Chlamydia trachomatis is a Gram-negative, obligate intracellular pathogen that is the causative agent of sexually transmitted infections and the ocular disease trachoma. Chlamydia trachomatis undergoes a biphasic developmental cycle differentiating between the infectious elementary body (EB) and the replicative reticulate body (RB). Under certain stress conditions, C. trachomatis can stall its developmental cycle and enter an aberrant state termed persistence. While in a persistent state, C. trachomatis is refractory toward antibiotics, can evade the host immune response, and becomes undetectable using standard clinical detection methods. Environmental and other pathogenic microbes are known to utilize partner switching mechanisms (PSM) to regulate sigma factors used to initiate a stress response. For this reason, this study focuses on the chlamydial PSM, its role in regulating the availability of the housekeeping sigma factor σ66, and its role in the developmental cycle and stress response of C. trachomatis. The chlamydial PSM is composed of five known proteins: the anti-sigma factor RsbW, two anti-anti-sigma factors RsbV1 and RsbV2, a regulatory phosphatase RsbU, and a second phosphatase-like protein CTL0852. In order to test the role of the PSM in the chlamydial stress response, a panel of C. trachomatis rsbV1 mutants were generated, persistence inducing iron starvation and tryptophan starvation cell culture conditions were optimized, and growth of the rsbV1 mutants under iron starvation conditions were assayed. No significant differences were seen between rsbV1 mutants under iron starvation nor recovery conditions as determined by progeny production and inclusion size analysis. Furthermore, this study generated PSM protein producing Escherichia coli strains for in vitro protein work and performed operon mapping of the PSM genes of C. trachomatis to help aid in future studies of the chlamydial PSM by facilitating the development of new chlamydial PSM mutants. This study gives phylogenetic support to the classification of ctl0852 as a chlamydial PSM gene by comparing relative mutations rates of PSM genes across chlamydial species.

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La photosynthèse oxygénique peut être le lieu de formation des Formes Réactives de l’Oxygéne (FROs). Les FROs altèrent toutes les macromolécules de la cellule, générant ainsi un stress oxydant. Toute perturbation du métabolisme cellulaire peut conduire à ce type de stress. Les cyanobactérie sétant les premiers organismes à avoir émis de l’oxygène sur terre, elles ont du développer très tôt au cours de l’évolution des mécanismes de perception et de défense pour lutter contre ce stress. Nous nous sommes intéressés à l’étude des mécanismes qui permettent à la cyanobactérie filamentuse et diazotrophe Anabaena PCC 7120 de s’adapter à diverses conditions de stress et de carence : stress oxydant, carence en fer et en azote combiné. En réponse à une carence en azote combiné, elle différencie en 24 h des hétérocystes : cellules spécialisées dans la fixation de l’azote atmosphérique. Nous avons étudié la réponse transcriptomique globale de cette bactérie à la fois au stress oxydant et à la carence en fer et nous avons établit la connection existant entre ces deux stress. Nous avons pu identifier le régulateur transcriptionnel pleiotrope impliqué dans la perception et la signalisation du stress peroxyde et nous en avons élucidé le mécanisme d’action. Nous avons également étudié une Ser/Thr kinase qui joue un rôle important à la fois dans la réponse au stress oxydant et à la carence en azote combiné. Notre étude a montré que cette kinase pourrait être le lien moléculaire entre ces deux conditions, puisque une cible potentielle de cette kinase semble être la protéine HetR qui est le régulateur clé du processus de différenciation cellulaire. / Oxygenic photosynthesis may generate Reactive forms of Oxygéne (ROS). These reactive oxygen species can damage all the macromolecules of the cell, inducing oxidative stress. Any disruption of cellular metabolism can lead to oxidant damage. Cyanobacteria were the first organisms producing oxygen on Earth, they therefore had to develop very early during evolution the mechanisms of perception and defence to cope with this tstress. We are interested in studying the mechanisms that allow the diazotrophic filamentous cyanobacterium Anabaena PCC 7120 to adapt to various conditions of stress and stravations: oxidative stress, iron and combined nitrogen starvations. Anabaena PCC 7120 is a simple model for the study of cell differentiation. In response to combined nitrogen stravation it can differentiate heterocysts, cell specialized in molecular nitrogen fixation. We studied the global transcriptomic response of this bacterium to both oxidative stress and iron deficiency and we establish the crosstalk between these two stresses. We were able to identify the global transcriptional regulator involved in the perception and in the signaling of a peroxide stress. Its mechanism of action was elucidated. We also studied a Ser / Thr kinase that plays an important role both in the response to oxidative stress and combined nitrogen stravation. Our study showed that this kinase may be the molecular link between these two conditions, as a potential target of this kinase appears to be the HetR protein which is the key regulator of cellular differentiation process.

Fatores sigma de carência de ferro, representados por FecI de E. coli, direcionam a transcrição de genes de transporte de sideróforos (quelantes de ferro), e são geralmente regulados por fatores antissigma (FecR), que liberam o fator sigma após ligação do sideróforo no receptor de membrana externa (FecA). Caulobacter crescentus possui quatro genes para fatores sigma desta família. Ensaios de expressão gênica e crescimento indicaram que estes genes não respondem à disponibilidade de ferro. Em microarranjos de cDNA, apenas o gene fecA2 foi induzido em DfecR2 comparado à linhagem parental, sugerindo que este é o único gene alvo do fator sigma FecI2. Já DfecR4 mostrou indução em mais de 50 genes, alguns envolvidos na utilização de fontes alternativas de carbono. Ensaios fenotípicos com DfecI4 sugeriram que este gene é importante para o crescimento em g-ciclodextrina ou ácido caproico. Os resultados sugerem que o fator sigma FecI2 é bem específico, enquanto FecI4 parece regular uma resposta geral relacionada a compostos carbônicos, e não à homeostase de ferro. / Iron starvation sigma factors, whose prototype is E. coli FecI, direct transcription of genes involved in siderophore (iron chelators) transport, being usually regulated by anti-sigma factors (FecR), which release the sigma factor after siderophore binding to the outer membrane receptor (FecA). Caulobacter crescentus possesses four genes encoding FecI-like sigma factors. Gene expression and growth assays indicated that these genes do not respond to iron availability. In cDNA microarrays, only the fecA2 gene was induced in DfecR2 relative to the wild-type strain, suggesting that this is the only target gene of the FecI2 sigma factor. However, in DfecR4 there was induction of over 50 genes, some of them involved in utilization of alternative carbon sources. Phenotypic microarrays with the DfecI4 strain showed that this gene is important for growth in g-cyclodextrin or caproic acid. The results suggest that the FecI2 sigma factor is very specific, whereas FecI4 seems to regulate a more general response, related to carbon compounds rather than iron homeostasis.

Fatores sigma de carência de ferro, representados por FecI de E. coli, direcionam a transcrição de genes de transporte de sideróforos (quelantes de ferro), e são geralmente regulados por fatores antissigma (FecR), que liberam o fator sigma após ligação do sideróforo no receptor de membrana externa (FecA). Caulobacter crescentus possui quatro genes para fatores sigma desta família. Ensaios de expressão gênica e crescimento indicaram que estes genes não respondem à disponibilidade de ferro. Em microarranjos de cDNA, apenas o gene fecA2 foi induzido em DfecR2 comparado à linhagem parental, sugerindo que este é o único gene alvo do fator sigma FecI2. Já DfecR4 mostrou indução em mais de 50 genes, alguns envolvidos na utilização de fontes alternativas de carbono. Ensaios fenotípicos com DfecI4 sugeriram que este gene é importante para o crescimento em g-ciclodextrina ou ácido caproico. Os resultados sugerem que o fator sigma FecI2 é bem específico, enquanto FecI4 parece regular uma resposta geral relacionada a compostos carbônicos, e não à homeostase de ferro. / Iron starvation sigma factors, whose prototype is E. coli FecI, direct transcription of genes involved in siderophore (iron chelators) transport, being usually regulated by anti-sigma factors (FecR), which release the sigma factor after siderophore binding to the outer membrane receptor (FecA). Caulobacter crescentus possesses four genes encoding FecI-like sigma factors. Gene expression and growth assays indicated that these genes do not respond to iron availability. In cDNA microarrays, only the fecA2 gene was induced in DfecR2 relative to the wild-type strain, suggesting that this is the only target gene of the FecI2 sigma factor. However, in DfecR4 there was induction of over 50 genes, some of them involved in utilization of alternative carbon sources. Phenotypic microarrays with the DfecI4 strain showed that this gene is important for growth in g-cyclodextrin or caproic acid. The results suggest that the FecI2 sigma factor is very specific, whereas FecI4 seems to regulate a more general response, related to carbon compounds rather than iron homeostasis.

Les gibbérellines (GA) sont des phytohormones qui régulent divers aspects du développement en réponse aux signaux endogènes et exogènes à la plante. Ainsi face à un stress, les niveaux de GA sont finement contrôlés, permettant une croissance adaptée aux contraintes environnementales. Au niveau moléculaire, les GA stimulent la croissance de la plante en s’opposant aux protéines DELLAs (DELLAs), facteurs nucléaires qui inhibent la croissance. Les DELLAs présentent plusieurs caractéristiques fonctionnelles notables, une activité transactivatrice et la capacité d’interagir avec d’autres protéines régulatrices, comme les répresseurs de la signalisation Jasmonate (JA), JA ZIM-domain (JAZ). Le génome d’Arabidopsis thaliana code pour cinq DELLAs possédant des fonctions redondantes et spécifiques. Le but de mon travail de thèse a été la caractérisation de la fonction biologique d’une DELLA, RGA-LIKE3 (RGL3). J’ai pu montrer que RGL3 modifie la défense de la plante face à des stresses biotiques. / The phytohormones gibberellins (GA) regulate major aspects of plant growth in response to endogenous and environmental signals. Upon the perception of stress, the levels of bioactive GA are adjusted, hence allowing a flexible growth response to environmental variability. At a molecular level, GA promote growth by stimulating the degradation of the growth repressing DELLA proteins. DELLAs are versatile nuclear proteins with several remarkable features, such as transactivation activity and protein–protein interaction capacities. Thus DELLAs interact with a series of highly divergent proteins, including different transcription factor families, but also the Jasmonate (JA) ZIM-domain (JAZ) proteins, repressors of JA signaling. The aim of this thesis work consisted in the characterization of the biological function of the DELLA RGA-LIKE3. I could show that RGL3 modulates plant defense responses against biotic stresses.