Titre de l'écran-titre (visionné le 26 juin 2023) / FMRP (Fragile X mental retardation protein) est une protéine de liaison à l'ARN dont l'absence cause le développement du syndrome du X fragile avec retard mental (FXS). Ceci serait dû en partie à la perte de la fonction traductionnelle des formes cytoplasmiques de la protéine. Des isoformes nucléaires de FMRP (nFMRP) ont également été identifiées. Cependant, leurs fonctions demeurent très peu étudiées. Ces isoformes ont été localisées au sein de structure nucléaire de traitement de l'ARN, les corps de Cajal. De plus, nos investigations ont impliqué nFMRP dans la réponse cellulaire aux dommages à l'ADN identifiant nFMRP comme étant un antagoniste de la formation de structures d'instabilité génomique et empêchant l'accumulation des dommages à l'ADN associé. Toutefois, les mécanismes sous-jacents impliquant nFMRP et ses interacteurs demeurent inconnus. L'hypothèse est que nFMRP interagit avec des partenaires nucléaires impliqués dans le métabolisme de l'ARN et dans la signalisation du dommage à l'ADN afin de maintenir l'intégrité génomique cellulaire. Les objectifs sont d'identifier les partenaires de nFMRP chez l'humain et de caractériser leur interaction. Ce travail a permis de localiser nFMRP au sein de sites de réparation du dommage à l'ADN, les foyers de Fanconi et d'investiguer l'effet de la protéine sur les R-loops. Finalement, mes travaux ont identifié la protéine DDX5, une hélicase à ARN, réparant les dommages à l'ADN par la résolution de R-loops, comme étant le premier partenaire de nFMRP. Ce résultat permet de définir le rôle joué par nFMRP dans le maintien de la stabilité génomique. Des études de mutagenèses permettront de caractériser le mécanisme entre nFMRP et DDX5 et de futures analyses de protéomique à large spectre permettront d'établir le premier interactome de nFMRP. Ce travail permettra d'ouvrir des avenues de recherche dans la perspective de mieux comprendre les fonctions de nFMRP ainsi que la physiopathologie du FXS. / FMRP (Fragile X mental retardation protein) is an RNA-binding protein whose absence causes the development of Fragile X syndrome with mental retardation (FXS). This would be due in part to the loss of the translational function of the cytoplasmic forms of the protein. Nuclear isoforms of FMRP (nFMRP) have also been identified. However, their functions remain very little studied. These isoforms have been localized within the nuclear RNA processing structure, the Cajal bodies. Furthermore, our investigations implicated nFMRP in the DNA damage cellular response, identifying nFMRP as an antagonist of the formation of genomic instability structures and preventing the accumulation of associated DNA damage. However, the underlying mechanisms involving nFMRP and its interactors remain unknown. The hypothesis is that nFMRP interacts with nuclear partners involved in RNA metabolism and DNA damage signaling to maintain cellular genomic integrity. The objectives are to identify the partners of nFMRP in humans and to characterize their interaction. This work made it possible to locate nFMRP within DNA damage repair sites, the Fanconi foci, and to investigate the effect of the protein on R-loops. Finally, my work identified the protein DDX5, an RNA helicase, repairing DNA damage by resolving R-loops, as the first partner of nFMRP. This result makes it possible to define the role played by nFMRP in the maintenance of genomic stability. Mutagenesis studies will characterize the mechanism between nFMRP and DDX5 and future broad-spectrum proteomic analyzes will establish the first interactome of nFMRP. This work will open avenues of research to better understanding the functions of nFMRP as well as the pathophysiology of FXS.

This thesis examines the use and function of a specific group of Psalms, the so-called “Royal Psalms,” among the texts of the Qumran library. From the time of their integration into the worship practices of the Israelite people in the obscure past to the Second Temple period and beyond, these Psalms continued to be a source of inspiration to the Jewish people. Though there have been many studies that have analyzed their Sitz im Leben, use, interpretation, and application for many different periods, no study has attempted a thorough analysis of their use among the Qumran documents. Analyses of the use in the Qumran texts of certain individual Royal Psalms exist, but these do not attempt to cover the Royal Psalms as a corpus. The present thesis will analyze the appearance in the Qumran library of the eleven generally-accepted Royal Psalms: Pss 2, 18, 20, 21, 45, 72, 89, 101, 110, 132, and 144. This study explores whether or not these Psalms are to be found in the known Qumran Psalms scrolls, variations or differences as compared to the Masoretic Text, how they are were interpreted in exegetical and other texts, quotations of and allusions to them, and how themes from the Royal Psalms contribute to the structure and theology of non-canonical royal psalms found at Qumran. An understanding of the use of the biblical Royal Psalms in these texts is of value for our comprehension of what happened to the pre-exilic royal traditions as these hymns continued to be used in a post-monarchic society. This dissertation makes an original contribution toward these goals, establishing that there was an interest on the part of the authors of many of the Qumran texts in royal themes although they lived long after the monarchy had ended.

Com a construção da linhagem RlhE fusionada a um epitopo, fomos capazes de investigar o acúmulo da proteína após choque frio, em fase estacionária, durante o ciclo celular de Caulobacter crescentus, sob diferentes condições de estresses e, finalmente, após a adição de cloranfenicol, para o estudo a estabilidade protéica. Foi observado um aumento no acúmulo da proteína após choque frio. Além disso, quando em diferentes condições de estresses, RlhE obteve uma leve indução na presença de NaCl e Sacarose; mas permanaceu constante em fase estacionária e durante o ciclo celular. Finalmente, vimos que a estabilidade de RlhE varia de acordo com a temperatura, tendo um aumento da estabilidade a 10°C. As análises de expressão do gene rhlE foram realizadas por ensaios de atividade de betagalactosidase. Demonstramos que a presença da região 5UTR é importante para a indução, e que rlhE possui, pelo menos em parte, uma regulação pós-transcricional. Uma análise transcriptômica global da linhagem selvagem e mutante para rhlE, após o choque frio, foi realizada por RNAseq, o qual nos auxiliou na identificação de genes envolvidos em diversos processos biológicos. Finalmente, a co-imunoprecipitação e identificação dos RNAs por sequenciamento em larga escala revelou que RlhE interage com 51 mRNAs. / Here, we constructed a strain in which the RhIE protein was fusioned to an epitope that allowed the investigation of the protein profile after cold-shock, at stationary phase, during cell cycle of Caulobacter crescentus, under different environmental stresses, and, finally, after chloramphenicol addition to study protein stability. The results showed an increase in protein concentration after cold shock stress. When exposed to different stresses RhlE was slightly induced in the presence of NaCl and Sucrose; but its abundance remained constant at stationary phase and during C. crescentus cell cycle. Lastly, the protein stability varies depending on temperature - increasing at low temperature. Gene expression analysis was performed using beta-galactosidase assays. We showed that the presence of the 5UTR is important for the induction of rhlE, and that RhlE is posttranscriptionally regulated. A global transcriptome analysis was performed using RNAseq after cold shock stress of the wild type strain and null mutant for rhlE, and several genes involved in a wide range of biological process were identified. Finally, High-throughput sequencing of RNA isolated by crosslinking immunoprecipitation revealed interactions of RhlE with 51 mRNAs.

L’ARN hélicase à boite DEAD de la levure S.cerevisiae Ded1 est une protéine essentielle dont la fonction a été conservée au cours de l’évolution. Ses homologues fonctionnels sont impliqués dans le développement et le cycle cellulaire. Ded1 a longtemps été associée à l’étape de scanning de la région 5’UTR des ARNm au niveau de l’initiation de la traduction. Nous avons utilisé différentes approches comme les co-immunoprécipitations, des analyses de spectrométrie de masse, des tests de complémentation génétique, de séparation des complexes sur gradients de saccharose, des expériences de localisation in situ et d’enzymologie pour montrer que Ded1 interagissait physiquement avec des complexes cytoplasmique et nucléaire de liaison à la coiffe des ARNm. Nous avons également montré que Ded1 peut passer du noyau vers le cytoplasme par différentes voies d’export nucléaire. De façon intéressante, ses partenaires protéines sont capables de stimuler son activité ATPase. De plus, nous avons montré qu’il existait un lien génétique entre Ded1 et ses partenaires. Nous avons également montré que Ded1 colocalise partiellement avec ses partenaires dans des gradients de saccharose, suggérant que Ded1 pourrait être associée à certains mRNPs. Nos résultats encore préliminaires indiquent que Ded1 pourrait s’associer à d’autre ARNs coiffés. Ainsi, Ded1 pourrait remodeler les complexes associés à différentes étapes de la vie des ARN coiffés. / The budding yeast DEAD-box RNA helicase Ded1 is an essential yeast protein that is closely related to a subfamily of DEAD-box proteins that are involved in developmental and cell-cycle regulation. Ded1 is generally considered to be a translation-initiation factor that helps the 40S ribosome scan the mRNA from the 5' 7-methylguanosine cap to the AUG start codon. We have used IgG pulldown experiments, mass spectroscopy analyses, genetic experiments, saccharose gradients, in situ localizations, and enzymatic assays to show that Ded1 is a cap-associated factor that actively shuttles between the cytoplasm and the nucleus. We show that Ded1 physically interacts with various cap-associated factors and that its enzymatic activity is stimulated by these factors. By using various mutated proteins, we show that Ded1 is genetically linked to these factors. Ded1 comigrates with these factors on saccharose gradients, but the peak of Ded1 sediments slightly heavier than for the other factors, which suggests that Ded1 is predominately associated with a subset of the mRNPs. Finally, purification of the protein complexes associated with Ded1 and subsequent analysis by nanoLC-MS/MS indicates that Ded1 is associated with both nuclear and cytoplasmic mRNPs. Preliminary experiements showed that Ded1 can associate with other capped RNA. We conclude that Ded1 may function as a remodeling factor that is needed to form the different complexes associated with the different processing steps of the capped RNA.

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A concept of underworld runs through many cultures. These realms of spirits and the dead generally share several key characteristics, despite their varied and separate traditions. The commonalities among these different mythological places make it possible to generalize certain characteristics as chthonic for descriptive and analytical purposes. The underworld has appeared widely in video games throughout their history. I argue that the remarkable prevalence reflects a formal relationship between the underworld and video games; specific elements in mythic underworlds comprise a chthonic poetics that resonates with video game worlds and affordances. Video games uniquely support the spatial, thematic, and narrative elements that characterize underworlds and the philosophical questions they engage. Embodiment binds these elements together; they are unintelligible without this core perspective. The body sits at the axis of experience in mythic underworlds like the one described by Dante in the Divine Comedy and in game worlds like the one in World of Warcraft. It provides the medium through which we can experience these simulations as worlds rather than mere information structures. Other formal elements give context and direction to that embodied experience, and exploring how these interact with embodiment can expand our understanding of chthonic embodiment and the experience of space in virtual worlds. Three primary forces acting on the agency, subjectivity and control of the body structure that experience, which in turn reflects the ways of being that emerge from chthonic contexts. By incorporating these forces into their gameplay and narrative structure, games provide more direct access to the mythic power associated with the underworld than previous media forms.

Factual accounts of revenants – the risen dead – seized the medieval imagination in the early eleventh century, and were recorded by serious historians and ecclesiastics as true. They then began to appear in secular imaginative literature and art, growing progressively more elaborate and frightening throughout the Middle Ages whilst retaining many of the religious overtones expressed overtly in the ecclesiastic tales. By the early modern and modern period, the tales were removed from any overt religious context and were told as purely imaginative literature. The academic half of this thesis explores the influence on the tales of the Christian doctrine of resurrection and the cult of the body of Christ and of the saints, then traces the migration of those tales into imaginative literature from the Middle Ages to the present. It identifies key motifs from the medieval chronicles and imaginative literature that continue to appear in modern stories, and explores the extent to which Christian eschatology altered perceptions of the dead and why, in an increasingly secular context, fascination with such tales continued into modern literature, what part fear of death played throughout this period, and how that fear was expressed, first in an ecclesiastical context, then in imaginative literature through horror stories. The creative half of my thesis is a literary fiction novel updating a medieval revenant tale, the Legend of the Three Living and the Three Dead, to twenty-first century New England.

The inverter dead-time is integral to the safety of a voltage source inverter (VSI). Dead-time is introduced between the complementary gating signals of the top and bottom switches in each VSI leg to prevent shoot-through fault. This thesis reports and investigates dead-time induced sub-harmonic oscillations in open-loop induction motor drives of different power levels, under light-load conditions. The thesis develops mathematical models that help understand and predict the oscillatory behaviour of such motor drives due to dead-time act. Models are also developed to study the impact of under-compensation and over-compensation of dead-time act on stability. The various models are validated through extensive simulations and experimental results. The thesis also proposes and validates active damping schemes for mitigation of such sub-harmonic oscillations. The thesis reports high-amplitude sub-harmonic oscillations in the stator current, torque and speed of a 100-kW open-loop induction motor drive in the laboratory, operating under no-load. Experimental studies, carried out on 22-kW, 11-kW, 7.5-kW and 3.7-kW open-loop induction motor drives, establish the prevalence of dead-time induced sub-harmonic oscillations in open-loop motor drives of different power levels. An experimental procedure is established for systematic study of this phenomenon in industrial drives. This procedure yields the operating region, if any, where the motor drive is oscillatory. As a first step towards understanding the oscillatory behaviour of the motor drive, a mathematical model of the VSI is derived in a synchronously revolving reference frame (SRF), incorporating the of dead-time on the inverter output voltage. This leads to a modified dynamic model of the inverter-fed induction motor in the SRF, inclusive of the dead-time act. While the rotor dynamic equations are already non-linear, dead-time is found to introduce nonlinearities in the stator dynamic equations as well. The nonlinearities in the modified dynamic model make even the steady solution non-trivial. Under steady conditions, the dead-time can be modelled as the drop across an equivalent resistance (Req0) in the stator circuit. A precise method to evaluate the equivalent resistance Req0 and a simple method to arrive at the steady solution are proposed and validated. For the purpose of stability analysis, a small-signal model of the drive is then derived by linearizing the non-linear dynamic equations of the motor drive, about a steady-state operating point. The proposed small-signal model shows that dead-time contributes to different values of equivalent resistances along the q-axis and d-axis and also to equivalent cross-coupling reactance’s that appear in series with the stator windings. Stability analysis performed using the proposed model brings out the region of oscillatory behaviour (or region of small-signal instability) of the 100-kW motor drive on the voltage versus frequency (V- f) plane, considering no-load. The oscillatory region predicted by the small-signal analysis is in good agreement with simulations and practical observations for the 100-kW motor drive. The small-signal analysis is also able to predict the region of oscillatory behaviour of an 11-kW motor drive, which is con consumed by simulations and experiments. The analysis also predicts the frequencies of sub-harmonic oscillations at different operating points quite well for both the drives. Having the validity of the small-signal analysis at different power levels, this analytical procedure is used to predict the regions of oscillatory behaviour of 2-pole, 4-pole, 6-pole and 8-pole induction motors rated 55 kW and 110 kW. The impact of dead-time on inverter output voltage has been studied widely in literature. This thesis studies the influence of dead-time on the inverter input current as well. Based on this study, the dynamic model of the inverter fed induction motor is extended to include the dc-link dynamics as well. Simulation results based on this extended model tally well with the experimentally measured dc-link voltage and stator current waveforms in the 100-kW drive. Dead-time compensation may be employed to mitigate the dead-time and oscillatory behaviour of the drive. However, accurate dead-time compensation is challenging to achieve due to various factors such as delays in gate drivers, device switching characteristics, etc. Effects of under-compensation and over-compensation of dead time are investigated in this thesis. Under-compensation is shown to result in the same kind of oscillatory behaviour as observed with dead-time, but the fundamental frequency range over which such oscillations occur is reduced. On the other hand, over-compensation of dead-time effect is shown to result in a different kind of oscillatory behaviour. These two types of oscillatory behaviour due to under- and over-compensation, respectively, are distinguished and demonstrated by analyses, simulations and experiments on the 100-kW drive. To mitigate the oscillatory behaviour of the drive, an active damping scheme is proposed. This scheme emulates the effect of an external inductor in series with the stator winding. A small-signal model is proposed for an induction motor drive with the proposed active damping scheme. Simulations and experiments on the 100-kW drive demonstrate effective mitigation of light-load instability with this active damping scheme. In the above inductance emulation scheme, the emulated inductance is seen by the sub-harmonic components, fundamental component as well as low-order harmonic components of the motor current. Since the emulated inductance is also seen by the fundamental component, there is a fundamental voltage drop across the emulated inductance, leading to reduced co-operation of the induction motor. Hence, an improved active damping scheme is proposed wherein the emulated inductance is seen only by the sub-harmonic and low-order harmonic components. This is achieved through appropriate altering in the synchronously revolving domain. The proposed improved active damping scheme is shown to mitigate the sub-harmonic oscillation effectively without any reduction in flux.