Mutations in the nucleotide binding and hydrolysis domains of Helicobacter pylori MutS2 lead to altered biochemical activities and inactivation of its in vivo function

Background: Helicobacter pylori MutS2 (HpMutS2), an inhibitor of recombination during transformation is a non-specific nuclease with two catalytic sites, both of which are essential for its anti-recombinase activity. Although HpMutS2 belongs to a highly conserved family of ABC transporter ATPases, the role of its ATP binding and hydrolysis activities remains elusive. Results: To explore the putative role of ATP binding and hydrolysis activities of HpMutS2 we specifically generated point mutations in the nucleotide-binding Walker-A (HpMutS2-G338R) and hydrolysis Walker-B (HpMutS2-E413A) domains of the protein. Compared to wild-type protein, HpMutS2-G338R exhibited similar to 2.5-fold lower affinity for both ATP and ADP while ATP hydrolysis was reduced by similar to 3-fold. Nucleotide binding efficiencies of HpMutS2-E413A were not significantly altered; however the ATP hydrolysis was reduced by similar to 10-fold. Although mutations in the Walker-A and Walker-B motifs of HpMutS2 only partially reduced its ability to bind and hydrolyze ATP, we demonstrate that these mutants not only exhibited alterations in the conformation, DNA binding and nuclease activities of the protein but failed to complement the hyper-recombinant phenotype displayed by mutS2-disrupted strain of H. pylori. In addition, we show that the nucleotide cofactor modulates the conformation, DNA binding and nuclease activities of HpMutS2. Conclusions: These data describe a strong crosstalk between the ATPase, DNA binding, and nuclease activities of HpMutS2. Furthermore these data show that both, ATP binding and hydrolysis activities of HpMutS2 are essential for the in vivo anti-recombinase function of the protein.

New insight into the architecture of oxy-anion pocket in unliganded conformation of GAT domains: A MD-simulation study

Human Guanine Monophosphate Synthetase (hGMPS) converts XMP to GMP, and acts as a bifunctional enzyme with N-terminal ``glutaminase'' (GAT) and C-terminal ``synthetase'' domain. The enzyme is identified as a potential target for anticancer and immunosuppressive therapies. GAT domain of enzyme plays central role in metabolism, and contains conserved catalytic residues Cys104, His190, and Glu192. MD simulation studies on GAT domain suggest that position of oxyanion in unliganded conformation is occupied by one conserved water molecule (W1), which also stabilizes that pocket. This position is occupied by a negatively charged atom of the substrate or ligand in ligand bound crystal structures. In fact, MD simulation study of Ser75 to Val indicates that W1 conserved water molecule is stabilized by Ser75, while Thr152, and His190 also act as anchor residues to maintain appropriate architecture of oxyanion pocket through water mediated H-bond interactions. Possibly, four conserved water molecules stabilize oxyanion hole in unliganded state, but they vacate these positions when the enzyme (hGMPS)-substrate complex is formed. Thus this study not only reveals functionally important role of conserved water molecules in GAT domain, but also highlights essential role of other non-catalytic residues such as Ser75 and Thr152 in this enzymatic domain. The results from this computational study could be of interest to experimental community and provide a testable hypothesis for experimental validation. Conserved sites of water molecules near and at oxyanion hole highlight structural importance of water molecules and suggest a rethink of the conventional definition of chemical geometry of inhibitor binding site.

Homogeneous Hermitian holomorphic vector bundles and the Cowen-Douglas class over bounded symmetric domains

It is known that all the vector bundles of the title can be obtained by holomorphic induction from representations of a certain parabolic group on finite-dimensional inner product spaces. The representations, and the induced bundles, have composition series with irreducible factors. We write down an equivariant constant coefficient differential operator that intertwines the bundle with the direct sum of its irreducible factors. As an application, we show that in the case of the closed unit ball in C-n all homogeneous n-tuples of Cowen-Douglas operators are similar to direct sums of certain basic n-tuples. (c) 2015 Academie des sciences. Published by Elsevier Masson SAS. All rights reserved.

Domains beyond the grain boundary

Thin films of inorganic materials are used in diverse applications, typically in polycrystalline form due to their relatively simple production. We have used enhanced piezoresponse force microscopy to investigate the domain distribution within neighbouring grains in thin polycrystalline films of the ferroelectric-ferroelastic system lead zirconate titanate (PZT). We demonstrate that domains are organized into areas with a correlated alignment of the ferroelastic and ferroelectric domains, spanning multiple grain boundaries. We present five typical arrangements of such structures: azimuthal, radial, gradient, and short- and long- range linear domain organizations. Moreover, we discuss the mechanical and electrical constraints that dictate these structures. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

La política exterior de Raúl Alfonsín (1983- 1989) : un balance aproximativo

Este artículo tiene como objetivo realizar un balance de la política exterior de Raúl Alfonsín (1983-1989). Dicho análisis se sustentará en una contextualización histórica de su acción externa, en los fundamentos y creencias que sostuvieron su acción internacional y en sus objetivos principales. La reconstrucción en este punto de la investigación se basa especialmente en fuentes bibliográficas, dado que forma parte de los aspectos teóricos de un análisis más amplio, que incluye un análisis de casos de dicho período.

De Jiménez de Asúa a Perón : sus exilios como componentes de la política exterior hispano-argentina

Resumen: El compromiso vital de Jiménez de Asúa con sus ideas y con la España republicana, lanzará al eminente jurista al exilio en la Argentina, donde su prestigio intelectual le permitirá retomar su vida académica. Al encarnar la oposición legal en el exterior contra el solidificado régimen franquista, colocará su protagonismo en una situación cercana al poder desestabilizador del exilio en España del mismo Perón. La política exterior de la Argentina y de España, por diferentes vías de intervención, pero sin definiciones drásticas, procurará influir indirectamente para neutralizar a ambos.

Antiamericanismo y la política exterior estadounidense

Conferencia dada en la Pontificia Universidad Católica Argentina en el mes de junio de 2014: Existen quienes piensan que venir a la Argentina para dar una conferencia sobre el anti-americanismo es un poco como ir a Londres para dar una conferencia sobre el té. Los argentinos además de ser grandes candidatos para ganar la copa del mundo en fútbol, según se dice, son campeones en anti-norteamericanismo. Y es verdad que Argentina sigue siendo el país de América Latina con menor aprobación de los Estados Unidos. ¿Es esto evidencia del anti-americanismo en Argentina? Antes de contestar, debemos reconocer la dificultad que implica hablar del anti-americanismo en América Latina, ya que al identificar como “los americanos” a mis compatriotas, estaríamos apropiando simbólicamente a todo un continente. Ustedes también son americanos, y la monopolización del gentilicio por parte de una sola nación de las Américas refleja la lamentable actitud inconsciente de superioridad, que es una fuente de la falta de entendimiento entre norte y sur. Sin embargo, como es la palabra usada en mi país, voy a seguir usándola y problematizándola.

Rigorous finite-time guarantees for optimization on continuous domains by simulated annealing

Simulated annealing is a popular method for approaching the solution of a global optimization problem. Existing results on its performance apply to discrete combinatorial optimization where the optimization variables can assume only a finite set of possible values. We introduce a new general formulation of simulated annealing which allows one to guarantee finite-time performance in the optimization of functions of continuous variables. The results hold universally for any optimization problem on a bounded domain and establish a connection between simulated annealing and up-to-date theory of convergence of Markov chain Monte Carlo methods on continuous domains. This work is inspired by the concept of finite-time learning with known accuracy and confidence developed in statistical learning theory.

Repatriamento de capitais do exterior

Seminário, realizado em 5 de junho de 2012, sobre o Projeto de Lei nº 113/2003, que dispõe sobre o repatriamento de recursos depositados no exterior, e o Projeto de Lei nº 5.228/2005, que institui anistia fiscal sobre a legalização ou repatriamento de recursos mantidos no exterior não declarados e extingue a punibilidade dos delitos a eles relativos.

Análise da proposta de "política de desburocratização das exportações", apresentada pela Associação de Comércio Exterior do Brasil - AEB

Consultoria Legislativa - Área IX - Política e Planejamento Econômicos, Desenvolvimento Econômico, Economia Internacional.

Elaboração de projeto de lei concedendo cidadania brasileira para filhos e netos de brasileiros natos que nasceram no exterior, sob a forma de naturalização, a exemplo da lei italiana [lei do oriundo]

Consultoria Legislativa - Área XVIII - Direito Internacional Público, Relações Internacionais.

Brasileiros no exterior

Consultoria Legislativa - Área XVIII - Direito Internacional Público, Relações Internacionais.

Domain decomposition hybrid method for numerical simulation of bluff body flows:theoretical model and application

The discrete vortex method is not capable of precisely predicting the bluff body flow separation and the fine structure of flow field in the vicinity of the body surface. In order to make a theoretical improvement over the method and to reduce the difficulty in finite-difference solution of N-S equations at high Reynolds number, in the present paper, we suggest a new numerical simulation model and a theoretical method for domain decomposition hybrid combination of finite-difference method and vortex method. Specifically, the full flow. field is decomposed into two domains. In the region of O(R) near the body surface (R is the characteristic dimension of body), we use the finite-difference method to solve the N-S equations and in the exterior domain, we take the Lagrange-Euler vortex method. The connection and coupling conditions for flow in the two domains are established. The specific numerical scheme of this theoretical model is given. As a preliminary application, some numerical simulations for flows at Re=100 and Re-1000 about a circular cylinder are made, and compared with the finite-difference solution of N-S equations for full flow field and experimental results, and the stability of the solution against the change of the interface between the two domains is examined. The results show that the method of the present paper has the advantage of finite-difference solution for N-S equations in precisely predicting the fine structure of flow field, as well as the advantage of vortex method in efficiently computing the global characteristics of the separated flow. It saves computer time and reduces the amount of computation, as compared with pure N-S equation solution. The present method can be used for numerical simulation of bluff body flow at high Reynolds number and would exhibit even greater merit in that case.