Identification of allosteric hotspots regulating the ribosomal RNA binding by antibiotic resistance-conferring Erm methyltransferases

Antibiotic resistance via epigenetic methylation of ribosomal RNA is one of the most prevalent strategies adopted by multidrug resistant pathogens. The erythromycin-resistance methyltransferase (Erm) methylates rRNA at the conserved A2058 position and imparts resistance to macrolides such as erythromycin. However, the precise mechanism adopted by Erm methyltransferases for locating the target base within a complicated rRNA scaffold remains unclear. Here, we show that a conserved RNA architecture, including specific bulge sites, present more than 15 Å from the reaction center, is key to methylation at the pathogenic site. Using a set of RNA sequences site-specifically labeled by fluorescent nucleotide surrogates, we show that base flipping is a prerequisite for effective methylation and that distal bases assist in the recognition and flipping at the reaction center. The Erm–RNA complex model revealed that intrinsically flipped-out bases in the RNA serve as a putative anchor point for the Erm. Molecular dynamic simulation studies demonstrated the RNA undergoes a substantial change in conformation to facilitate an effective protein–rRNA handshake. This study highlights the importance of unique architectural features exploited by RNA to impart fidelity to RNA methyltransferases via enabling allosteric crosstalk. Moreover, the distal trigger sites identified here serve as attractive hotspots for the development of combination drug therapy aimed at reversing resistance

Nucleic Acids Res

The HIV-1 transactivator of transcription (Tat) protein is thought to stimulate reverse transcription (RTion). The Tat protein and, more specifically, its (44-61) domain were recently shown to promote the annealing of complementary DNA sequences representing the HIV-1 transactivation response element TAR, named dTAR and cTAR, that plays a key role in RTion. Moreover, the kinetic mechanism of the basic Tat(44-61) peptide in this annealing further revealed that this peptide constitutes a representative nucleic acid annealer. To further understand the structure-activity relationships of this highly conserved domain, we investigated by electrophoresis and fluorescence approaches the binding and annealing properties of various Tat(44-61) mutants. Our data showed that the Tyr47 and basic residues of the Tat(44-61) domain were instrumental for binding to cTAR through stacking and electrostatic interactions, respectively, and promoting its annealing with dTAR. Furthermore, the annealing efficiency of the mutants clearly correlates with their ability to rapidly associate and dissociate the complementary oligonucleotides and to promote RTion. Thus, transient and dynamic nucleic acid interactions likely constitute a key mechanistic component of annealers and the role of Tat in the late steps of RTion. Finally, our data suggest that Lys50 and Lys51 acetylation regulates Tat activity in RTion

Structural insights into the cTAR DNA recognition by the HIV-1 nucleocapsid protein: role of sugar deoxyriboses in the binding polarity of NC

An essential step of the reverse transcription of the HIV-1 genome is the first strand transfer that requires the annealing of the TAR RNA hairpin to the cTAR DNA hairpin. HIV-1 nucleocapsid protein (NC) plays a crucial role by facilitating annealing of the complementary hairpins. Using nuclear magnetic resonance and gel retardation assays, we investigated the interaction between NC and the top half of the cTAR DNA (mini-cTAR). We show that NC(11-55) binds the TGG sequence in the lower stem that is destabilized by the adjacent internal loop. The 5′ thymine interacts with residues of the N-terminal zinc knuckle and the 3′ guanine is inserted in the hydrophobic plateau of the C-terminal zinc knuckle. The TGG sequence is preferred relative to the apical and internal loops containing unpaired guanines. Investigation of the DNA–protein contacts shows the major role of hydrophobic interactions involving nucleobases and deoxyribose sugars. A similar network of hydrophobic contacts is observed in the published NC:DNA complexes, whereas NC contacts ribose differently in NC:RNA complexes. We propose that the binding polarity of NC is related to these contacts that could be responsible for the preferential binding to single-stranded nucleic acids

Multiple melting stages and refertilization as indicators for ridge to subduction formation: The New Caledonia ophiolite

International audienceThe origin of the New Caledonia ophiolite (South West Pacific), one of the largest in the world, is controversial. This nappe of ultramafic rocks (300 km long, 50 km wide and 2 km thick) is thrust upon a smaller nappe (Poya terrane) composed of basalts from mid-ocean ridges (MORB), back arc basins (BABB) and ocean islands (OIB). This nappe was tectonically accreted from the subducting plate prior and during the obduction of the ultramafic nappe. The bulk of the ophiolite is composed of highly depleted harzburgites (± dunites) with characteristic U-shaped bulk-rock rare-earth element (REE) patterns that are attributed to their formation in a forearc environment. In contrast, the origin of spoon-shaped REE patterns of lherzolites in the northernmost klippes was unclear. Our new major element and REE data on whole rocks, spinel and clinopyroxene establish the abyssal affinity of these lherzolites. Significant LREE enrichment in the lherzolites is best explained by partial melting in a spreading ridge, followed by near in-situ refertilization from deeper mantle melts. Using equilibrium melting equations, we show that melts extracted from these lherzolites are compositionally similar to the MORB of the Poya terrane. This is used to infer that the ultramafic nappe and the mafic Poya terrane represent oceanic lithosphere of a single marginal basin that formed during the late Cretaceous. In contrast, our spinel data highlights the strong forearc affinities of the most depleted harzburgites whose compositions are best modeled by hydrous melting of a source that had previously experienced depletion in a spreading ridge. The New Caledonian boninites probably formed during this second stage of partial melting. The two melting events in the New Caledonia ophiolite record the rapid transition from oceanic accretion to convergence in the South Loyalty Basin during the Late Paleocene, with initiation of a new subduction zone at or near the ridge axis

Coronary microvascular resistance: methods for its quantification in humans

Coronary microvascular dysfunction is a topic that has recently gained considerable interest in the medical community owing to the growing awareness that microvascular dysfunction occurs in a number of myocardial disease states and has important prognostic implications. With this growing awareness, comes the desire to accurately assess the functional capacity of the coronary microcirculation for diagnostic purposes as well as to monitor the effects of therapeutic interventions that are targeted at reversing the extent of coronary microvascular dysfunction. Measurements of coronary microvascular resistance play a pivotal role in achieving that goal and several invasive and noninvasive methods have been developed for its quantification. This review is intended to provide an update pertaining to the methodology of these different imaging techniques, including the discussion of their strengths and weaknesses

GRIP. Europe des armes. Trafics et exportations vers le Tiers Monde

Boudier Christian. GRIP. Europe des armes. Trafics et exportations vers le Tiers Monde . In: Politique étrangère, n°3 - 1989 - 54ᵉannée. pp. 518-519

Amérique latine

Boudier Christian. Amérique latine. In: Politique étrangère, n°1 - 1988 - 53ᵉannée. pp. 295-296

Amérique latine

Boudier Christian. Amérique latine. In: Politique étrangère, n°1 - 1988 - 53ᵉannée. pp. 295-296

Denis Rolland. Vichy et la France libre au Mexique : guerre, cultures et propagande pendant la Seconde Guerre mondiale

Boudier Christian. Denis Rolland. Vichy et la France libre au Mexique : guerre, cultures et propagande pendant la Seconde Guerre mondiale. In: Politique étrangère, n°3 - 1990 - 55ᵉannée. pp. 717-718

Cartes

Boudier Christian. Cartes. In: Politique étrangère, n°2 - 1987 - 52ᵉannée. pp. 402-403