The Stargazin-Related Protein {gamma}7 Interacts with the mRNA-Binding Protein Heterogeneous Nuclear Ribonucleoprotein A2 and Regulates the Stability of Specific mRNAs, Including CaV2.2

The role(s) of the novel stargazin-like {gamma}-subunit proteins remain controversial. We have shown previously that the neuron-specific {gamma}7 suppresses the expression of certain calcium channels, particularly CaV2.2, and is therefore unlikely to operate as a calcium channel subunit. We now show that the effect of {gamma}7 on CaV2.2 expression is via an increase in the degradation rate of CaV2.2 mRNA and hence a reduction of CaV2.2 protein level. Furthermore, exogenous expression of {gamma}7 in PC12 cells also decreased the endogenous CaV2.2 mRNA level. Conversely, knockdown of endogenous {gamma}7 with short-hairpin RNAs produced a reciprocal enhancement of CaV2.2 mRNA stability and an increase in endogenous calcium currents in PC12 cells. Moreover, both endogenous and expressed {gamma}7 are present on intracellular membranes, rather than the plasma membrane. The cytoplasmic C terminus of {gamma}7 is essential for all its effects, and we show that {gamma}7 binds directly via its C terminus to a heterogeneous nuclear ribonucleoprotein (hnRNP A2), which also binds to a motif in CaV2.2 mRNA, and is associated with native CaV2.2 mRNA in PC12 cells. The expression of hnRNP A2 enhances CaV2.2 IBa, and this enhancement is prevented by a concentration of {gamma}7 that alone has no effect on IBa. The effect of {gamma}7 is selective for certain mRNAs because it had no effect on {alpha}2{delta}-2 mRNA stability, but it decreased the mRNA stability for the potassium-chloride cotransporter, KCC1, which contains a similar hnRNP A2 binding motif to that in CaV2.2 mRNA. Our results indicate that {gamma}7 plays a role in stabilizing CaV2.2 mRNA

Determinants of the voltage dependence of G protein modulation within calcium channel β subunits

CaVβ subunits of voltage-gated calcium channels contain two conserved domains, a src-homology-3 (SH3) domain and a guanylate kinase-like (GK) domain with an intervening HOOK domain. We have shown in a previous study that, although Gβγ-mediated inhibitory modulation of CaV2.2 channels did not require the interaction of a CaVβ subunit with the CaVα1 subunit, when such interaction was prevented by a mutation in the α1 subunit, G protein modulation could not be removed by a large depolarization and showed voltage-independent properties (Leroy et al., J Neurosci 25:6984–6996, 2005). In this study, we have investigated the ability of mutant and truncated CaVβ subunits to support voltage-dependent G protein modulation in order to determine the minimal domain of the CaVβ subunit that is required for this process. We have coexpressed the CaVβ subunit constructs with CaV2.2 and α2δ-2, studied modulation by the activation of the dopamine D2 receptor, and also examined basal tonic modulation. Our main finding is that the CaVβ subunit GK domains, from either β1b or β2, are sufficient to restore voltage dependence to G protein modulation. We also found that the removal of the variable HOOK region from β2a promotes tonic voltage-dependent G protein modulation. We propose that the absence of the HOOK region enhances Gβγ binding affinity, leading to greater tonic modulation by basal levels of Gβγ. This tonic modulation requires the presence of an SH3 domain, as tonic modulation is not supported by any of the CaVβ subunit GK domains alone

Electronic Structure and Solvation Effects from Core and Valence Photoelectron Spectroscopy of Serum Albumin

International audienceX-ray photoelectron spectroscopy of bovine serum albumin (BSA) in a liquid jet is used to investigate the electronic structure of a solvated protein, yielding insight into charge transfer mechanisms in biological systems in their natural environment. No structural damage was observed in BSA following X-ray photoelectron spectroscopy in a liquid jet sample environment. Carbon and nitrogen atoms in different chemical environments were resolved in the X-ray photoelectron spectra of both solid and solvated BSA. The calculations of charge distributions demonstrate the difficulty of assigning chemical contributions in complex systems in an aqueous environment. The high-resolution X-ray core electron spectra recorded are unchanged upon solvation. A comparison of the valence bands of BSA in both phases is also presented. These bands display a higher sensitivity to solvation effects. The ionization energy of the solvated BSA is determined at 5.7 ± 0.3 eV. Experimental results are compared with theoretical calculations to distinguish the contributions of various molecular components to the electronic structure. This comparison points towards the role of water in hole delocalization in proteins

Serum pepsinogens can help to discriminate between H. pylori-induced and auto-immune atrophic gastritis: Results from a prospective multicenter study

International audienceBackground: Serum pepsinogen (PG) testing is recommended by the European guidelines for diagnosis of chronic atrophic gastritis (CAG). However, wide variations in diagnostic performances are observed, due to the differences in the extent of gastric atrophy, and possibly in its origin (Helicobacter pylori-, autoimmune (AIG)). Aim. To analyze the diagnostic performances of PGs testing according to these different parameters, using enzyme-linked-immunosorbent serologic assay (ELISA) and chemiluminescent immunoassay (CLEIA). Methods: Serum samples from patients having undergone gastroscopy with biopsies in five French centers were collected prospectively. Sensitivity (Se), specificity (Sp), and Area Under Curve were analyzed according to the extent and origin of CAG. Results: Overall, 344 patients (156 males [45%]; mean age 58.8 [±14.2] years) were included, among whom 44 had AIG. Diagnostic performances of PG I for the detection of corpus CAG were excellent, with Se and Sp of 92.7% and 99.1% for ELISA and 90.5% and 98.2% for CLEIA, respectively. For AIG, corresponding values were 97.7% and 97.4% for ELISA, and 95.6% and 97.1% for CLEIA. In multivariate analysis, PG levels were associated with the auto-immune origin (p<0.001) but not with the extent of the atrophic gastritis. Conclusions: Pepsinogens are highly efficient for the diagnosis of corpus-limited CAG and allow to discriminate AIG from H. pylori-induced gastritis

An Observing Campaign of the Mutual Events Within (617) Patroclus-Menoetius Binary Trojan System

International audienceIn 2006-2007, the binary Trojan system (617) Patroclus-Menotius reached one of its annual equinoxes. As a consequence, the system underwent a 6 months season of mutual eclipses and occultations. We organized a campaign of observations of these mutual events mostly centered along the first semester of 2007. We took advantage of an orbit solution of the similary-sized binary system published by Marchis et al. (Nature, 2006) to predict the timing of mutual eclipses and occultations observable from January to July 2007. During the campaign, the magnitude of Patroclus system varied from 15.8 to 16.6 and its solar phase from 9°.5 to 2°.7 at opposition (end of March). The amplitude of the events ranged between 0.2 and 0.3 magnitude. A large number of stations around the world were involved in the campaign and 20 lightcurves with mutual events signature were collected (http://www.imcce.fr/page.php?nav=en/observateur/campagnes_obs/patroclus/). With such favorable circumstances, photometric observations of the events will provide tight constraints regarding physical properties of the system such as sizes, shapes, sidereal spin period and surface composition. Combining AO observations collected at Keck and Gemini with this lightcurve data, we have refined the orbital parameters of the binary system to an unprecedented accuracy. We will present these findings, the determination of the sizes and shapes, and other results from the data analysis. This material is partly based upon work supported by the National Aeronautics and Space Administration issue through the Science Mission Directorate Research and Analysis Programs number NNG05GF09G

An Observing Campaign of the Mutual Events Within (617) Patroclus-Menoetius Binary Trojan System

International audienceIn 2006-2007, the binary Trojan system (617) Patroclus-Menotius reached one of its annual equinoxes. As a consequence, the system underwent a 6 months season of mutual eclipses and occultations. We organized a campaign of observations of these mutual events mostly centered along the first semester of 2007. We took advantage of an orbit solution of the similary-sized binary system published by Marchis et al. (Nature, 2006) to predict the timing of mutual eclipses and occultations observable from January to July 2007. During the campaign, the magnitude of Patroclus system varied from 15.8 to 16.6 and its solar phase from 9°.5 to 2°.7 at opposition (end of March). The amplitude of the events ranged between 0.2 and 0.3 magnitude. A large number of stations around the world were involved in the campaign and 20 lightcurves with mutual events signature were collected (http://www.imcce.fr/page.php?nav=en/observateur/campagnes_obs/patroclus/). With such favorable circumstances, photometric observations of the events will provide tight constraints regarding physical properties of the system such as sizes, shapes, sidereal spin period and surface composition. Combining AO observations collected at Keck and Gemini with this lightcurve data, we have refined the orbital parameters of the binary system to an unprecedented accuracy. We will present these findings, the determination of the sizes and shapes, and other results from the data analysis. This material is partly based upon work supported by the National Aeronautics and Space Administration issue through the Science Mission Directorate Research and Analysis Programs number NNG05GF09G

Amélioration de la qualité des stations permanentes françaises à large bande possédant des installations à forage peu profond

International audienceWithin the framework of the Résif (French seismological and geodetic network) research infrastructure, a major extension of the French permanent broadband network (RLBP) has been initiated in 2011 with the objective to reach ~190 stations by 2020. This network will allow a more homogeneous covering of metropolitan France, encompassing a wide variety of tectonic and geological units. Presently the RLBP network consists in 124 operating stations.For each new potential site a test was conducted for ~3 weeks, usually burying the broadband sensor at 50-80 cm depth directly in the ground or within a light temporary vault.For the final set-up, we tested various hosting infrastructures at three prototype sites and decided to promote the installation of a posthole sensor in a steel-cased shallow borehole, 5-20m deep, for all the sites in open areas. 43 stations have already been installed this way and have been operated for 5 to 20 months.Résif is a national research infrastructure dedicated to the observation and understanding of the structure and dynamics of the Internal Earth. Résif is based on high-tech observation networks, composed of seismological, geodetic and gravimetric instruments deployed in a dense manner throughout France. The data collected make it possible to study with high spatial and temporal resolution the deformation of the ground, surface and deep structures, seismicity on a local and global scale and natural hazards, and more particularly seismic events, on French territory. Résif is part of the European (EPOS - European Plate Observing System) and global systems of instruments used to image the Earth's interior as a whole and to study many natural phenomena.Dans le cadre des infrastructures de recherche du Résif (Réseau sismologique et géodésique français), une extension majeure du réseau haut débit permanent français (RLBP) a été initiée en 2011 avec pour objectif d'atteindre ~190 stations en 2020. Ce réseau permettra une couverture plus homogène de la France métropolitaine, englobant une grande variété d'unités tectoniques et géologiques. Actuellement, le réseau RLBP compte 124 stations opérationnelles.Pour chaque nouveau site potentiel, un test a été effectué pendant environ 3 semaines, généralement en enterrant le capteur large bande à une profondeur de 50 à 80 cm directement dans le sol ou dans une voûte temporaire légère.Pour la mise en place finale, nous avons testé différentes infrastructures d'hébergement sur trois sites prototypes et avons décidé de promouvoir l'installation d'un capteur dans un puits de forage peu profond, 5-20m de profondeur, en acier, pour tous les sites en zones ouvertes. 43 stations ont déjà été installées de cette façon et ont été exploitées pendant 5 à 20 mois.Résif est une infrastructure de recherche nationale dédiée à l’observation et la compréhension de la structure et de la dynamique Terre interne. Résif se base sur des réseaux d’observation de haut niveau technologique, composés d’instruments sismologiques, géodésiques et gravimétriques déployés de manière dense sur tout le territoire français. Les données recueillies permettent d’étudier avec une haute résolution spatio-temporelle la déformation du sol, les structures superficielles et profondes, la sismicité à l’échelle locale et globale et les aléas naturels, et plus particulièrement sismiques, sur le territoire français. Résif s’intègre aux dispositifs européens (EPOS - European Plate Observing System) et mondiaux d’instruments permettant d’imager l’intérieur de la Terre dans sa globalité et d’étudier de nombreux phénomènes naturels