Insertion and translocation of proteins into and through membranes

AbstractIn prokaryotic and eukaryotic organisms proteins are efficiently sorted to reach their final destinations in a whole range of subcellular compartments. Targeting is mediated by Hydrophobie signal sequences or hydrophilic targeting sequences depending upon the compartment, these sequences being often processed. Proteins cannot be translocated through a membrane in a tightly folded stage, they must have a loose conformation, the so-called ‘translocation competent state’, which is usually kept through interactions with chaperones. In addition to these cytosolic receptor-like components, receptors are also present on the target membranes. Depending upon the organelles and organisms, two different energy sources have been identified, energy rich phosphate bonds (ATP and GTP) and a potential across the target membrane. Besides the signal peptides various classes of signals have been identified to account for topologies of membrane proteins. Protein secretion in bacterial organisms has been extensively studied. Various classes of proteins use different strategies some of these may also be used in eukaryotic cells

Search for dark matter produced in association with bottom or top quarks in √s = 13 TeV pp collisions with the ATLAS detector

A search for weakly interacting massive particle dark matter produced in association with bottom or top quarks is presented. Final states containing third-generation quarks and miss- ing transverse momentum are considered. The analysis uses 36.1 fb−1 of proton–proton collision data recorded by the ATLAS experiment at √s = 13 TeV in 2015 and 2016. No significant excess of events above the estimated backgrounds is observed. The results are in- terpreted in the framework of simplified models of spin-0 dark-matter mediators. For colour- neutral spin-0 mediators produced in association with top quarks and decaying into a pair of dark-matter particles, mediator masses below 50 GeV are excluded assuming a dark-matter candidate mass of 1 GeV and unitary couplings. For scalar and pseudoscalar mediators produced in association with bottom quarks, the search sets limits on the production cross- section of 300 times the predicted rate for mediators with masses between 10 and 50 GeV and assuming a dark-matter mass of 1 GeV and unitary coupling. Constraints on colour- charged scalar simplified models are also presented. Assuming a dark-matter particle mass of 35 GeV, mediator particles with mass below 1.1 TeV are excluded for couplings yielding a dark-matter relic density consistent with measurements

Etude des protéines de la famille des PEBP dans la levure Saccharomyces cerevisiae (mise en évidence d'un nouvel activateur de la voie Ras-AMPc-PKA)

Les petites protéines G Ras sont des protéines très conservées impliquées dans des mécanismes de transduction du signal et régulées par deux types de protéines antagonistes: des GTPase-activating proteins (GAPs) et des guanine exchange proteins (GEFs). Ce travail décrit le premier inhibiteur physiologique de Ira2, une protéine GAP de Ras chez la levure Saccharomyces cerevisiae : la protéine Tfs1. TFS1, décrit comme un suppresseur multicopies de cdc25.1, interagit spécifiquement avec Ira2 et l inhibe in vivo. Tfs1 fait partie, comme son homologue de fonction inconnue Ylr179c, de la famille des PEBP (Phosphatidylethanolamine binding protein). La cavité conservée de cette famille joue un rôle important pour l interaction Tfs1/Ira2. Un modèle moléculaire de la structure 3D de la protéine Tfs1 a été construit. Des protéines hybrides entre Tfs1 et Ylr179c ont également été réalisées et leur fonction a été testée et interprétée au point de vue structural.ORLEANS-BU Sciences (452342104) / SudocSudocFranceF

Etude de la régulation d'une protéine GAP de Ras de la levure à l'homme

La neurofibromatose de type 1 est une maladie génétique fréquente puisqu elle touche 1 individu sur 3500. Le gène responsable de la maladie a été identifié et code pour une protéine, la neurofibromine (Nf1), exprimée de manière ubiquitaire mais plus abondamment dans les neurones, les astrocytes, les cellules de Schwann et les oligodendrocytes. Il est un enjeu de taille de comprendre la régulation de cette protéine.Une étude modèle a été développée chez la levure afin de caractériser les bases moléculaires de l interaction entre l homologue de Nf1, Ira2p, et une protéine qui l inhibe, Tfs1p. Cette étude a permis d identifier les régions de Tfs1p importantes pour l interaction à savoir l extrémité N-terminale, la cavité de surface, ainsi qu une région électropositive contenant ces deux déterminants. Nous nous sommes intéressés ensuite au rôle physiologique de Tfs1p dans les cellules. Des études systématiques ont suggéré un rôle prédominant de l inhibition d Ira2p dans des conditions de surproduction de Tfs1p. Ce rôle a été précisé et nous avons montré que Tfs1p participait, via cette inhibition, à la boucle de rétrocontrôle négatif de la réponse au stress.Sur la base de cette étude, nous avons cherché à savoir si l interaction Ira2p/Tfs1p était conservée chez l homme entre Nf1 et RKIP, homologue de Tfs1p. Nous avons en parallèle développé un crible double hybride sur Nf1 dans une banque d ADNc de cerveau humain afin de mettre en évidence de nouveaux régulateurs de ses fonctions. 1464 candidats positifs ont été isolés. Actuellement, sur 8% de candidats identifiés 3 partenaires intéressants ont déjà pu être mis en évidence.ORLEANS-BU Sciences (452342104) / SudocSudocFranceF

LIMK2-1, a new isoform of human LIMK2, regulates actin cytoskeleton remodeling via a different signaling pathway than that of its two homologs, LIMK2a and LIMK2b

International audienceLIMK1 and LIMK2 (LIMKs) are kinases that play a crucial role in cytoskeleton dynamics by independently regulating both actin filament and microtubule remodeling. LIMK1, and more recently LIMK2, have been shown to be involved in cancer development and metastasis, resistance of cancer cells to microtubule targeted treatments, neurological diseases, and viral infection. LIMKs have thus recently emerged as new therapeutic targets. Databanks describe three isoforms of human LIMK2: LIMK2a, LIMK2b, and LIMK2-1. Evidence suggests that they may not have completely overlapping functions. We biochemically characterized the three isoforms to better delineate their potential roles, focusing on LIMK2-1, which has only been described at the mRNA level in a single study. LIMK2-1 has a Protein Phosphatase 1 (PP1) inhibitory domain at its C-terminus which its two counterparts do not. We showed that the LIMK2-1 protein is indeed synthesized. LIMK2-1 does not phosphorylate cofilin, the canonical substrate of LIMKs, although it has kinase activity and promotes actin stress fiber formation. Instead, it interacts with PP1 and partially inhibits its activity towards cofilin. Our data suggest that LIMK2-1 regulates actin cytoskeleton dynamics by preventing PP1-mediated cofilin dephosphorylation, rather than by directly phosphorylating cofilin as its two counterparts, LIMK2a and LIMK2b. This specificity may allow for tight regulation of the phospho-cofilin pool, determining the fate of the cell

Assessing CeO2 and TiO2 Nanoparticle Concentrations in the Seine River and Its Tributaries Near Paris

International audienceMotivation for detecting engineered nanoparticles (ENPs) in the environment comes from a need to understand fate and behavior of these materials in natural matrices. The difficulty lies in the low expected ENP particle number concentration (PNC) and the presence of a large and variable background concentration of natural NPs. We report the PNCs and characteristics of cerium-bearing nanoparticles (Ce-NPs) and titanium-bearing nanoparticles (Ti-NPs) in an aquatic matrix (the Seine River and three of its tributaries) with the use of single particle ICP-MS (spICPMS) and electron microscopy (FEG-SEM). Ce-bearing and Ti-bearing particles were observed in suspended particulate matter collected onto 0.2 μm and 1 kDa filters, using FEG-SEM imaging. At Marnay-sur-Seine, the upstream point, PNCs for Ce-NPs and Ti-NPs were 0.47 ± 0.07 × 106 and 1.35 ± 0.17 × 106 particles as measured by spICPMS. The maximum PNC for both Ce-NPs and Ti-NPs, 1.59 ± 0.10 × 106 particles mL−1 and 5.89 ± 0.10 × 106 particles mL−1, respectively, were found in the Marne River, a major tributary to the Seine. It was shown that downstream of each confluence, an increase in the PNC of the Seine is observed, suggesting a significant contribution of the different tributaries. Mass balance of particles flows and elemental ratios of Ce/La showed that in the Marne and the Oise River, a contribution of natural CeO2 NPs exists. The anthropogenic contribution in TiO2 ENPs for the Marne River was further assessed with Ti/Al, Ti/V, and Ti/Y elemental ratios. Near constant element ratios in the Seine below the Orge River and Paris city suggest neither contribute significantly to Ce or Ti NP concentrations. The study provides further investigation of the strengths and limitations of the application of spICPMS to natural samples and contributes data to the currently highly-limited dataset on natural NP backgrounds in rivers, information that is key to assessing the potential for quantifying the input of ENPs to surface waters. Of the total mass of Ce and Ti, 83 and 90%, respectively, could be detected as particles by spICPMS

Datations croisées des occupations humaines et animales dans la grotte Chauvet Pont d'Arc

National audienc

LIMK2-1 is a Hominidae-Specific Isoform of LIMK2 Expressed in Central Nervous System and Associated with Intellectual Disability

International audienceLIMK2 is involved in neuronal functions by regulating actin dynamics. Different isoforms of LIMK2 are described in databanks. LIMK2a and LIMK2b are the most characterized. A few pieces of evidence suggest that LIMK2 isoforms might not have overlapping functions. In this study, we focused our attention on a less studied human LIMK2 isoform, LIMK2-1. Compared to the other LIMK2 isoforms, LIMK2-1 contains a supplementary C-terminal phosphatase 1 inhibitory domain (PP1i). We found out that this isoform was hominidae-specific and showed that it was expressed in human fetal brain and faintly in adult brain. Its coding sequence was sequenced in 173 patients with sporadic non-syndromic intellectual disability (ID), and we observed an association of a rare missense variant in the PP1i domain (rs151191437, p.S668P) with ID. Our results also suggest an implication of LIMK2-1 in neurite outgrowth and neurons arborization which appears to be affected by the p.S668P variation. Therefore our results suggest that LIMK2-1 plays a role in the developing brain, and that a rare variation of this isoform is a susceptibility factor in ID

Both ChIP-SEQ and in planta gene modification demonstrate the involvement of MYB090 and MYB156 in secondary cell wall formation in poplar

Lakhal W., Boizot N., Lesage-Descauses M-C., Rogier O., Lainé-Prade V., Laurans F., Ader K., Charpentier J-P., Decoville M., Bénédetti H., Leplé J-C., Pilate G., Déjardin A, 2015. Both ChIP-SEQ and in planta gene modification demonstrate the involvement of MYB090 and MYB156 in secondary cell wall formation in poplar. INUPRAG Meeting, 6-8 October 2015, Nancy, France, communication orale.Lakhal W., Boizot N., Lesage-Descauses M-C., Rogier O., Lainé-Prade V., Laurans F., Ader K., Charpentier J-P., Decoville M., Bénédetti H., Leplé J-C., Pilate G., Déjardin A, 2015. Both ChIP-SEQ and in planta gene modification demonstrate the involvement of MYB090 and MYB156 in secondary cell wall formation in poplar. INUPRAG Meeting, 6-8 October 2015, Nancy, France, communication orale.Both ChIP-SEQ and in planta gene modification demonstrate the involvement of MYB090 and MYB156 in secondary cell wall formation in poplar. INUPRAG-Meeting 201