing in a consistent way the quantum properties of matter and spacetime geometry is one of the greatest challenges of modern theoretical physics. However after several decades of research, many conceptual and technical issues are still to be resolved. Insights on these questions can be given by simplified toy models that allow for exact computations. The first part of the thesis deals with two-dimensional quantum gravity. In two dimensions quantum gravity is much better understood and many computations can be carried out exactly. Whereas two-dimensional quantum gravity coupled to conformal matter has been widely studied and is now well understood, much less was known until recently when matter is non-conformal. First we compute the gravitational action for a massive scalar field on a Riemann surface with boundaries and then for a massive Majorana fermion on a manifold without boundary. The latter case corresponds to a CFT perturbed by a conformal perturbation and is usually tackled through the DDK ansatz, but the results do not seem to match. Finally we give a minisuperspace computation of the spectrum of the Mabuchi action, a functional that appears in the gravitational action for a massive scalar field. In the second part we focus on black hole thermal behaviour which provides a lot of insight of how a theory of quantum gravity should look like. In the context of string theory the AdS/CFT correspondence provides powerful tools for understanding the microscopic origin of black holes thermodynamics. We construct a quantum mechanical toy model based on holographic principles to study the dynamics of quantum black holes.Établir une théorie de gravité quantique qui décrit de manière cohérente les propriétés quantiques de la matière et de l'espace-temps est l'un des défis majeurs de la physique théorique. Malgré plusieurs décennies de recherches, de nombreux problèmes conceptuels et techniques doivent encore être résolus. L'étude de modèles simplifiés donne des idées de résolution. La première partie de la thèse traite de la gravité quantique bidimensionnelle. À deux dimensions, la gravité quantique est beaucoup mieux comprise et de nombreux calculs peuvent être faits exactement. Si la gravité quantique bidimensionnelle a été largement étudiée quand elle est couplée à de la matière conforme, le cas de la matière non-conforme était très peu connu jusque récemment. Nous calculons d'abord l'action gravitationnelle pour un champ scalaire massif sur une surface de Riemann avec bords puis pour un fermion de Majorana massif sur une variété compacte. Ce dernier cas correspond à une CFT perturbée par une perturbation conforme et est d'ordinaire étudié grâce à l'ansatz de DDK, mais les résultats sont différents. Finalement, on calcule le spectre de l'action de Mabuchi dans l'approximation du minisuperespace. La seconde partie étudie les propriétés thermales des trous noirs dans le contexte de la correspondance AdS/CFT. On construit un modèle de mécanique quantique fondé sur les principes holographiques pour simuler la dynamique des trous noirs quantiques. Ce modèle permet d'obtenir des résultats numériques exacts

Two-dimensional quantum gravity coupled to non-conformal matter

Établir une théorie de gravité quantique qui décrit de manière cohérente les propriétés quantiques de la matière et de l'espace-temps est l'un des défis majeurs de la physique théorique. Malgré plusieurs décennies de recherches, de nombreux problèmes conceptuels et techniques doivent encore être résolus. L'étude de modèles simplifiés donne des idées de résolution. La première partie de la thèse traite de la gravité quantique bidimensionnelle. À deux dimensions, la gravité quantique est beaucoup mieux comprise et de nombreux calculs peuvent être faits exactement. Si la gravité quantique bidimensionnelle a été largement étudiée quand elle est couplée à de la matière conforme, le cas de la matière non-conforme était très peu connu jusque récemment. Nous calculons d'abord l'action gravitationnelle pour un champ scalaire massif sur une surface de Riemann avec bords puis pour un fermion de Majorana massif sur une variété compacte. Ce dernier cas correspond à une CFT perturbée par une perturbation conforme et est d'ordinaire étudié grâce à l'ansatz de DDK, mais les résultats sont différents. Finalement, on calcule le spectre de l'action de Mabuchi dans l'approximation du minisuperespace. La seconde partie étudie les propriétés thermales des trous noirs dans le contexte de la correspondance AdS/CFT. On construit un modèle de mécanique quantique fondé sur les principes holographiques pour simuler la dynamique des trous noirs quantiques. Ce modèle permet d'obtenir des résultats numériques exacts.Finding a theory of quantum gravity describing in a consistent way the quantum properties of matter and spacetime geometry is one of the greatest challenges of modern theoretical physics. However after several decades of research, many conceptual and technical issues are still to be resolved. Insights on these questions can be given by simplified toy models that allow for exact computations. The first part of the thesis deals with two-dimensional quantum gravity. In two dimensions quantum gravity is much better understood and many computations can be carried out exactly. Whereas two-dimensional quantum gravity coupled to conformal matter has been widely studied and is now well understood, much less was known until recently when matter is non-conformal. First we compute the gravitational action for a massive scalar field on a Riemann surface with boundaries and then for a massive Majorana fermion on a manifold without boundary. The latter case corresponds to a CFT perturbed by a conformal perturbation and is usually tackled through the DDK ansatz, but the results do not seem to match. Finally we give a minisuperspace computation of the spectrum of the Mabuchi action, a functional that appears in the gravitational action for a massive scalar field. In the second part we focus on black hole thermal behaviour which provides a lot of insight of how a theory of quantum gravity should look like. In the context of string theory the AdS/CFT correspondence provides powerful tools for understanding the microscopic origin of black holes thermodynamics. We construct a quantum mechanical toy model based on holographic principles to study the dynamics of quantum black holes

The purified mechanosensitive channel TREK-1 is directly sensitive to membrane tension.

International audienceMechanosensitive channels are detected in all cells and are speculated to play a key role in many functions including osmoregulation, growth, hearing, balance, and touch. In prokaryotic cells, a direct gating of mechanosensitive channels by membrane tension was clearly demonstrated because the purified channels could be functionally reconstituted in a lipid bilayer. No such evidence has been presented yet in the case of mechanosensitive channels from animal cells. TREK-1, a two-pore domain K(+) channel, was the first animal mechanosensitive channel identified at the molecular level. It is the target of a large variety of agents such as volatile anesthetics, neuroprotective agents, and antidepressants. We have produced the mouse TREK-1 in yeast, purified it, and reconstituted the protein in giant liposomes amenable to patch clamp recording. The protein exhibited the expected electrophysiological properties in terms of kinetics, selectivity, and pharmacology. Negative pressure (suction) applied through the pipette had no effect on the channel, but positive pressure could completely and reversibly close the channel. Our interpretation of these data is that the intrinsic tension in the lipid bilayer is sufficient to maximally activate the channel, which can be closed upon modification of the tension. These results indicate that TREK-1 is directly sensitive to membrane tension