Corticosteroid ligands activate the glucocorticoid receptor (GR). GR plays a role in glucose homeostasis, adipogenesis, inflammation, and mood and cognitive functions. Understanding the interplay of diverse forms of receptor regulation (including post-translational modification, cofactor interactions, ligand binding, and receptor localization) and their effects is important for understanding and developing more effective treatment for a variety of conditions. Prior to ligand binding, the naïve GR is primarily cytoplasmic, residing in a chaperone complex containing heat-shock proteins and immunophilins. Upon ligand-binding, alterations to the complex allow the receptor to dimerize and import into the nucleus. Nuclear GR interacts with transcriptional regulatory sequences and recruits cofactors to regulate specific gene expression. Upon hormone withdrawal, the original chaperone complex is reassembled and the receptor is exported to the cytoplasm. Interestingly, while the import of GR into the nucleus occurs very rapidly (t ½ = 5 min), the re-export is significantly slower (t ½ = 12-24h). Previous work by our lab and others has indicated the existence of a nuclear retention signal (NRS) within the GR. The NRS sequence of the GR, its interaction partners, and the role it might play in the activity of the receptor have not yet been fully defined. Work in the Hache lab indicates that mutation of the GR nuclear localization signal 1 (NL1) increases the export rate of nuclear GR to the cytoplasm, as well as compromising receptor import, suggesting that the NL1 overlapped an NRS sequence. In this work, I made a series of GR mutants, based on sequence from the SV40 large T antigen NLS, which lacks nuclear retention activity. Using these mutants, I found that GR nuclear retention is influenced by both specific residues within the hinge region and the location of the sequence within the receptor, as reintroduction of the NLS sequence at the N-terminus of the receptor retention mutant failed to reconstitute the retention activity. Agonist liganded and hormone-withdrawn receptor mutants showed a similar decrease in retention. By contrast, antagonist-withdrawn GR mutants were retained in the nucleus, possibly due to altered receptor configuration and interactions. Assays of GR-responsive promoter activation by receptor retention mutants showed that while no difference in the ability of retention mutants to activate transcription was seen at a simple promoter, activation of a complex promoter was compromised. This impaired transactivation for the SV506-523 mutant correlated with decreased histone H4 acetylation and PolII recruitment, while GR DNA-binding at the target promoter appeared to be unaffected. These results suggested that promoter-specific cofactor interactions might be implicated in GR nuclear retention. Loss of GR hinge interaction with Oct cofactors produced an incomplete loss of retention, suggesting overlapping signals, but not supporting Oct as a primary factor in GR retention. The overlap between important residues in GR nuclear retention and localization signals and the lack of retention shown by the SV40 NLS suggested that retention might be intrinsic to the sequence of particular NLS. Preliminary results suggest that the KT511-512 residues of GR may be of general importance in protein nuclear retention, while the role of proline is likely more variable. My research has focused on increasing our understanding of glucocorticoid receptor nuclear retention and its possible implications. I have determined that the KT511-512 residues of GR play an important role in its retention, and possibly also figure in nuclear retention of other proteins. These residues are involved in interactions which affect promoter-specific histone acetylation and transcriptional activation in GR, suggesting a reason for the existence of nuclear retention.

Les nucléomodulines sont des protéines produites par des bactéries parasites intracellulaires et qui sont importées dans le noyau des cellules infectées pour y moduler l'expression génique et contribuer ainsi à la virulence de la bactéries. L'identification de nucléomodulines chez plusieurs espèces de bactéries pathogènes a fait émerger ce concept comme une stratégie supplémentaire utilisée par les parasites intracellulaires pour contourner les moyens de défense de l'hôte. Le but de cette thèse était d'identifier et d'analyser d'éventuelles nucléomodulines produites par l'agent étiologique de la tuberculose, la bactérie Mycobacterium tuberculosis (Mtb). Nous avons tout d'abord analysé le génome de Mtb, à la recherche de gènes dont les produits portent des séquences analogues aux séquences de localisation nucléaire des eucaryotes (NLS). Nous avons pu identifier deux gènes de Mtb, Rv0229c et Rv3876, qui codent des protéines sécrétées dans le milieu de culture et qui se localisent dans le noyau lorsqu'elles sont exprimées dans des cellules épithéliales ou dans des macrophages murins ou humains. Les NLS de ces deux protéines ont été identifiées et leur modification abolit la localisation nucléaire dans les cellules eucaryotes. Le gène Rv0229c est trouvé spécifiquement dans le génome des espèces pathogènes du complexe Mtb. Ce gène semble avoir été acquis récemment par l'ancêtre de Mtb, via un transfert génétique horizontal. Rv3876 est plus généralement distribué chez les mycobactéries, et appartient à une région génomique codant un système de sécrétion type VII, ESX1, essentiel pour la virulence de Mtb. Les travaux en cours visent à analyser la dynamique de ces deux protéines au cours de l'infection de macrophages ou de modèles animaux, et leur rôle en tant que modulateurs de l'expression génique des cellules infectées et dans la virulence de Mtb. / The nuclear targeting of bacterial proteins that modify host cell gene expression, the so-called nucleomodulins, has emerged as a novel mechanism contributing to virulence of several intracellular pathogens. The goal of this study was to identify nucleomodulins produced by Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB), and to investigate their role upon infection of the host. We first performed a screening of Mtb genome in search of genes encoding proteins with putative eukaryotic-like nuclear localization signals (NLS). We identified two genes of Mtb, Rv0229c and Rv3876, encoding proteins that are secreted in the medium by Mtb and are localized into the nucleus when expressed in epithelial cells or in human or murine macrophages. The NLSs of these two proteins were identified and found to be essential for their nuclear localization. The gene Rv0229c, a putative RNase, is present only in pathogen species of the Mtb complex and seems to have been recently acquired by horizontal gene transfer (HGT). Rv3876 appears more widely distributed in mycobacteria, and belongs to a chromosomal region encoding proteins of the type VII secretion system ESX1, essential for virulence. Ongoing studies are currently investigating the dynamics of these proteins upon infection of host cells, and their putative role in the modulation of host cell gene expression and Mtb virulence.

Corticosteroid ligands activate the glucocorticoid receptor (GR). GR plays a role in glucose homeostasis, adipogenesis, inflammation, and mood and cognitive functions. Understanding the interplay of diverse forms of receptor regulation (including post-translational modification, cofactor interactions, ligand binding, and receptor localization) and their effects is important for understanding and developing more effective treatment for a variety of conditions. Prior to ligand binding, the naïve GR is primarily cytoplasmic, residing in a chaperone complex containing heat-shock proteins and immunophilins. Upon ligand-binding, alterations to the complex allow the receptor to dimerize and import into the nucleus. Nuclear GR interacts with transcriptional regulatory sequences and recruits cofactors to regulate specific gene expression. Upon hormone withdrawal, the original chaperone complex is reassembled and the receptor is exported to the cytoplasm. Interestingly, while the import of GR into the nucleus occurs very rapidly (t ½ = 5 min), the re-export is significantly slower (t ½ = 12-24h). Previous work by our lab and others has indicated the existence of a nuclear retention signal (NRS) within the GR. The NRS sequence of the GR, its interaction partners, and the role it might play in the activity of the receptor have not yet been fully defined. Work in the Hache lab indicates that mutation of the GR nuclear localization signal 1 (NL1) increases the export rate of nuclear GR to the cytoplasm, as well as compromising receptor import, suggesting that the NL1 overlapped an NRS sequence. In this work, I made a series of GR mutants, based on sequence from the SV40 large T antigen NLS, which lacks nuclear retention activity. Using these mutants, I found that GR nuclear retention is influenced by both specific residues within the hinge region and the location of the sequence within the receptor, as reintroduction of the NLS sequence at the N-terminus of the receptor retention mutant failed to reconstitute the retention activity. Agonist liganded and hormone-withdrawn receptor mutants showed a similar decrease in retention. By contrast, antagonist-withdrawn GR mutants were retained in the nucleus, possibly due to altered receptor configuration and interactions. Assays of GR-responsive promoter activation by receptor retention mutants showed that while no difference in the ability of retention mutants to activate transcription was seen at a simple promoter, activation of a complex promoter was compromised. This impaired transactivation for the SV506-523 mutant correlated with decreased histone H4 acetylation and PolII recruitment, while GR DNA-binding at the target promoter appeared to be unaffected. These results suggested that promoter-specific cofactor interactions might be implicated in GR nuclear retention. Loss of GR hinge interaction with Oct cofactors produced an incomplete loss of retention, suggesting overlapping signals, but not supporting Oct as a primary factor in GR retention. The overlap between important residues in GR nuclear retention and localization signals and the lack of retention shown by the SV40 NLS suggested that retention might be intrinsic to the sequence of particular NLS. Preliminary results suggest that the KT511-512 residues of GR may be of general importance in protein nuclear retention, while the role of proline is likely more variable. My research has focused on increasing our understanding of glucocorticoid receptor nuclear retention and its possible implications. I have determined that the KT511-512 residues of GR play an important role in its retention, and possibly also figure in nuclear retention of other proteins. These residues are involved in interactions which affect promoter-specific histone acetylation and transcriptional activation in GR, suggesting a reason for the existence of nuclear retention.

Le travail de cette thèse s’est articulé autour de l’étude de CRMP5 au cours du développement du système nerveux central. Nous avons mis en évidence une interaction directe entre CRMP5 et la tubuline, conduisant à une inhibition de la pousse neuritique dans différentes lignées cellulaires, ainsi qu’à une inhibition d’élongation uniquement au niveau des dendrites et non de l’axone, dans des cultures primaires de neurones de l’hippocampe. De plus, nous avons montré que CRMP5 pouvait annuler l’action de CRMP2, connue pour promouvoir la pousse neuritique, de façon dominante mais dépendante de la présence sur CRMP5 du site de fixation à la tubuline. Contrairement à CRMP2, l’expression de CRMP5 étant transitoire pendant la différentiation neuronale, elle permettrait de restreindre de façon spatio-temporelle l’effet de CRMP2 sur la pousse neuritique, régulant ainsi la polarité neuronale. D’autre part, nous avons également rapporté la présence de CRMP5 au niveau mitochondrial où elle pourrait jouer un rôle dans le processus d’autophagie des mitochondries. Enfin, nous nous sommes intéressés à l’étude de la CRMP5 exprimée en conditions pathologiques, et nous avons observé une nouvelle localisation nucléaire de la protéine dans certaines cellules cancéreuses. Etant localisée dans plusieurs compartiments subcellulaires et impliquée dans différents mécanismes moléculaires, l’ensemble de ce travail décrit donc la protéine CRMP5 comme une protéine « multi-fonctionnelle ». / The purpose of this work is to focus on the study of CRMP5 during development of the central nervous system. We have demonstrated a direct interaction between CRMP5 and tubulin, leading to inhibition of neurite outgrowth in different cell lines, and inhibition of growth only at dendritic but not axonal level, in hippocampal neurons. Furthermore, we showed that CRMP5 could counteract the previously described CRMP2 effect on neurite outgrowth. The CRMP5 acted as a dominant signal to counteract CRMP2 outgrowth promotion and this function is also dependent on the tubulin-binding capacity of CRMP5.Unlike CRMP2, the CRMP5 expression being transient during neuronal differentiation, it would imply in the spatiotemporal regulation of the CRMP2 effect on neurite outgrowth, thereby regulating neuronal polarity. In another part, we also reported the presence of CRMP5 at mitochondrial level in vivo where it could play a role in mitochondrial autophagic process.Finally, we were interested in the study of CRMP5 expressed in pathological conditions, and we discovered a new nuclear localization of the protein in some cancer cells. Being localizedin several subcellular compartments and involved in different molecular mechanisms, this work describes CRMP5 as a "multi-functional" protein.

No description available.

Project Xanadu is a complex hypertext system, a predecessor of the World Wide Web, which has not been implemented in its full potential. The goal of the thesis is to describe features of Xanadu and to critically evaluate implementation options of these features in the current Web with benefits and limitations of this approach. The thesis introduces a historical context of hypertext systems and analyzes Project Xanadu from two viewpoints: as a set of abstract designs and as a series of software prototypes. The thesis defines main features and goals of Xanadu and recontextualizes them for the current Web using new web standards and technologies. Based on the proposed solutions a new design is conceived which implements features of Xanadu in the context of the Web.

Thèse numérisée par la Division de la gestion de documents et des archives de l'Université de Montréal.

Ce travail cherche a comprendre comment l'ajout d'une géométrie non euclidienne dans un problème de Schrödinger non linéaire influe sur l'existence et l'unicité des solutions explosives de masse critique. On s'inspire pour beaucoup des travaux de Merle et Raphaël sur la méthode de modulation des paramètres d'invariance géométrique pour une EDP qui possède de bonnes lois de conservations. On s'appuie ici plus particulièrement sur un article de Raphaël et Szeftel qui prouve l'existence et l'unicité d'une solution de masse critique en dimension 2 pour l'équation de Schrödinger non linéaire avec potentiel d'inhomogénéité devant la non-linéarité, et qui explose par ailleurs au maximum de l'inhomogénéité. Dans un premier temps, il s'agit de reprendre la méthode dans son ensemble afin de l'adapter à des cas où le Laplacien n'est plus plat, et est remplacé par un opérateur de type Laplace-Beltrami ou Laplacien généralisé. Ayant mis en avant le rôle de la courbure au point d'explosion, en termes de conditions sur les dérivées de termes métriques, on reprend dans un deuxième temps l'étude dans le cas plus général d'une variété riemannienne. Grâce à un ansatz sur la solution qui intègre maintenant la transformation induite par la métrique, on est capable d'énoncer un résultat d'existence et d'unicité en termes de conditions géométriques sur la variété elle même. Par soucis de simplicité, on se limite néanmoins au rôle local de la métrique, en la supposant globalement définie dans une certaine carte, et asymptotiquement équivalente a la métrique euclidienne. / The present work aims at investigating the effects of a non-euclidean geometry on existence and uniqueness results for critical blow up NLS solutions. We will use many ideas from the works of Merle and Raphaël, particularly ideas from modulation theory which describes a solution in terms of geometric invariants parameters. We will rely more specically on a paper from Raphaël and Szeftel for existence and uniqueness of a critical mass blow up solution in dimension two tothe nonlinear Schrödinger equation with inhomogeneous potential acting on the nonlinearity, and which blows up where the inhomogeneity reaches its maximum. At first, we consider a generalized Laplacian operator and deploy the classical ansatz method to point out difficulties inherited from the non-flat metric terms, and in particular the key role played by the curvature at the blow-up point. In a second part, we reproduce the method when modifying the geometrical ansatz on which the parametrix is constructed, and investigate more precisely what is needed for existence and then uniqueness when dealing with a Laplace-Beltrami operator associated to a riemannian manifold. For simplicity, we shall only consider the role of g locally around the blow up point we are constructing, by assuming g is globally defined in some map, and asymptotically equals the usual euclidean metric.

Thèse numérisée par la Division de la gestion de documents et des archives de l'Université de Montréal

Cette thèse est consacrée à l’étude des propriétés dynamiques des solutions de type soliton d'équations aux dérivées partielles (EDP) dispersives non linéaires. `A travers des exemples-type de telles équations, l'équation de Schrödinger non-linéaire (NLS), l'équation de Korteweg-de Vries généralisée (gKdV) et le système de Schrödinger, on traite du comportement des solutions convergeant en temps grand vers des sommes de solitons (multi-solitons). Dans un premier temps, nous montrons que dans une configuration symétrique, avec des interactions fortes, le comportement de séparation des solitons logarithmique en temps est universel à la fois dans le cas sous-critique et sur-critique pour (NLS). Ensuite, en adaptant les techniques précédentes à l'équation (gKdV), nous prouvons un résultat similaire de l'existence de multi-solitons avec distance relative logarithmique; pour (gKdV), les solitons sont répulsifs dans le cas sous-critique et attractifs dans le cas sur-critique. Finalement, nous identifions un nouveau régime de distance logarithmique où les solitons sont non-symétriques pour le système de Schrödinger non-intégrable; une telle solution n'existe pas dans le cas intégrable pour le système et pour (NLS). / This thesis deals with long time dynamics of soliton solutions for nonlinear dispersive partial differential equation (PDE). Through typical examples of such equations, the nonlinear Schrödinger equation (NLS), the generalized Korteweg-de Vries equation (gKdV) and the coupled system of Schrödinger, we study the behavior of solutions, when time goes to infinity, towards sums of solitons (multi-solitons). First, we show that in the symmetric setting, with strong interactions, the behavior of logarithmic separation in time between solitons is universal in both subcritical and supercritical case. Next, adapting previous techniques to (gKdV) equation, we prove a similar result of existence of multi-solitons with logarithmic relative distance; for (gKdV), the solitons are repulsive in the subcritical case and attractive in the supercritical case. Finally, we identify a new logarithmic regime where the solitons are non-symmetric for the non-integrable coupled system of Schrödinger; such solution does not exist in the integrable case for the system and for (NLS).