Les protéines FXYD : nouveaux régulateurs de la Na,K-ATPase

Les protéines FXYD appartiennent à une famille de petites protéines membranaires. Des études récentes suggèrent que six des sept membres de cette famille, FXYD1 (phospholemman), FXYD2 (sous-unité γ de la Na,K-ATPase), FXYD3 (Mat-8), FXYD4 (CHIF), FXYD5 (Ric) et FXYD7, sont des sous-unités auxiliaires de la Na, K-ATPase régulant son activité de manière tissu et isoforme spécifique. Ces résultats soulignent la complexité de la régulation des ions Na+ et K+ par la Na,K-ATPase qui est nécessaire pour assurer les fonctions propres de différents tissus comme la réabsorption du Na+ par le rein, la contraction musculaire et l’excitabilité neuronale. De plus, une mutation dans FXYD2 a été liée à certains cas d’hypomagnésémie, suggérant que des perturbations de la régulation de la Na,K-ATPase par les protéines FXYD seraient impliquées dans des états physiopathologiques. Une meilleure compréhension de ce nouveau mécanisme de régulation de la Na,K-ATPase pourrait nous aider à mieux comprendre son rôle dans les états physiopathologiques. Dans cet article, nous discutons les données les plus récentes sur le rôle des protéines FXYD dans la modulation de la Na, K-ATPase.Members of the FXYD protein family are small membrane proteins which are characterized by an FXYD motif, two conserved glycines and a serine residue. FXYD proteins show a tissue-specific distribution. Recent evidence suggests that 6 out of 7 FXYD proteins, FXYD1 (phospholemman), FXYD2 (γ subunit of Na,K-ATPase), FXYD3 (Mat-8), FXYD4 (CHIF), FXYD5 (Ric) and FXYD7 associate with Na,K-ATPase and modulate its transport properties e.g. its Na+ and/or its K+ affinity in a distinct way. These results highlight the complex regulation of Na+ and K+ transport which is necessary to ensure proper tissue functions such as renal Na+-reabsorption, muscle contractility and neuronal excitability. Moreover, mutation of a conserved glycine residue into an arginine residue in FXYD2 has been linked to cases of human hypomagnesemia indicating that dysregulation of Na,K-ATPase by FXYD proteins may be implicated in pathophysiological states. A better characterization of this novel regulatory mechanism of Na,K-ATPase may help to better understand its role in physiological and pathophysiological conditions

A novel family of transmembrane proteins interacting with β subunits of the Na,K-ATPase

We characterized a family consisting of four mammalian proteins of unknown function (NKAIN1, 2, 3 and 4) and a single Drosophila ortholog dNKAIN. Aside from highly conserved transmembrane domains, NKAIN proteins contain no characterized functional domains. Striking amino acid conservation in the first two transmembrane domains suggests that these proteins are likely to function within the membrane bilayer. NKAIN family members are neuronally expressed in multiple regions of the mouse brain, although their expression is not ubiquitous. We demonstrate that mouse NKAIN1 interacts with the β1 subunit of the Na,K-ATPase, whereas Drosophila ortholog dNKAIN interacts with Nrv2.2, a Drosophila homolog of the Na,K-ATPase β subunits. We also show that NKAIN1 can form a complex with another β subunit-binding protein, MONaKA, when binding to the β1 subunit of the Na,K-ATPase. Our results suggest that a complex between mammalian NKAIN1 and MONaKA is required for NKAIN function, which is carried out by a single protein, dNKAIN, in Drosophila. This hypothesis is supported by the fact that dNKAIN, but not NKAIN1, induces voltage-independent amiloride-insensitive Na+-specific conductance that can be blocked by lanthanum. Drosophila mutants with decreased dNKAIN expression due to a P-element insertion in the dNKAIN gene exhibit temperature-sensitive paralysis, a phenotype also caused by mutations in the Na,K-ATPase α subunit and several ion channels. The neuronal expression of NKAIN proteins, their membrane localization and the temperature-sensitive paralysis of NKAIN Drosophila mutants strongly suggest that this novel protein family may be critical for neuronal functio

Vitrine ou miroir de la nature du régime ? Le pavillon de l’Allemagne à l’Exposition universelle de 1937

Le succès trouvé par le pavillon allemand à l’Exposition universelle de 1937 ne doit pas faire illusion sur la réalité de la contribution allemande à cet événement international. Le contraste entre l’aspiration de l’Allemagne à briller de ses feux d’une part et les arcanes de sa participation d’autre part offre une intensité inattendue. Non seulement l’Allemagne, qui aspire à se positionner si fièrement sur la scène internationale, souffre d’une fragilité financière qui n’est pas loin de lui coûter sa participation. Mais, bien plus fondamentalement, l’étude de la participation allemande révèle que les principes idéologiques du IIIe Reich affectent, dès 1936-1937, bien au-delà de simples considérations liées à l’entreprise de propagande, les processus de la prise de décision de l’appareil administratif allemand, jusque, par exemple, dans le contrôle des ouvriers participant au chantier du pavillon et dans la gestion des flux touristiques à destination de l’Exposition internationale de Paris.Der Glanz und der Erfolg des deutschen Pavillons bei der Weltausstellung von 1937 sollen nicht über die Teilnahme Deutschlands am Ereignis. Der Kontrast zwischen dem Wunsch Deutschlands zur Geltung zu kommen, und den verborgenen Abläufen hinter den Kulissen des deutschen Beitrages, hätte kaum größer sein können. Das Deutschland, das sich mit seinem monumentalen Pavillon so stolz auf der internationalen Bühne zu positioniert versuchte, litt an einer dramatischen Finanzschwäche, die ihm fast die Teilnahme an der Weltausstellung gekostet hätte. Außerdem war die deutsche Teilnahme, weit über die propagandistischen Bestrebungen des Regimes hinaus, von den Paradigmen des Nationalsozialismus geprägt. Letztere waren sehr tiefgreifend und betrafen unter anderem den Umgang mit den am Bau beteiligten Arbeitern, aber auch den Reiseverkehr aus Deutschland zu der Weltausstellung in Paris.The positive response garnered by the pavilion of Nazi Germany at the world fair of 1937 can be misleading in regard to the circumstances of Germany’s participation to this international event. The contrast between Germany’s aspiration to show itself to its fullest advantage and, on the other hand, the facts about the German contribution, is unexpectedly intense. Germany, while it tried to present itself proudly on the international stage, suffered from a precarious financial situation that came close to costing it its participation. As early as 1936-1937, the national-socialist paradigms were also profoundly determining the process flow and decision making at play behind Germany’s participation in the event. These paradigms were far more pervasive than merely propagandistic endeavours, affecting, for instance, the management of the workforce involved in the construction of the German pavilion, as well as the touristic flows from Germany to France

Establishment of cell-cell junctions depends on the oligomeric states of VE-cadherin.: Oligomerization of VE-cadherin at cell surface

International audienceSpecifically expressed at intercellular adherens junctions of endothelial cells, VE-cadherin is a receptor that exhibits particular self-association properties. Indeed, in vitro studies demonstrated that the extracellular part of VE-cadherin elaborates Ca(++)-dependent hexameric structures. We hypothesized that this assembly could be at the basis of a new cadherin-mediated cell-cell adhesion mechanism. To verify this assumption, we first demonstrated that VE-cadherin can elaborate hexamers at the cell surface of confluent endothelial cells. Second, mutations were introduced within the extracellular part of VE-cadherin to destabilize the hexamer. Following an in vitro screening, three mutants were selected, among which, one is able to elaborate only dimers. The selected mutations were expressed as C-terminal green fluorescent protein fusions in CHO cells. Despite their capacity to elaborate nascent cell-cell contacts, the mutants seem to be rapidly degraded and/or internalized. Altogether, our results suggest that the formation of VE-cadherin hexamers protects this receptor and might allow the elaboration of mature endothelial cell-cell junctions

A novel family of transmembrane proteins interacting with beta subunits of the Na,K-ATPase

We characterized a family consisting of four mammalian proteins of unknown function (NKAIN1, 2, 3 and 4) and a single Drosophila ortholog dNKAIN. Aside from highly conserved transmembrane domains, NKAIN proteins contain no characterized functional domains. Striking amino acid conservation in the first two transmembrane domains suggests that these proteins are likely to function within the membrane bilayer. NKAIN family members are neuronally expressed in multiple regions of the mouse brain, although their expression is not ubiquitous. We demonstrate that mouse NKAIN1 interacts with the beta1 subunit of the Na,K-ATPase, whereas Drosophila ortholog dNKAIN interacts with Nrv2.2, a Drosophila homolog of the Na,K-ATPase beta subunits. We also show that NKAIN1 can form a complex with another beta subunit-binding protein, MONaKA, when binding to the beta1 subunit of the Na,K-ATPase. Our results suggest that a complex between mammalian NKAIN1 and MONaKA is required for NKAIN function, which is carried out by a single protein, dNKAIN, in Drosophila. This hypothesis is supported by the fact that dNKAIN, but not NKAIN1, induces voltage-independent amiloride-insensitive Na(+)-specific conductance that can be blocked by lanthanum. Drosophila mutants with decreased dNKAIN expression due to a P-element insertion in the dNKAIN gene exhibit temperature-sensitive paralysis, a phenotype also caused by mutations in the Na,K-ATPase alpha subunit and several ion channels. The neuronal expression of NKAIN proteins, their membrane localization and the temperature-sensitive paralysis of NKAIN Drosophila mutants strongly suggest that this novel protein family may be critical for neuronal function

IFN stimulated gene expression in the liver is a better predictor of treatment response in chronic hepatitis c than the IL28b (IFN lambda 3) genotype

Background: Therapy of chronic hepatitis C (CHC) with pegIFNa/ribavirin achieves sustained virologic response (SVR) in ~55%. Pre-activation of the endogenous interferon system in the liver is associated non-response (NR). Recently, genome-wide association studies described associations of allelic variants near the IL28B (IFNλ3) gene with treatment response and with spontaneous clearance of the virus. We investigated if the IL28B genotype determines the constitutive expression of IFN stimulated genes (ISGs) in the liver of patients with CHC. Methods: We genotyped 93 patients with CHC for 3 IL28B single nucleotide polymorphisms (SNPs, rs12979860, rs8099917, rs12980275), extracted RNA from their liver biopsies and quantified the expression of IL28B and of 8 previously identified classifier genes which discriminate between SVR and NR (IFI44L, RSAD2, ISG15, IFI22, LAMP3, OAS3, LGALS3BP and HTATIP2). Decision tree ensembles in the form of a random forest classifier were used to calculate the relative predictive power of these different variables in a multivariate analysis. Results: The minor IL28B allele (bad risk for treatment response) was significantly associated with increased expression of ISGs, and, unexpectedly, with decreased expression of IL28B. Stratification of the patients into SVR and NR revealed that ISG expression was conditionally independent from the IL28B genotype, i.e. there was an increased expression of ISGs in NR compared to SVR irrespective of the IL28B genotype. The random forest feature score (RFFS) identified IFI27 (RFFS = 2.93), RSAD2 (1.88) and HTATIP2 (1.50) expression and the HCV genotype (1.62) as the strongest predictors of treatment response. ROC curves of the IL28B SNPs showed an AUC of 0.66 with an error rate (ERR) of 0.38. A classifier with the 3 best classifying genes showed an excellent test performance with an AUC of 0.94 and ERR of 0.15. The addition of IL28B genotype information did not improve the predictive power of the 3-gene classifier. Conclusions: IL28B genotype and hepatic ISG expression are conditionally independent predictors of treatment response in CHC. There is no direct link between altered IFNλ3 expression and pre-activation of the endogenous system in the liver. Hepatic ISG expression is by far the better predictor for treatment response than IL28B genotype

Breaking the VE-cadherin bonds

Exchanges between the blood compartment and the surrounding tissues require a tight regulation by the endothelial barrier. Recent reports inferred that VE-cadherin, an endothelial specific cell-cell adhesion molecule, plays a pivotal role in the formation, maturation and remodeling of the vascular wall. Indeed, a growing number of permeability inducing factors (PIFs) was shown to elicit signaling mechanisms culminating in VE-cadherin destabilization and global alteration of the junctional architecture. Conversely, anti-PIFs protect from VE-cadherin disruption and enhance cell cohesion. These findings provide evidence on how endothelial cell-cell junctions impact the vascular network, and change our perception about normal and aberrant angiogenesis

Regulation of the Cardiac Na+/K+ ATPase by Phospholemman

Hansraj Dhayan, Rajender Kumar, Andreas Kukol, ‘Regulation of the Cardiac Na+/K+ ATPase by Phospholemman’, in Sajal Chakraborti, Naranjan Dhalla, eds., Regulation of Membrane Na+-K+ ATPase, (Switzerland: Springer, 2016), ISBN 978-3-319-24748-9, eISBN 978-3-319-24750-2.Peer reviewe

A genetic validation study reveals a role of vitamin D metabolism in the response to interferon-alfa-based therapy of chronic hepatitis C

Background: To perform a comprehensive study on the relationship between vitamin D metabolism and the response to interferon-α-based therapy of chronic hepatitis C. Methodology/Principal Findings: Associations between a functionally relevant polymorphism in the gene encoding the vitamin D 1α-hydroxylase (CYP27B1-1260 rs10877012) and the response to treatment with pegylated interferon-α (PEG-IFN-α) and ribavirin were determined in 701 patients with chronic hepatitis C. In addition, associations between serum concentrations of 25-hydroxyvitamin D3 (25[OH]D3) and treatment outcome were analysed. CYP27B1-1260 rs10877012 was found to be an independent predictor of sustained virologic response (SVR) in patients with poor-response IL28B genotypes (15% difference in SVR for rs10877012 genotype AA vs. CC, p = 0.02, OR = 1.52, 95% CI = 1.061–2.188), but not in patients with favourable IL28B genotype. Patients with chronic hepatitis C showed a high prevalence of vitamin D insufficiency (25[OH]D3<20 ng/mL) during all seasons, but 25(OH)D3 serum levels were not associated with treatment outcome. Conclusions/Significance: Our study suggests a role of bioactive vitamin D (1,25[OH]2D3, calcitriol) in the response to treatment of chronic hepatitis C. However, serum concentration of the calcitriol precursor 25(OH)D3 is not a suitable predictor of treatment outcome