Dynamique des facteurs pré-ribosomiques au cours de la biogenèse de la grande sous-unité ribosomique chez S. cerevisiae

This work focuses on the dynamics of assembly, dissociation and recycling of proteins involved in the biogenesis of the large ribosomal subunit in Saccharomyces cerevisiae. It sheds some light on two control points in this metabolic pathway, localised in the nucleus and the cytoplasm respectively.We have shown that the nuclear protein Nsa2, which is very conserved throughout the eukaryotic kingdom, is required for the correct maturation of the 27SB ribosomal RNA precursor. Nsa2 is an unstable factor, regulated in correlation with the activity of ribosome biogenesis; it thus constitutes a good candidate for the integration of various signals resulting in the regulation of this metabolic pathway. Besides, using the SILAC technique, we could define groups of early or late acting factors relative to the Nsa2 action time.In the cytoplasm, we identified a protein network, which marks the end of ribosome biogenesis and triggers the entry of new ribosomal subunits into translation. The cytoplasmic protein Rei1 and the karyopherin Kap121 are both required for the recycling from the cytoplasm to the nucleus of a dimer of shuttling factors, Arx1-Alb1. This recycling enables the dissociation of the anti-association factor Tif6 from the large ribosomal subunit, which can consequently bind the small ribosomal subunit and enter translation.Les travaux de ce mémoire portent sur la dynamique d'assemblage, de dissociation et de recyclage des protéines impliquées dans la biogenèse de la grande sous-unité ribosomique chez la levure Saccharomyces cerevisiae. Ils permettent une meilleure compréhension de deux points de contrôle de cette voie métabolique, l'un dans le noyau, l'autre dans le cytoplasme. Nous avons montré que la protéine nucléaire Nsa2, extrêmement conservée chez les Eucaryotes, est requise pour la maturation correcte de l'intermédiaire d'ARN ribosomique 27SB. Nsa2 est un facteur instable et régulé en fonction de l'activité de la biogenèse des ribosomes ; à ce titre, il pourrait centraliser différents signaux de contrôle de la voie métabolique. Par ailleurs, la technique de SILAC nous a permis de définir des groupes de facteurs pré-ribosomiques précoces ou tardifs par rapport au point d'action de Nsa2.Dans le cytoplasme, nous avons mis en évidence un réseau de protéines marquant la transition entre la fin de la biogenèse de la grande sous-unité et l'initiation de la traduction. La protéine cytoplasmique Rei1 et la karyophérine Kap121 sont requises pour le recyclage du dimère de facteurs navettes Arx1-Alb1, du cytoplasme vers le noyau. Ce recyclage conditionne la dissociation entre le facteur d'anti-association Tif6 et la grande sous-unité ribosomique, qui peut dès lors se lier à la petite sous-unité ribosomique et participer à la traduction

La mise en évidence du bacille de la peste, Hong-Kong 1894

Bien qu’elle ait décimé des millions de personnes au cours des siècles, la peste ne constitue plus aujourd’hui un problème majeur de santé publique. Transmise par les puces, cette maladie affecte surtout les petits animaux, et peut occasionnellement être transmise à l’homme. Selon les chiffres de l’Organisation Mondiale de la Santé, le nombre de cas humains en 2003 était de 2118, dont 182 mortels.Comment cette maladie tristement célèbre est-elle passée, en un siècle, du rang de fléau de l’humanité à celui de pathologie rare et relativement bien contrôlée ? La description précise de la bactérie par le médecin franco-suisse Alexandre Yersin, a été à la base de ce progrès : elle sera nommée Yersinia pestis en son honneur. Suite à ses travaux, les recherches se concentrèrent sur les moyens de soigner les malades par des sérothérapies et l’administration d’antibiotiques, et de protéger de l’affection par la mise au point de vaccins. Si, grâce à ces mesures, la peste n’est donc plus à proprement parler un sujet d’actualité, la méthodologie qui a conduit Yersin à sa découverte est toujours pertinente lorsque les scientifiques sont confrontés à une nouvelle épidémie et cherchent à en identifier les causes. Ainsi, ces dernières années, le même type de démarche a-t-il été mis en place afin d’étudier des maladies émergentes comme le SIDA, le SRAS, le chikungunya ou encore le virus Ébola.Although the plague has claimed the life of millions of people over the centuries, it no longer poses a major risk to public health. Spread by fleas, the disease mainly infects small animals, and can occasionally be transmitted to humans. According to figures released by the World Health Organization, 2,118 cases of human plague were reported in 2003, of which 182 were fatal.How, in the space of a century, did this notorious disease go from being the scourge of humanity to a rare and relatively well-controlled pathology? The key to this advance was the detailed description of the bacteria by the Franco-Swiss doctor Alexandre Yersin: the bacteria was named Yersinia pestis in his honour. Following Yersin’s work, research was oriented towards antiserum treatment, the administration of antibiotics, and the prevention of infection through the development of vaccines.Thanks to these measures, the plague is no longer in the news. Yet the methodology that led Yersin to make his discovery remains relevant whenever scientists are confronted with a new epidemic and attempt to identify its causes. Thus, in recent years, the same type of approach has been used to study emerging diseases such as AIDS, SARS, Chikungunya and the Ebola virus

A functional network involved in the recycling of nucleocytoplasmic pre-60S factors

Eukaryotic pre-ribosomes go through cytoplasmic maturation steps before entering translation. The nucleocytoplasmic proteins participating in these late stages of maturation are reimported to the nucleus. In this study, we describe a functional network focused on Rei1/Ybr267w, a strictly cytoplasmic pre-60S factor indirectly involved in nuclear 27S pre-ribosomal RNA processing. In the absence of Rei1, the nuclear import of at least three other pre-60S factors is impaired. The accumulation in the cytoplasm of a small complex formed by the association of Arx1 with a novel factor, Alb1/Yjl122w, inhibits the release of the putative antiassociation factor Tif6 from the premature large ribosomal subunits and its recycling to the nucleus. We propose a model in which Rei1 is a key factor for the coordinated dissociation and recycling of the last pre-60S factors before newly synthesized large ribosomal subunits enter translation

60S ribosomal subunit assembly dynamics defined by semi-quantitative mass spectrometry of purified complexes

During the highly conserved process of eukaryotic ribosome formation, RNA follows a maturation path with well-defined, successive intermediates that dynamically associate with many pre-ribosomal proteins. A comprehensive description of the assembly process is still lacking. To obtain data on the timing and order of association of the different pre-ribosomal factors, a strategy consists in the use of pre-ribsomal particles isolated from mutants that block ribosome formation at different steps. Immunoblots, inherently limited to only a few factors, have been applied to evaluate the accumulation or decrease of pre-ribosomal intermediates under mutant conditions. For a global protein-level description of different 60S ribosomal subunit maturation intermediates in yeast, we have adapted a method of in vivo isotopic labelling and mass spectrometry to study pre-60S complexes isolated from strains in which rRNA processing was affected by individual depletion of five factors: Ebp2, Nog1, Nsa2, Nog2 or Pop3. We obtained quantitative data for 45 distinct pre-60S proteins and detected coordinated changes for over 30 pre-60S factors in the analysed mutants. These results led to the characterisation of the composition of early, intermediate and late pre-ribosomal complexes, specific for crucial maturation steps during 60S assembly in eukaryotes

A bacterial protein targets the BAHD1 chromatin complex to stimulate type III interferon response

International audienceIntracellular pathogens such as $Listeria\ monocytogenes$ subvert cellular functions through the interaction of bacterial effectors with host components. Here we found that a secreted listerial virulence factor, LntA, could target the chromatin repressor BAHD1 in the host cell nucleus to activate interferon IFN-stimulated genes (ISGs). IFN-$\lambda$ expression was induced in response to infection of epithelial cells with bacteria lacking LntA; however, the BAHD1-chromatin associated complex repressed downstream ISGs. In contrast, in cells infected with $lntA$-expressing bacteria, LntA prevented BAHD1 recruitment to ISGs and stimulated their expression. Murine listeriosis decreased in BAHD1$^{+/-}$ mice or when $lntA$ was constitutively expressed. Thus, the LntA-BAHD1 interplay may modulate IFN-$\lambda$-mediated immune response to control bacterial colonization of the host

Role of the BAHD1 Chromatin-Repressive Complex in Placental Development and Regulation of Steroid Metabolism.

BAHD1 is a vertebrate protein that promotes heterochromatin formation and gene repression in association with several epigenetic regulators. However, its physiological roles remain unknown. Here, we demonstrate that ablation of the Bahd1 gene results in hypocholesterolemia, hypoglycemia and decreased body fat in mice. It also causes placental growth restriction with a drop of trophoblast glycogen cells, a reduction of fetal weight and a high neonatal mortality rate. By intersecting transcriptome data from murine Bahd1 knockout (KO) placentas at stages E16.5 and E18.5 of gestation, Bahd1-KO embryonic fibroblasts, and human cells stably expressing BAHD1, we also show that changes in BAHD1 levels alter expression of steroid/lipid metabolism genes. Biochemical analysis of the BAHD1-associated multiprotein complex identifies MIER proteins as novel partners of BAHD1 and suggests that BAHD1-MIER interaction forms a hub for histone deacetylases and methyltransferases, chromatin readers and transcription factors. We further show that overexpression of BAHD1 leads to an increase of MIER1 enrichment on the inactive X chromosome (Xi). In addition, BAHD1 and MIER1/3 repress expression of the steroid hormone receptor genes ESR1 and PGR, both playing important roles in placental development and energy metabolism. Moreover, modulation of BAHD1 expression in HEK293 cells triggers epigenetic changes at the ESR1 locus. Together, these results identify BAHD1 as a core component of a chromatin-repressive complex regulating placental morphogenesis and body fat storage and suggest that its dysfunction may contribute to several human diseases

Exo-zodi Modeling for the Large Binocular Telescope Interferometer

Habitable zone dust levels are a key unknown that must be understood to ensure the success of future space missions to image Earth analogs around nearby stars. Current detection limits are several orders of magnitude above the level of the solar system's zodiacal cloud, so characterization of the brightness distribution of exo-zodi down to much fainter levels is needed. To this end, the Large Binocular Telescope Interferometer (LBTI) will detect thermal emission from habitable zone exo-zodi a few times brighter than solar system levels. Here we present a modeling framework for interpreting LBTI observations, which yields dust levels from detections and upper limits that are then converted into predictions and upper limits for the scattered light surface brightness. We apply this model to the HOSTS survey sample of nearby stars; assuming a null depth uncertainty of 10^(–4) the LBTI will be sensitive to dust a few times above the solar system level around Sun-like stars, and to even lower dust levels for more massive stars

An RNA-Binding Protein Secreted by a Bacterial Pathogen Modulates RIG-I Signaling.

RNA-binding proteins (RBPs) perform key cellular activities by controlling the function of bound RNAs. The widely held assumption that RBPs are strictly intracellular has been challenged by the discovery of secreted RBPs. However, extracellular RBPs have been described in eukaryotes, while secreted bacterial RBPs have not been reported. Here, we show that the bacterial pathogen Listeria monocytogenes secretes a small RBP that we named Zea. We show that Zea binds a subset of L. monocytogenes RNAs, causing their accumulation in the extracellular medium. Furthermore, during L. monocytogenes infection, Zea binds RIG-I, the non-self-RNA innate immunity sensor, potentiating interferon-β production. Mouse infection studies reveal that Zea affects L. monocytogenes virulence. Together, our results unveil that bacterial RNAs can be present extracellularly in association with RBPs, acting as "social RNAs" to trigger a host response during infection

La Cappadoce méridionale de la Préhistoire à l'époque byzantine

Il y a environ 25 ans, Olivier Pelon organisait à l’Institut Français d’Études Anatoliennes d’Istanbul un colloque destiné à faire l’état des recherches sur la Cappadoce méridionale jusqu’à la fin de l’époque romaine. Un quart de siècle après ce premier colloque, il était intéressant de faire un nouveau point sur l’avancée des recherches dans cette Cappadoce méridionale, de la préhistoire à la période byzantine. Ce nouveau colloque, placé cette fois encore sous l’égide de l’Institut Français d’Études Anatoliennes et intégré à la série des Rencontres d’archéologie de l’IFEA réunit vingt-trois communications. Si les périodes néolithique et chalcolithique ont été particulièrement bien représentées, ce qui témoigne bien de l’importance de cette phase de la préhistoire cappadocienne, liée aux gisements d’obsidienne des Melendiz Dağları, on soulignera en revanche l’absence presque totale du Bronze Ancien. Cette phase est en effet peu représentée dans l’archéologie locale. La même remarque peut s’appliquer au Bronze Moyen. La fin du Bronze Moyen, fort heureusement, est représentée à Porsuk, de même que le Bronze Récent qui bénéficie, depuis peu, tout comme l’Âge du Fer, du démarrage fructueux des fouilles de Kınık Höyük. Enfin, l’Antiquité tardive et Byzance ont pu être représentées, principalement autour de Tyane, ce qui n’avait pas pu être le cas lors du premier colloque. En octobre 2012, quelques semaines avant la tenue de la Rencontre, on apprenait malheureusement le décès brutal et inattendu d’Olivier Pelon, ancien directeur de la mission de Porsuk (jusqu’en 2002) et organisateur de ce premier colloque cappadocien. C’est bien en hommage à sa mémoire que notre Rencontre cappadocienne de 2012 et sa publication ont été naturellement dédiées

Socializing One Health: an innovative strategy to investigate social and behavioral risks of emerging viral threats

In an effort to strengthen global capacity to prevent, detect, and control infectious diseases in animals and people, the United States Agency for International Development’s (USAID) Emerging Pandemic Threats (EPT) PREDICT project funded development of regional, national, and local One Health capacities for early disease detection, rapid response, disease control, and risk reduction. From the outset, the EPT approach was inclusive of social science research methods designed to understand the contexts and behaviors of communities living and working at human-animal-environment interfaces considered high-risk for virus emergence. Using qualitative and quantitative approaches, PREDICT behavioral research aimed to identify and assess a range of socio-cultural behaviors that could be influential in zoonotic disease emergence, amplification, and transmission. This broad approach to behavioral risk characterization enabled us to identify and characterize human activities that could be linked to the transmission dynamics of new and emerging viruses. This paper provides a discussion of implementation of a social science approach within a zoonotic surveillance framework. We conducted in-depth ethnographic interviews and focus groups to better understand the individual- and community-level knowledge, attitudes, and practices that potentially put participants at risk for zoonotic disease transmission from the animals they live and work with, across 6 interface domains. When we asked highly-exposed individuals (ie. bushmeat hunters, wildlife or guano farmers) about the risk they perceived in their occupational activities, most did not perceive it to be risky, whether because it was normalized by years (or generations) of doing such an activity, or due to lack of information about potential risks. Integrating the social sciences allows investigations of the specific human activities that are hypothesized to drive disease emergence, amplification, and transmission, in order to better substantiate behavioral disease drivers, along with the social dimensions of infection and transmission dynamics. Understanding these dynamics is critical to achieving health security--the protection from threats to health-- which requires investments in both collective and individual health security. Involving behavioral sciences into zoonotic disease surveillance allowed us to push toward fuller community integration and engagement and toward dialogue and implementation of recommendations for disease prevention and improved health security