Ce travail de thèse concerne une transversale complète des Pyrénées centro-occidentales, où on a combiné la thermochronologie basse température (traces de fission sur apatites, TFA) avec une analyse structurale détaillée pour décrire les mouvements verticaux associés à l'évolution du système chevauchant, et pour déterminer l'influence de ce dernier sur le cycle sédimentation/enfouissement/exhumation des dépôts synorogéniques du bassin d'avant-chaine sud (bassins de Jaca et Ainsa). L'analyse TFA complète les données déjà publiées dans la Zone Axiale et la Zone Nord-Pyrénéenne, et constitue la première étude de ce genre dans un bassin d'avant-chaîne pyrénéen. Les données TFA sur la transversale du bassin sud-pyrénéen montrent une diminution vers le sud du degré d'effacement des traces de fission, traduisant la diminution vers le sud de la quantité d'enfouissement, supérieure à 5 km au nord et inférieure à 3 km au sud dans l'hypothèse un géotherme de 25°.km-1. Le contexte géologique montre que l'enfouissement est principalement lié à l'accumulation des dépôts synorogéniques. Les données TFA de la partie nord du bassin montrent un refroidissement d'âge Oligocène supérieur-Miocène inferieur (moyen). Par ailleurs, une nouvelle interprétation de profils de sismiques réflexion dans le bassin de Jaca montre que le chevauchement d'Oturia s'enracine dans le chevauchement de socle de Bielsa, responsable de l'exhumation tectonique hors-séquence du bord sud de la Zone Axiale au Miocène inférieur (-moyen) (Jolivet et al., 2007). Ces résultats attestent donc de l'exhumation tectonique hors-séquence au Miocène inférieur (Burdigalien- ?Langhien) de la partie nord du bassin d'avant-chaine sud-pyrénéen. Des données TFA obtenues dans la Zone Axiale et la Zone Nord-Pyrénéenne confirment la migration générale vers le sud du système chevauchant, et mettent également en évidence la réactivation tectonique hors-séquence du bord nord de la Zone Axiale à l'Oligocène terminal-Miocène inférieur. L'ensemble de ces résultats atteste donc de la réactivation en « pop-up » de la parties interne des Pyrénées centre-ouest à l'Oligocène supérieur-Miocène inférieur (Burdigalien- ?Langhien), postérieurement au scellement du front sud-pyrénéen (Aquitanien- ?Burdigalien) classiquement considéré comme marquant la fin de la compression pyrénéenne. Ces données nous ont permis de proposer un nouveau modèle d'évolution crustale des Pyrénées centro-occidentales en 3 grandes étapes : (i) du Crétacé supérieur à l'Eocène moyen, le prisme est caractérisé par une absence de relief, en lien avec l'inversion de structures extensives crétacées conduisant à l'accrétion de petites écailles crustales ; (ii) la période Eocène supérieur-Oligocène correspond à la collision continentale proprement dite, et est marquée par la création d'importants reliefs associés à l'accrétion d'épaisses unités crustales ; (iii) au Miocène inférieur, la partie interne du prisme pyrénéen est réactivée. / In this work on a complete transect of the west-central Pyrenees, we combine low temperature thermochronology (apatite fission tracks, AFT) with a detailed structural analysis to describe vertical movements related to the thrusting system evolution, and to determine the influence of the latter on the sedimentation/burial/exhumation cycle of the synorogenic deposits of the southern foreland basin (Jaca and Ainsa basins). AFT analysis from a transect of the south-Pyrenean basin show the southward decrease of the fission track reset level from the southern edge of the Axial Zone to the South-Pyrenean frontal thrust, implying the southwards decrease of the burial amount from more than 5km in the north to less than 3km in the south assuming an average geothermal gradient of 25°C.km-1. The structural setting of the Jaca basin attests that the burial of the synorogenic sediments was mainly due to the sedimentary accumulation. AFT data from the northern part of the basin display a late Oligocene-early (middle) Miocene cooling event. New interpretation of industrial seismic reflection profiles across the Jaca basin suggests that the Oturia thrust is rooted in the Bielsa basement thrust, responsible for the early (-middle) Miocene out-of-sequence tectonic reactivation of the southern flank of the Axial Zone (Jolivet et al., 2007). These results reveal a lower Miocene (Burdigalian -?Langhian) out-of-sequence episode of tectonic activity of the interior of the south-Pyrenean foreland basin. AFT data from the Axial Zone and the North-Pyrenean Zone confirm the general southward migration of the thrusting system, and also bring evidence of the late Oligocene-lower Miocene out-of-sequence tectonic reactivation of the northern flank of the Axial Zone. All these results attest of a late Oligocene-lower Miocene (Burdigalian-?Langhian) 'pop-up' reactivation of the inner part of the west-central Pyrenees, younger than the sealing of the south-Pyrenean front (Aquitanian-?Burdigalian) which is classically considered to mark the end of the Pyrenean compression. These results lead us to propose a new crustal scale evolution model of the west-central Pyrenees in 3 stages: (i) From the Late Cretaceous to the middle Eocene, the orogenic prism is characterised by the absence of relief, related to the inversion of Cretaceous extensional structures leading to the accretion of thin crustal units; (ii) The late Eocene-Oligocene stage corresponds to the continental collision, marke d by the creation of important relief associated with the accretion of thick crustal units; (iii) During the early Miocene, the inner part of the Pyrenean wedge is tectonically reactivated.

L'isthme de Panama se situe en bordure SW de la plaque Caraïbe, à la jonction de trois plaques lithosphériques: les plaques Amérique du Sud, Nazca et Cocos. Cet isthme est constitué de deux arcs volcaniques formant l'Amérique Centrale. Leurs présences reflètent une histoire complexe de convergence, en subduction. L'événement majeur de cette région correspond à la collision de l'Amérique Centrale contre l'Amérique du Sud entre 12-25 Ma. L'objectif de cette thèse est de documenter les déformations avant, pendant et après le processus d'accrétion continentale. Le but est de mieux comprendre comment un arc volcanique s'accrète sur une marge continentale pour reconstruire l'histoire géodynamique de cette région de 70 Ma jusqu'à nos jours. Cette thèse combine: - une étude sédimentologique et paléontologique, - une étude structurale à partir de données spatiales, géophysiques, et de terrain, - une étude thermochronologique (AFT), - et une étude interprétative sismique. Je propose ainsi une accrétion progressive et oblique de l'Amérique Centrale sur l'Amérique du Sud, s'initiant au sud de la région d'Istmina à partir de 40-37 Ma. La plaque Caraïbe, piégée entre l'arc volcanique et la marge continentale sud-américaine, disparaît progressivement sous l'Amérique du Sud. Vers 15 Ma, l'accrétion de l'arc dans la partie colombienne se termine. Au Panama, la convergence continentale se poursuit, mais le système s'inverse. Une nouvelle subduction s'initie : la plaque Caraïbe subducte sous l'isthme. Les déformations compressives engendrées par l'accrétion contrôlent la migration des masses sédimentaires et permettent la surrection progressive de l'isthme créant le pont inter-Amériques. / The Panama Isthmus is located on the SW boundary of the Caribbean plate, at the junction of the South American, Nazca and Cocos plates. The isthmus is composed of two island arcs forming Central America. It formed by a complex history of plate subductions. The major tectonic event in this region is attributed to the accretion of Central America with South America between 12 and 25 Ma. The aim of this thesis is to document the deformation before, during and after the accretionary continental process. The main purpose is to better understand how a volcanic arc collides against a continental margin in order to reconstruct the tectonic history of this region since 70 Ma until today. This thesis combines: - a sedimentological and paleontological studies, - a structural study from spatial, geophysical and field work data, - a thermochronological study (AFT), - and an interpretative seismic study. I propose the initiation of progressive and oblique arc-continent collision during 40-37 Ma. The Caribbean plate, trapped between the arc and the continent, progressively disappeared beneath the South American continent. Around 15 Ma, the Colombian part of Central America was accreted and the convergence of Panama toward the continent progressed and produced a new subduction zone whereby the Caribbean plate subducted beneath the Panama Isthmus. Compressive deformations, caused by the collision, still actively control the migration of sedimentary masses, allowing the progressive emergence of the isthmus and forming the inter-American land bridge.

Comprendre les mécanismes de spéciation des Produits de Fission (PF) dans le combustible nucléaire est un enjeu majeur pour pouvoir estimer précisément le terme source d’un accident grave. Parmi les nombreux PF créés, certains sont très réactifs et peuvent avoir un impact radiologique important en cas de relâchement dans l’atmosphère. C’est notamment le cas du césium (Cs), du molybdène (Mo) et du baryum (Ba). C’est dans ce contexte que s’inscrit le travail de thèse qui propose d’apporter des données expérimentales sur l’effet du potentiel oxygène sur la spéciation du Cs, du Mo et du Ba dans des combustibles nucléaires, à différents stades d’un accident grave.Une approche thermodynamique a été utilisée en support à l’interprétation des données expérimentales obtenues dans le cadre de ce travail. Deux types d’échantillons ont été étudiés: des combustibles MOX irradiés et des matériaux simulant un combustible UO2 à fort taux de combustion, obtenus par frittage à haute température (SIMFuel). Les échantillons ont été traités thermiquement dans des conditions représentatives d’un accident grave survenant dans un Réacteur à Eau Pressurisée (REP). Les conditions expérimentales ont couvert une gamme de température allant de 400°C à 2530°C et des potentiels oxygène situés entre -470 kJ.mol(O2)-1 et -100 kJ.mol(O2)-1. Les échantillons ont été caractérisés finement avant et après chaque traitement à l’aide de techniques complémentaires comme la microscopie optique et électronique, la microsonde et le SIMS dans le cas de l’irradié. Des mesures de XANES sur synchrotron ont été réalisées sur SIMFuel et ont conduit à des résultats importants en termes de spéciation des PF. Enfin, la technique de Spark Plasma Sintering (SPS) a été explorée avec succès pour la fabrication de SIMFuel contenant du Cs, du Mo et du Ba sous des formes chimiques représentatives d’un combustible REP en fonctionnement nominal.Ce travail a permis de mettre en évidence l’effet de la température en conditions oxydantes sur le comportement du combustible et des PF. Une oxydation du Mo, initialement présent sous forme métallique dans les inclusions blanches du combustible, en MoO2 a été observée dès 1000°C en conditions oxydantes. Une interaction entre le MoO2 formé et le Ba contenu dans la phase oxyde a eu lieu dans les mêmes conditions, menant à la formation de BaMoO4. Le potentiel oxygène joue aussi un rôle important dans le phénomène d’interaction pastille-gaine, en favorisant la diffusion des espèces en conditions oxydantes, diminuant ainsi la température de fusion du combustible. / In the nuclear community, it is a top priority to gain in-depth understanding of fission product (FP) speciation mechanisms occurring in nuclear fuel in order to precisely estimate the source term of a severe accident. Among the FP produced, some are highly reactive and may have a strong radiological impact if released into the environment. This is particularly the case of cesium (Cs), molybdenum (Mo) and barium (Ba). In this context, the objective of this study is to provide experimental data on the effect of the oxygen potential on Cs, Mo and Ba speciation in nuclear fuels at different stages of a severe accident.A thermodynamic approach was coupled with the experimental work to support the interpretation of experimental data. Two types of samples were studied in detail: irradiated MOX fuels and simulated high burn-up UO2 fuels produced through sintering at high temperature (SIMFuel). The samples were submitted to thermal treatments in conditions representative of a pressurised water reactor (PWR) severe accident. This approach made it possible to cover a temperature range from 400°C up to 2530°C and oxygen potentials from -470 kJ.mol(O2)-1 to -100 kJ.mol(O2)-1. The samples were characterized before and after each test using complementary techniques like OM, SEM, EPMA and SIMS in the case of irradiated fuels. XANES measurements using synchrotron radiation facilities were performed on SIMFuels and provided valuable results on FP speciation. Moreover, spark plasma sintering (SPS) was successfully investigated for the production of SIMFuel samples containing Cs, Mo and Ba in a chemical state representative of PWR fuel in normal operating conditions.This work highlighted the effect of oxidizing severe accident conditions on the fuel and FP behavior. Oxidation of Mo initially contained in the fuel’s metallic inclusions into MoO2 was observed to take place around 1000°C in oxidizing conditions. An interaction between MoO2 and the oxide phase containing Ba took place in the same conditions, leading to the formation of BaMoO4. The oxygen potential also plays an important role in fuel-cladding interactions, enhancing the diffusion of species in oxidizing conditions and lowering the temperature at which fuel melting occurs.

Le but de ces travaux de thèse est d’apporter une meilleure compréhension des mécanismes de régulation contrôlant la division cellulaire au niveau moléculaire. La division cellulaire est composée de la mitose et la cytocinèse. Les deux processus doivent être coordonnés étroitement afin de garantir la stabilité du génome. La division cellulaire doit aussi s’équilibrer avec la croissance cellulaire pour que les cellules conservent une taille constante au cours des cycles successifs. La levure S. pombe est un organisme modèle simple très utilisé pour des études de cycle cellulaire et de cytocinèse. Dans ce modèle, nous avons focalisé ce travail de thèse sur les nœuds corticaux médians, des structures protéiques complexes, qui ont une fonction double dans l’engagement en mitose et dans le positionnement du plan de division. Les nœuds médians corticaux sont organisés par la kinase SAD Cdr2. Leur localisation et leur fonction sont régulées négativement pour la DYRK kinase Pom1 qui forme des gradients émanant des extrémités de la cellule. Les nœuds corticaux médians contiennent une voie d’inhibition pour Wee1 qui promeut l’entrée en mitose. Cette voie implique la kinase SAD Cdr1, un inhibiteur direct de Wee1 et pourrait coupler l’entrée en mitose à la taille de la cellule par levée progressive de l’inhibition exercée par Pom1 quand les cellules s’allongent. Cdr2 recrute aussi l’anillin Mid1 sur les nœuds corticaux médians ainsi qu’une série de composants additionnels, Blt1, Gef2, Nod1 et Klp8, pour former des précurseurs médians de l’anneau contractile de cytocinèse qui se compactent en un anneau fin pendant la mitose. La localisation médiane des nœuds, contrôlée négativement par les gradients polaires de Pom1 prédéfinit ainsi le plan de division au centre géométrique de la cellule. Dans la première partie de ma thèse, j’ai étudié la protéine des nœuds corticaux médians Blt1 dont la fonction restait énigmatique. Nous avons montré que Blt1 promeut une association robuste de Mid1 avec les nœuds corticaux. Blt1 interagit avec Mid1 via le RhoGEF Gef2 pour stabiliser les nœuds au cortex cellulaire durant les premiers stades de l’assemblage de l’anneau contractile. L’extrémité N-terminale de Blt1 est nécessaire à sa localisation ainsi qu’à sa fonction, tandis que son extrémité C-terminale favorise sa localisation au cortex en interagissant avec des phospholipides. Dans des cellules dans lesquelles ni Mid1 ni Blt1 ne peuvent s’attacher à la membrane, les nœuds se détachent du cortex et génèrent des anneaux contractiles de cytocinèse aberrants. Nous en avons conclu que Blt1 agit comme une protéine d’échafaudage pour les précurseurs de l’anneau contractile, et que Blt1 et Mid1 constituent des ancres membranaires redondantes pour le positionnement du plan de division. Dans une deuxième partie de ma thèse, j’ai étudié comment Cdr2 organise les différents composants des nœuds en voies fonctionnelles qui favorisent l’entrée en mitose et la division médiane. J’ai montré que l’interaction de Cdr2 avec Wee1 et Mid1 dépend du domaine UBA de Cdr2 de manière dépendante de l’activité kinase. En revanche, Cdr1 s’associe avec l’extrémité C-terminale de Cdr2, composée des domaines basique et KA1 d’association aux lipides membranaires. De manière intéressante, Mid1 interagit également avec l’extrémité C-terminale de Cdr2 et pourrait ponter les parties N- et C-terminales de Cdr2, alors que Blt1 s’associe à la région centrale de Cdr2. Nous faisons l’hypothèse que l’association des effecteurs de Cdr2 avec différents domaines de Cdr2 pourraient contraindre Cdr1 et Wee1 spatialement pour promouvoir l'inhibition de Wee1 quand la kinase Cdr2 est active. / The aim of this PhD work is to bring a better understanding of the regulatory mechanism controlling cell division in space and time at the molecular level. Cell division is composed of mitosis and cytokinesis. Both processes need to be perfectly coordinated in order to guarantee genome integrity. Cell division also needs to be properly balanced with cell growth to maintain cell size constant during successive cell cycles. Temporal and spatial regulatory mechanisms ensure the coordination of these events. The fission yeast Schizosaccharomyces pombe is a simple rod-shaped model organism well-known for cell cycle and cytokinesis studies. In this model, we focused the work of this thesis on the medial cortical nodes, complexe protein structures that have a dual role in mitotic commitment and in division plane positioning. Medial cortical nodes are organized by the SAD kinase Cdr2. Their localization and function is negatively regulated by the DYRK kinase Pom1 that forms a gradient emanating from the cell tips. Medial cortical nodes contain an inhibitory pathway for Wee1, promoting mitotic entry. This pathway involves the SAD kinase Cdr1, a direct inhibitor of Wee1 and has been proposed to couple mitotic entry to cell size by progressive alleviation of Pom1 inhibition when cells grow longer. Cdr2 also recruits to medial nodes the anillin Mid1 as well as a series of four additional components, Blt1, Gef2, Nod1 and Klp8, to form medial precursors for the cytokinetic contractile ring that compact into a tight ring during mitosis. Nodes medial localization, negatively controlled by Pom1 gradients, predefines thereby the division plane in the cell geometrical center. In a first part of my thesis, I studied the previously enigmatic cortical node protein Blt1. We showed that Blt1 promotes the robust association of Mid1 with cortical nodes. Blt1 interacts with Mid1 through the RhoGEF Gef2 to stabilize nodes at the cell cortex during the early stages of contractile ring assembly. The Blt1 N terminus is required for localization and function, while the Blt1 C terminus promotes cortical localization by interacting with phospholipids. In cells lacking membrane binding by both Mid1 and Blt1, nodes detach from the cell cortex and generate aberrant cytokinetic rings. We conclude that Blt1 acts as a scaffolding protein for precursors of the cytokinetic ring and that Blt1 and Mid1 provide overlapping membrane anchors for proper division plane positioning. In the second part of my thesis, I studied how Cdr2 scaffolds various nodes components to organize them in functional pathways promoting mitotic commitment and medial division. I showed that Cdr2 interaction with Wee1 and Mid1, depends on Cdr2 UBA domain in a kinase activity dependent manner. In contrast, Cdr1 associates with Cdr2 C-terminus composed of basic and KA-1 lipid-binding domains. Interestingly, Mid1 also interacts with Cdr2 C-terminus and may the bridge N- and C-terminal domains of Cdr2 while Blt1 associates with the central spacer region. We propose that the association of Cdr2 effectors with different Cdr2 domains may constrain Cdr1 and Wee1 spatially to promote Wee1 inhibition upon Cdr2 kinase activation.

Les sections efficaces induites par neutrons des noyaux de courte durée de vie jouent un rôle important dans des domaines variés parmi la physique fondamentale, l'astrophysique ou l'énergie nucléaire. Malheureusement de nombreuses contraintes liées à la radiotoxicité des cibles rendent la mesure de ces sections efficaces souvent très difficiles. La méthode de substitution est une technique de mesure indirecte de sections efficaces neutroniques de noyaux radioactifs qui à l'avantage de s'affranchir de ces contraintes. Pour la première fois dans ce type d'expérience,les probabilités de fission et d'émission gamma sont mesurées simultanément, pour les actinides236U, 238U, 237Np et 238Np dans le but d'étudier la validité de la méthode. Une des difficultés provient de la soustraction des gammas des fragments de fission et cette mesure constitue en cela un véritable défi. Cette expérience de mesure simultanée a été effectuée au cyclotron d'Oslo.A une énergie d'excitation fixée du noyau formé, les résultats montrent que les probabilités de fission de substitution sont en bon accord avec celles induites par neutron alors que les probabilités d'émission gamma mesurées sont plusieurs fois plus élevées. Ces écarts sont liés à la différence distribution spin peuplée par le noyau entre les deux méthodes. Des calculs de modèles statistiques avec des paramètres standards n'ont pas permis de reproduire cette insensibilité de la probabilité de fission vis à vis du spin du noyau. La reproduction des observations expérimentales devient possible en considérant un moment d'inertie du noyau fissionnant qui augmente plus rapidement avec la déformation du noyau que ne le préconisent les paramètres standards. De nouveaux efforts théoriques sont à fournir pour améliorer la compréhension de nos résultats. / Neutron-induced cross sections of short-lived nuclei are important in various fields such as fundamental physics, astrophysics or nuclear energy. However, these cross sections are often extremely difficult to measure due to high radioactivity of the targets involved. The surrogate-reaction method is an indirect way to determine neutron-induced cross sections of short-lived nuclei. In order to study the validity of the method, we have measured for the very first time in a surrogate-reaction experiment simultaneously fission and gamma-decay probabilities for the actinides 236U, 238U, 237Np and 238Np. This is challenging because one has to remove the gamma rays emitted by the fission fragments. The measurement was performed at the Oslocyclotron.Our results show that for a given excitation energy, our gamma-decay probabilities are several times higher than neutron-induced probabilities, which can be attributed to differences in spin distribution between the two types of reactions. On the other hand, our fission probabilities are in good agreement with neutron-induced data. Statistical-model calculations applied with standardparameters cannot reproduce the weak spin sensibility to variations of the angular momentum observed for the fission probabilities. However, it is possible to reproduce the experimental observations by considering a stronger increase of the moment of inertia of the fissionning nucleus with deformation. Further theoretical efforts are needed to improve the understanding of our results

Les travaux présentés visent à améliorer les modèles de physique nucléaireutilisés dans l’évaluation des sections efficaces neutroniques de fission. Le résultat deces travaux donne les clefs pour une percée significative dans ce domaine et a permisd’étendre fortement les capacités du code d’évaluation CONRAD. Les sections partiellesétant naturellement corrélées entre-elles pour respecter la valeur de la section totale, cesaméliorations bénéficient à l’ensemble des sections partielles. Un cadre solide pour lamodélisation des processus concurrent à la fission a dû être établi sur le modèle du codede référence TALYS. Après s’être assuré de la fiabilité et de la cohérence du cadre, lesinvestigations spécifiques concernant la fission ont pu être réalisées. Les perspectivesd’applications offertes par les modèles macro-microscopiques FRDM et FRLDM ont étéanalysées. Ces modèles ont été implémentés et validés sur des données expérimentaleset des benchmarks. Afin d’obtenir des temps de calcul compatibles avec les besoins del’évaluation, des méthodes numériques sophistiquées ont été sélectionnées et une partiedes calculs a été portée sur GPU. Ces modèles macro-microscopiques peuvent être utiliséspour construire des surfaces d’énergie potentielle qui sont à leur tour traitées afin d’obtenirdes barrières de fission à une dimension, puis des coefficients de transmission fission. Cesderniers sont alors utilisés dans le cadre de modélisation des sections efficaces moyennesdu domaine statistique sur la base d’un modèle Hauser-Feshbach. Les résultats de cetteapproche seront présentés sur le cas du 239Pu(n,f). / The work presented here aims to improve models used in the fission crosssectionevaluation. The results give insights for a significant breakthrough in this fieldand yielded large extensions of the evaluation code CONRAD. Partial cross sections areinherently strongly correlated together as of the competition of the related reactions mustyield the total cross section. Therefore improving fission cross section benefits to all partialcross sections. A sound framework for the simulation of competitive reactions hadto be settled in order to further investigate on the fission reaction; this was implementedusing the TALYS reference code as guideline. After ensuring consistency and consistencyof the framework, focus was made on fission. Perspective resulting from the useof macroscopic-microscopic models such as the FRDM and FRLDM were analyzed; thesemodels have been implemented and validated on experimental data and benchmarks. Tocomply with evaluation requirements in terms of computation time, several specific numericalmethods have been used and parts of the program were written to run on GPU.These macroscopic-microscopic models yield potential energy surfaces that can be used toextract a one-dimensional fission barrier. This latter can then be used to obtained fissiontransmission coefficients that can be used in a Hauser-Feshbach model. This method hasbeen finally tested for the calculation of the average fission cross section for 239Pu(n,f).

It is important to have a method of experimentally calculating fission product yields. Statistical calculations and simulations produce very large uncertainties. Experimental calculations, depending on the methods used, tend to produce lower uncertainties. This work set up a method to calculate fission product yields using gamma ray spectroscopy. In order to produce a method that was theoretically sound, a simulation was set up using OrigenArp to calculate theoretical concentrations of fission products from the irradiation of natural uranium. From these concentrations, the fission product yields were calculated to verify that they would agree with expected values. Moving forward in the work, the total flux at the point of irradiation, in the pneumatic transfer system, was calculated and determined to be 3.9070E+11 ± 6.9570E+10 n/cm^2/s at 100 kW. Once the flux was calculated, the method for calculating fission product yields was implemented and yields were calculated for 10 fission products. The yields calculated were in very good agreement (within 10.04%) with expected values taken from the ENDF-349 library. This method has strong potential in nuclear forensics as it can provide a means for developing a library of experimentally-determined fission product yields, as well as rapid post-nuclear detonation analysis. / text

Les fluorures fondus sont utilisés dans les Réacteurs à Sels Fondus tels que le réacteur non modéré, à neutrons rapides, MSFR où le sel fondu LiF-ThF4 joue le rôle du combustible et du liquide caloporteur. La formation des produits de fissions (PF) tels que les lanthanides, au cours du fonctionnement de ce réacteur, peut modifier les propriétés physicochimiques du bain fondu. Il est ainsi important de caractériser le bain fondu de point de vue structural et dynamique afin de remonter à ses propriétés. En raison des problèmes de radioactivité liés au thorium, et des conditions requises liées aux méthodes spectroscopiques utilisées, le système étudié dans ce manuscrit est le LiF-ZrF4-LaF3 (le zirconium et le lanthane étant des PF potentiels). L'approche développée dans cette thèse combine des mesures par spectroscopies RMN et EXAFS à 850 °C avec des simulations de dynamique moléculaire. Dans le bain fondu, nous avons montré la coexistence de complexes à base de zirconium et de lanthane de différentes coordinences, dont les proportions et les interactions dépendent de la composition. En fonction de la teneur en ZrF4, les espèces [ZrF7]3- majoritaires évoluent peu mais se connectent davantage via des fluors pontants. L’ajout de LaF3 au mélange stabilise la coordinence 7 autour du zirconium et tend à enrichir l’environnement du lanthane en fluors. Un ordre à moyenne portée s’établit entre les différents complexes à base de zirconium et de lanthane par l’intermédiaire des fluors pontants. La dynamique des espèces est ralentie en fonction de l’ajout de ZrF4 et LaF3. Nous avons noté un effet important sur la structure et la dynamique des espèces à partir de 10% mol. LaF3 ajouté au mélange. Les données obtenues par cette approche originale de la chimie du bain fondu dans le RSF en présence des PF, sont fondamentales pour améliorer la séparation de ces derniers et optimiser le procédé. / Molten fluorides are used in Molten Salt Reactors MSR such as the non moderated fast reactor MSFR, where the molten salt LiF-ThF4 is the fuel and the coolant. The formation of fission products (FP) such as lanthanides, during the reactor operation, possibly modifies the physicochemical properties of the melt. It is therefore important to characterize the melt from the structural and the dynamics point of view in order to determine its properties. Because of problems related to the radioactivity of thorium, as well as requirements related to spectroscopic methods, the system studied in this thesis is the LiF-ZrF4-LaF3 (zirconium and lanthanum are possible FP). The approach followed in this thesis combines measurements by NMR spectroscopy and EXAFS at 850 °C with molecular dynamics simulations. In the molten salt, we have shown the existence of zirconium and lanthanum complexes with different coordination numbers, whose proportions depend on the composition. Depending on the content of ZrF4, [ZrF7]3- species are dominant but change slightly and are further connected between each other’s via bridging fluorine. The addition of LaF3 to the mixture stabilizes the 7 coordination number around the zirconium and tends to enrich the environment of lanthanum with fluorides. A medium-range order is established between the various complexes containing zirconium and lanthanum due to bridging fluorine. Species dynamics is slower when the amount of either ZrF4 or LaF3 is higher. We noted a significant effect on the structure and dynamics of species starting 10 mol%. LaF3 added to the medium. The data obtained by this novel approach concerning the chemistry of the molten salt in MSR containing FP, are fundamental to improve the separation of these products and optimize the process.

Gene deletions are a powerful method to uncover the cellular functions of a given gene in living systems. A limitation to this methodology is that it is not applicable to essential genes. Even for non-essential genes, gene knockouts cause complete absence of gene product thereby limiting genetic analysis of the biological pathway. Alternatives to gene deletions are mutants that are conditional, for e.g, temperature sensitive (ts) mutants are robust tools to understand temporal and spatial functions of genes. By definition, products of such mutants have near normal activity at a lower temperature or near-optimal growth temperature which is called as the permissive temperature and reduced activity at a higher, non-optimal temperature called as the non-permissive temperature. Generation of ts alleles in genes of interest is often time consuming as it requires screening a large population of mutants to identify those that are conditional. Often many essential proteins do not yield ts such alleles even after saturation mutagenesis and extensive screening (Harris et al., 1992; Varadarajan et al., 1996). The limited availability of such mutants in many essential genes prompted us to adopt a biophysical approach to design temperature-sensitive missense mutants in an essential gene of fission yeast. Several studies report that mutations in buried or solvent-inaccessible amino acids cause extensive changes in the thermal stability of proteins and specific substitutions create temperature-sensitive mutants (Rennell et al., 1991; Sandberg et al., 1995). We used the above approach to generate conditional mutants in the fission yeast gene spprp18+encoding an essential predicted second splicing factor based on its homology with human and S. cerevisiae proteins. We have used a missense mutant coupled with a conditional expression system to elucidate the cellular functions of spprp18+. Further, we have employed the same biophysical principle to generate a missense mutant in spago1+ RNA silencing factor that is non-essential for viability but has critical functions in the RNAi pathway of fission yeast. Fission yeast pre-mRNA splicing: cellular functions for the protein factor SpPrp18 Pre-mRNA splicing is an evolutionarily conserved process that excises introns from nascent transcripts. Splicing reactions are catalyzed by the large ribonuclear protein machinery called the spliceosome and occur by two invariant trans-esterification reactions (reviewed in Ruby and Abelson, 1991; Moore et al., 1993). The RNA-RNA, RNA–protein and protein-protein interactions in an assembly of such a large protein complex are numerous and highly dynamic in nature. These interactions in in vitro splicing reactions show ordered recruitment of essential small nuclear ribonucleic particles snRNPs and non–snRNP components on pre-mRNA cis-elements. Further these trans acting factors recognize and poise the catalytic sites in proximity to identify and excise introns. The precision of the process is remarkable given the diversity in architecture for exons and introns in eukaryotic genes (reviewed in Burge et al., 1999; Will and Luhrmann, 2006). Many spliceosomal protein components are conserved across various organisms, yet introns have diverse features with large variations in primary sequence. We hypothesize that co-evolution of splicing factor functions occurs with changes in gene and intron architectures and argue for alternative spliceosomal interactions for spliceosomal proteins that thus enabling splicing of the divergent introns. In vitro biochemical and genetic studies in S. cerevisiae and biochemical studies with human cell lines have indicated that ScPRP18 and its human homolog hPRP18 function during the second catalytic reaction. In S. cerevisiae, ScPrp18 is non-essential for viability at growth temperatures <30°C (Vijayraghavan et al., 1989; Vijayraghavan and Abelson, 1990; Horowitz and Abelson, 1993b). The concerted action of ScSlu7 - ScPrp18 heteromeric complex is essential for proper 3’ss definition during the second catalytic reaction (Zhang and Schwer, 1997; James et al., 2002). These in vitro studies also hinted at a possible intron -specific requirement for ScPrp18 and ScSlu7 factors as they were dispensable for splicing of intron variants made in modified ACT1 intron containing transcripts (Brys and Schwer, 1996; Zhang and Schwer, 1997). A short spacing distance between branch point adenosine to 3’splice site rendered the substrate independent of Prp18 and Slu7 for the second step (Brys and Schwer, 1996; Zhang and Schwer, 1997). Extensive mutational analyses of budding yeast ScPrp18 identified two functional domains and suggested separate roles during splicing (Bacikova and Horowitz, 2002; James et al., 2002). Fission yeast with its genome harboring multiple introns and degenerate splice signals has recently emerged as a unique model to study relationships between splicing factors and their role in genomes with short introns. Previously, studies in our lab had initiated genetic and mutational analysis of S. pombe Prp18, the predicted homolog of budding yeast Prp18. Genetic analysis showed its essentiality, but a set of missense mutants based on studies of budding yeast ScPrp18 (Bacikova and Horowitz, 2002) gave either inactive null or entirely wild type phenotype for the fission yeast protein. In this study, we have extended our previous mutational analysis of fission yeast Prp18 by adopting biophysical and computational approaches to generate temperature-sensitive mutants. A missense mutant was used to understand the splicing functions and interactions of SpPrp18 and the findings are summarized below. Fission yeast SpPrp18 is an essential splicing factor with transcript-specific functions and links efficient splicing with cell cycle progression We initiated our analysis of SpPrp18 by adopting a biophysical approach to generate ts mutants. We used the PREDBUR algorithm to predict a set of buried residues, which when mutated could result in a temperature-sensitive phenotype that complements the null allele at permissive temperature. These predictions are based upon two biophysical properties of amino acids: 1) Hydrophobicity, which is calculated in a window of seven amino acids 2) Hydrophobic moment, which is calculated in a sliding window of nine amino acids in a given protein sequence. Several studies correlate these properties to protein stability and function (Varadarajan et al., 1996). One of the buried residue mutants V194R, in helix 1 of SpPrp18 conferred weak temperature- sensitivity and strong cold-sensitivity even when the protein was over expressed from a plasmid. Through semi-quantitative RT-PCR we showed splicing-defects for tfIId+ intron1 in these cells even when grown at permissive temperature. The primary phenotype was the accumulation of pre-mRNA. Further, we showed this splicing arrest is co-related with reduced levels of SpPrp18 protein, linking protein stability and splicing function. Next we examined the effects of this mutation on function by further reduction of protein levels. This was done by integrating the expression cassette nmt81:spprp18+/spprp18V194R at the leu1 chromosomal locus and by metabolic depletion of the integrated allele. Through RT-PCRs we demonstrated that depletion of wild type or missense protein has intron specific splicing defects. These findings showed its non-global and possibly substrate-specific splicing function. In the affected introns, precursor accumulation is the major phenotype, confirming prior data from our lab that hinted at its likely early splicing role. This contrasts with the second step splicing role of the human or budding yeast Prp18 proteins. Previous data from our lab showed loss of physical interaction between SpPrp18 and SpSlu7 by co-immunoprecipitation studies. This again differs from the strong and functionally important ScPrp18 and ScSlu7 interaction seen in budding yeast. We show the absence of charged residues in SpSlu7 interaction region formed by SpPrp18 helix1 and helix2 which can explain the altered associations for SpPrp18 in fission yeast. Importantly, as the V194R mutation in helix 1 shows splicing defects even at permissive temperature, the data indicate a critical role for helix 1 for splicing interactions, possibly one that bridges or stabilizes the proposed weak association of SpPrp18-SpSlu7 with a yet unknown splicing factor. We also investigated the effects of mutations in other helices; surprisingly we recovered only mutations with very subtle growth phenotypes and very mild splicing defects. Not surprisingly, stop codon at L239 residue predicted to form a truncated protein lacking helices 3, 4 and 5 conferred recessive but null phenotype implicating essential functions for other helices. Other amino acid substitutions at L239 position had near wild type phenotype at 30°C and 37°C. Helix 3 buried residue mutant I259A conferred strong cold-sensitivity when over expressed from plasmid, but semi quantitative analysis indicated no splicing defects for intron1 in the constitutively expressed transcript tfIId+. These findings indicate cold sensitivity either arises due to compromised splicing of yet unknown transcripts or that over-expressed protein has near wild type activity. We find mutations in the helix 5 buried residues L324 also conferred near WT phenotype. Earlier studies in the lab found that substitution of surface residues KR that are in helix 5 with alanine lead to null phenotypes (Piyush Khandelia and Usha Vijayraghavan unpublished data). We report stable expression of all of these mutant proteins; L239A, L239P, L239G, I259A, I259V, L324F, L324A as determined by our immunoblot analysis at 30°C and 37°C. The mild phenotypes of many buried residues can be attributed to orientation of their functional groups into a protein cavity between the helices. Lastly, our microscopic cellular and biochemical analysis of cellular phenotypes of spprp18 mutant provided a novel and direct role of this factor in G1-S transition of cell cycle. Our RT-PCR data suggest spprp18+ is required for efficient splicing of several intron containing transcripts involved in G1-S transition and subsequent activation of MBF complex (MluI cell cycle box-binding factor complex) during S-phase and shows a mechanistic link between cell cycle progression and splicing. A tool to study links between RNA interference, centromeric non-coding RNA transcription and heterochromatin formation S.pombe possesses fully functional RNA interference machinery with a single copy for essential RNAi genes ago1+, dcr1+ and rdp1+. Deletion of any of these genes causes loss of heterochromatinzation with abnormal cytokinesis, cell-cycle deregulation and mating defects (Volpe et al., 2002). In S.pombe, exogenous or endogenously generated dsRNA’s from transcription of centromeric repeats are processed by the RNaseIII enzyme dicer to form siRNA. These siRNA’s are loaded in Ago1 to form minimal RNA induced silencing complex (RISC) complex or specialized transcription machinery complex RNA induced transcriptional silencing (RITS) complex and target chromatin or complementary mRNAs for silencing. Thus as in other eukaryotes, fission yeast cells deploy RNAi mediated silencing machinery to regulate gene-expression and influence chromatin status. Several recent studies point to emerging new roles of RNAi and its association with other RNA processes (Woolcock et al., 2011; Bayane et al., 2008; Kallgren et al., 2014). Many recent reports suggest physical interactions of RISC or RITS and RNA dependent RNA polymerase complex (RDRC) with either some factors of the spliceosomal machinery, heterochromatin machinery (CLRC complex) and the exosome mediated RNA degradation machinery (Bayne et al., 2008 and Chinen et al., 2010 ; Hiriart et al., 2012; Buhler et al., 2008; Bayne et al., 2010 ). Thus we presume conditional alleles in spago1+ will facilitate future studies to probe the genetic network between these complexes as most analyses thus far rely on ago1∆ allele or have been based on proteomic pull down analyses of RISC or RITS complexes. In this study, we employed biophysical and modeling approaches described earlier to generate temperature sensitive mutants in spago1+ and spdcr1+. We tested several mutants for their ability to repress two reporter genes in a conditional manner. Our modeling studies on SpAgo1 PAZ domain indicated structural similarities with human Ago1 PAZ domain. We created site-directed missense mutants at predicted buried residues or in catalytic residues. We also analyzed the effects of random amino acid replacements in specific predicted buried or catalytic residues of SpAgoI. These ago1 mutants were screened as pools for their effects on silencing of GFP or of ura4+ reporter genes. These assays assessed post transcriptional gene silencing (PTGS) or transcriptional gene silencing (TGS) activity of these mutants. We obtained three temperature sensitive SpAgo1 mutants V324G, V324S and L215V while the V324E replacement was a null allele. Based upon our modeling, a likely explanation for the phenotype of these mutants is structural distortion or mis-orientation of the functional groups caused due to these mutations, which affect activity in a temperature dependent manner. This distortion in the PAZ domain may affect binding of siRNA and thereby lead to heterochromatin formation defects that we observed. Our data on the SpAgo1 V324 mutant shows conditional centromeric heterochromatin formation confirmed by semi quantitative RT-PCR for dh transcripts levels that shows temperature dependent increase in these transcripts. We find reduced H3K9Me2 levels at dh locus by chromatin immunoprecipitation (ChIP) assay, linking the association of siRNAs for establishment of heterochromatin at this loci. The data on PTGS of GFP transcripts show SpAgo1 V324G mutation has decreased slicing activity as semi-quantitative RT-PCR for GFP transcripts show increased levels at non permissive temperature. These studies point out the importance of siRNA binding to the PAZ domain and its effect on slicing activity of SpAgo1. The mutations in Y292 showed residue loss of centromeric heterochromatin formation phenotype. Thus, we ascribe critical siRNA binding and 3’ end recognition functions to this residue of SpAgo1. These studies point out functional and structural conservation across hAgo1 and SpAgo1. Adopting the aforementioned biophysical mutational approach, we generated mutants in spdcr1+ and screened for those with conditional activity. Our modeling studies on SpDcr1 helicase domain shows it adopts the conserved helicase domain structure seen for other DEAD Box helicases. Our data on mutational analysis of a conserved buried residue I143 in the walker motif B created inactive protein. The data confirm critical functions for dicer in generation of siRNAs and also in recognition of dsRNA ends. Mutants in buried residues L1130 and I1228 of RNase IIIb domain were inactive and the proximity of these residues to the catalytic core suggest that the critical structural alignment of catalytic residues is indispensable for carrying out dsRNA cleavage to generate siRNAs. We also attribute critical catalytic functions to SpDcr1 D1185 residue for generation of siRNA and heterochromatin formation as measured by our transcriptional gene silencing assay. Our studies employing biophysical and computational approaches to design temperature-sensitive mutants have been successfully applied to an essential splicing factor SpPrp18, which was refractory for ts mutants by other methods. Using a missense mutant, we showed its intron-specific splicing function for subsets of transcripts and deduced that its ubiquitous splicing role is arguable. We have uncovered a link between the splicing substrates of SpPrp18 and direct evidence of splicing based cell cycle regulation, thus providing a mechanistic link to the cell cycle arrest seen in some splicing factor mutants. The same methodology was applied to another important biological pathway, the RNAi machinery, where central factors SpAgoI and SpDcrI were examined We report the first instance of conditional gene silencing tool by designing Ago1 ts mutants which will be useful for future studies of the global interaction network between RNAi and other RNA processing events.

Les structures d'acto-myosine sont impliquées dans de nombreuses fonctions cellulaires. Comprendre leur organisation et leur comportement collectif est toujours difficile. Nous avons étudié l'anneau cytokinétique dans les cellules de mammifères et dans les levures de fission, en orientant les cellules dans les microcavités, ce qui permet de voir l'anneau dans un seul plan focal. Avec cette configuration, nous révélons de nouvelles structures et des dynamiques distinctes pour les deux systèmes cellulaires. Dans les cellules de mammifères, nous trouvons des motifs réguliers de la myosine et la formine. Les caractéristiques de ces motifs sont stables tout au long de sa fermeture et leur apparition coïncide avec la constriction. Nous proposons que ce phénomène est une propriété inhérente du réseau d'acto-myosine et que la formation de ces motifs entraîne une augmentation du stress. Ces hypothèses sont confirmées par notre modèle en champ moyen. Par contraste, l'anneau de levure de fission montre des inhomogénéités tournantes de l'actine, de la myosine, des protéines de la construction de la paroi (Bgs) et d'autres protéines. La dynamique des inhomogénéités de myosine est inchangée, si la croissance de la paroi est inhibée. Cependant, l'inhibition du mouvement des inhomogénéités conduit à l'arrêt de la fermeture. Nous proposons que la fermeture de l'anneau est entraînée par la rotation de l'actine et de la myosine qui tirent des protéines Bgs, lesquelles construisent ainsi le septum. Cette hypothèse est confirmée par nos calculs et par des simulations numériques. Nous suggérons que la transition entre les états de différents ordres et dynamiques pourrait être une façon de réguler in vivo les systèmes d'acto-myosine. / Actomyosin structures are involved in many cell functions. Understanding their organization and collective behavior is still challenging. We study the cytokinetic ring in mammalian cells and in fission yeasts, by orienting cells in microcavities. This allows seeing the ring in a single plane of focus. With this setup, we reveal new structures and distinct dynamics for both cellular systems. In mammalian cells we find a pattern of regular clusters of myosin and formin. The characteristics of this pattern are stable throughout closure and its formation coincides with the onset of constriction. We propose that its characteristic is an inherent property of the actomyosin network and that its formation leads to an increase in stress generation. These hypotheses are supported by our theoretical mean field model. In contrast, fission yeast rings show rotating inhomogeneities (speckles), i.e. rotations of actin, myosin, cell wall building proteins (Bgs) and other proteins. Myosin speckles dynamic is unchanged, if wall growth is inhibited. However, the inhibition of speckle motion leads to stalled closure. We propose that the ring closure is driven by the rotation of actin and myosin, which pull Bgs thereby building the septum. This model is supported by our calculations and by numerical simulations. We suggest that the transition between states of different orders and dynamics might be a way to regulate actomyosin systems in vivo.