Etude de la maturation cytoplasmique de la petite sous-unité ribosomique chez Saccharomyces cerevisiae

Les ribosomes constituent un des acteurs majeurs du mécanisme de traduction dans toute cellule vivante. La synthèse des ribosomes est un processus complexe commençant par la transcription d'un pré-ARN ribosomique (ARNr) contenant les futurs ARNr matures ainsi que des séquences qui vont être coupées tout au long de la biogenèse des sous-unités ribosomiques. Chez Saccharomyces cerevisiae, ce ne sont pas moins de 200 facteurs qui interviennent tout au long de ce processus. Nous nous sommes intéressés plus précisément à l'étape cytoplasmique de maturation de la petite sous-unité ribosomique, consistant en une coupure endonucléolytique permettant de passer d'un pré-ARNr 20S contenu dans une particule pré-40S à un ARNr 18S mature contenu dans une sous-unité 40S. Cette sous-unité mature peut ensuite initier la traduction. Le modèle initial proposait que la maturation de la petite sous-unité fût un prérequis à l'initiation de la traduction. Nos expériences ont permis d'observer qu'une fraction du pré-ARNr 20S cosédimente avec les complexes de 80S et polysomes. Cette fraction de pré-ARNr 20S est augmentée dans certains mutants où l'on bloque la maturation de la petite-sous unité dans le cytoplasme. Nous avons confirmé l'existence de ribosomes contenant des particules pré-40S et interagissant avec des ARNm par des approches biochimiques. Ainsi, nos données suggèrent que des sous-unités ribosomiques non matures peuvent initier la traduction. Ces ribosomes aberrants sont alors dégradés via le No Go decay, un mécanisme de contrôle-qualité des ARNs cytoplasmiques. Ainsi, le No Go Decay fonctionnerait comme le mécanisme ultime de contrôle-qualité des sous-unités ribosomiques 40S.Ribosomes constitute one of the major actors of the mechanism of translation in any living cell. The synthesis of ribosomes is a complex process beginning with the transcription of a pre-ribosomal RNA (rRNA) containing future mature rRNAs as well as sequences that are eliminated during ribosome biogenesis. In Saccharomyces cerevisiae, no less than 200 factors are implicated in this process. We were more precisely interested in the cytoplasmic step of the small ribosomal subunit maturation consisting of an endonucleolytic cleavage of the 20S pre-rRNA contained in a pre-40S particle and leading to the mature 18S rRNA contained in the 40S ribosomal subunit. The initial model was that 40S ribosomal subunit maturation might be a pre-requisite for translation initiation. Our experiments have led to the observation that a fraction of 20S pre-rRNA co-sediments with 80S complexes and polysomes. This 20S pre-rRNA fraction can be increased in mutants impaired in the cytoplasmic step of 40S ribosomal subunit maturation. By biochemical approaches, we confirmed the occurrence of ribosomes containing pre-40S particles and mRNAs. Thus, our data suggest that pre-40S particles can initiate translation. These aberrant ribosomes are then degraded via the No Go decay pathway involved in the quality control of some cytoplasmic RNAs. No-Go Decay would function as an ultimate quality control mechanism of the 40S ribosomal subunit

Science with an electrodynamic tether

Ionospheric interaction experiments using a conductive, fully bare tether are discussed. With an optimal design, requiring 1.15 mm diameter and 7.5 km full length for a collected current of 0.87 A at day conditions, the tether radiates 0.33 watts as Fast Magnetosonic waves and 0.16 watts as Alfven waves. Secondary keV electrons are produced over a 6.5 km length, giving raise to noticeable auroral effects in the D-layer, at low geomagnetic latitudes. A preliminary design of the experiment, to be implemented on either a satellite or a Station, has been carried out. An ejector gives an initial velocity to an end mass, a free spool of tether unwinding from that mass during a first stage of deployment; other phases are monitored through the tether velocity, driving a reel with an unwinding device

Histone exchange is associated with activator function at transcribed promoters and with repression at histone loci

Transcription in eukaryotes correlates with major chromatin changes, including the replacement of old nucleosomal histones by new histones at the promoters of genes. The role of these histone exchange events in transcription remains unclear. In particular, the causal relationship between histone exchange and activator binding, preinitiation complex (PIC) assembly, and/or subsequent transcription remains unclear. Here, we provide evidence that histone exchange at gene promoters is not simply a consequence of PIC assembly or transcription but instead is mediated by activators. We further show that not all activators up-regulate gene expression by inducing histone turnover. Thus, histone exchange does not simply correlate with transcriptional activity, but instead reflects the mode of action of the activator. Last, we show that histone turnover is not only associated with activator function but also plays a role in transcriptional repression at the histone loci

Human RioK3 is a novel component of cytoplasmic pre-40S pre-ribosomal particles

Maturation of the 40S ribosomal subunit precursors in mammals mobilizes several non-ribosomal proteins, including the atypical protein kinase RioK2. Here, we have investigated the involvement of another member of the RIO kinase family, RioK3, in human ribosome biogenesis. RioK3 is a cytoplasmic protein that does not seem to shuttle between nucleus and cytoplasm via a Crm1-dependent mechanism as does RioK2 and which sediments with cytoplasmic 40S ribosomal particles in a sucrose gradient. When the small ribosomal subunit biogenesis is impaired by depletion of either rpS15, rpS19 or RioK2, a concomitant decrease in the amount of RioK3 is observed. Surprisingly, we observed a dramatic and specific increase in the levels of RioK3 when the biogenesis of the large ribosomal subunit is impaired. A fraction of RioK3 is associated with the non ribosomal pre-40S particle components hLtv1 and hEnp1 as well as with the 18S-E pre-rRNA indicating that it belongs to a bona fide cytoplasmic pre-40S particle. Finally, RioK3 depletion leads to an increase in the levels of the 21S rRNA precursor in the 18S rRNA production pathway. Altogether, our results strongly suggest that RioK3 is a novel cytoplasmic component of pre-40S pre-ribosomal particle(s) in human cells, required for normal processing of the 21S pre-rRNA

Annexin-V positive extracellular vesicles level is increased in severe COVID-19 disease

ObjectivesTo evaluate extracellular vesicles levels in a cohort of SARS-CoV-2’s patients hospitalized in an intensive care unit with and without COVID-19 associated thromboembolic events.MethodsIn this study, we aim to assess endothelial and platelet membrane-derived extracellular vesicles levels in a cohort of SARS-CoV-2 patients with and without COVID-19-associated thromboembolic events who were hospitalized in an intensive care unit. Annexin-V positive extracellular vesicles levels were prospectively assessed by flow cytometry in one hundred twenty-three critically ill adults diagnosed with acute respiratory distress syndrome associated with a SARS-CoV-2 infection, ten adults diagnosed for moderate SARS-CoV-2 infection and 25 healthy volunteers.ResultsOn our critically ill patients, thirty-four patients (27.6%) had a thromboembolic event, Fifty-three (43%) died. Endothelial and platelet membrane-derived extracellular vesicles were drastically increased in SARS-CoV-2 patients hospitalized in the ICU compared to healthy volunteers. Moreover a slighty higher small/large ratio for platelets membrane-derived extracellular vesicles in patients was linked to thrombo-embolic events.ConclusionA comparison between total annexin-V positive extracellular vesicles levels in severe and moderate SARS-CoV-2 infection and healthy controls showed a significant increase in patients with severe infection and their sizes could be considered as biomarkers of SARS-CoV-2 associated thrombo-embolic events

Nuclear DNA Replication in Trypanosomatids:There Are No Easy Methods for Solving Difficult Problems

In trypanosomatids, etiological agents of devastating diseases, replication is robust and finely controlled to maintain genome stability and function in stressful environments. However, these parasites encode several replication protein components and complexes that show potentially variant composition compared with model eukaryotes. This review focuses on the advances made in recent years regarding the differences and peculiarities of the replication machinery in trypanosomatids, including how such divergence might affect DNA replication dynamics and the replication stress response. Comparing the DNA replication machinery and processes of parasites and their hosts may provide a foundation for the identification of targets that can be used in the development of chemotherapies to assist in the eradication of diseases caused by these pathogens

Proofreading of pre-40S ribosome maturation by a translation initiation factor and 60S subunits

In the final steps of yeast ribosome synthesis, immature translation-incompetent pre-40S particles that contain 20S pre-rRNA are converted to the mature translation-competent subunits containing the 18S rRNA. An assay for 20S pre-rRNA cleavage in purified pre-40S particles showed that cleavage by the PIN domain endonuclease Nob1 was strongly stimulated by the GTPase activity of the cytoplasmic translation initiation factor eIF5b/Fun12. Cleavage of the 20S pre-rRNA was also inhibited in vivo and in vitro by blocking binding of Fun12 to the 25S rRNA through specific methylation of its binding site. Cleavage competent pre-40S particles stably associate with Fun12 and form 80S complexes with 60S ribosomal subunits. We propose that recruitment of 60S subunits promotes GTP-hydrolysis by Fun12, leading to structural rearrangements within the pre-40S particle that bring Nob1 and the pre-rRNA cleavage site together

Final Pre-40S Maturation Depends on the Functional Integrity of the 60S Subunit Ribosomal Protein L3

Ribosomal protein L3 is an evolutionarily conserved protein that participates in the assembly of early pre-60S particles. We report that the rpl3[W255C] allele, which affects the affinity and function of translation elongation factors, impairs cytoplasmic maturation of 20S pre-rRNA. This was not seen for other mutations in or depletion of L3 or other 60S ribosomal proteins. Surprisingly, pre-40S particles containing 20S pre-rRNA form translation-competent 80S ribosomes, and translation inhibition partially suppresses 20S pre-rRNA accumulation. The GTP-dependent translation initiation factor Fun12 (yeast eIF5B) shows similar in vivo binding to ribosomal particles from wild-type and rpl3[W255C] cells. However, the GTPase activity of eIF5B failed to stimulate processing of 20S pre-rRNA when assayed with ribosomal particles purified from rpl3[W255C] cells. We conclude that L3 plays an important role in the function of eIF5B in stimulating 3′ end processing of 18S rRNA in the context of 80S ribosomes that have not yet engaged in translation. These findings indicate that the correct conformation of the GTPase activation region is assessed in a quality control step during maturation of cytoplasmic pre-ribosomal particles

Bonnaud-Delamare (Roger) - Attributions juridiques des préfets et sous-préfets

Soudet P. Bonnaud-Delamare (Roger) - Attributions juridiques des préfets et sous-préfets. In: Revue française de science politique, 3ᵉ année, n°2, 1953. pp. 405-406