How mutations in tRNA distant from the anticodon affect the fidelity of decoding

The ribosome converts genetic information into protein by selecting aminoacyl tRNAs whose anticodons base-pair to an mRNA codon. Mutations in the tRNA body can perturb this process and affect fidelity. The Hirsh suppressor is a well-studied tRNA^(Trp) harboring a G24A mutation that allows readthrough of UGA stop codons. Here we present crystal structures of the 70S ribosome complexed with EF-Tu and aminoacyl tRNA (native tRNA^(Trp), G24A tRNA^(Trp) or the miscoding A9C tRNA^(Trp)) bound to cognate UGG or near-cognate UGA codons, determined at 3.2-Å resolution. The A9C and G24A mutations lead to miscoding by facilitating the distortion of tRNA required for decoding. A9C accomplishes this by increasing tRNA flexibility, whereas G24A allows the formation of an additional hydrogen bond that stabilizes the distortion. Our results also suggest that each native tRNA will adopt a unique conformation when delivered to the ribosome that allows accurate decoding

The isolation of novel "Erwinia" phages and their use in the study of bacterial phytopathogenicity

A number of bacteriophages were isolated on the "soft rot" phytopathogens Erwinia carotovora subsp. atroseptica SCRI1043 and Erwinia carotovora subsp. carotovora SCRI193. Several of these phages were used to obtain phage resistant mutants of SCRI1043, in order to investigate the role of the bacterial cell surface in virulence. While a number of phenotypic properties relating to pathogenicity and virulence of this strain have already been uncovered, little is known about the role of the cell surface in virulence. It was hoped that the use of phages would allow selection of mutants altered in both cell surface and virulence. Two phage resistant mutants, A5/22 and A5/8, exhibited reduced virulence when inoculated into potato plants, and were investigated further. Both mutants showed pleiotropic phenotypes. As well as reduced virulence and phage resistance, these mutants showed a number of other phenotypic alterations including, a reduction in the production of plant cell wall degrading enzymes, increased sensitivity to surface active agents, alterations in lipopolysaccharide and outer membrane protein profiles and reduced motility. A5/22 also exhibited bacteriostasis in the presence of galactose. Mutant A5/22 was more severely affected in its virulence than A5/8, which reflected in its greater deviation from the wild type phenotype. While no one phenotypic alteration could be directly associated with the reduced virulence of either mutant, a combination of several phenotypes may have been responsible. The phages isolated in this study were the first reported for these strains of Erwinia, and were therefore characterised under a number of criteria. All phages were grouped on the basis of structural morphology, restriction endonuclease digestion and host range. This is the first detailed characterisation of phages for Erwinia carotovora subsp. atroseptica. All isolated phages were tested for generalised transduction, a method of molecular genetic analysis so far unavailable to Erwinia carotovora subsp. atroseptica SCRI1043 and Erwinia carotovora subsp. carotovora SCRI193. Two phages, ØKP and ØMl, were capable of generalised transduction in SCRI193 and SCRI1043 respectively. Both these phages were characterised and transducing frequencies improved. ØMl is the first transducing phage reported for Erwinia carotovora subsp. atroseptica and ØKP is only the second for Erwinia carotovora subsp. carotovora. Both phages are now being used extensively in the laboratory

The Crystal Structure of the Ribosome Bound to EF-Tu and Aminoacyl-tRNA

The ribosome selects a correct transfer RNA (tRNA) for each amino acid added to the polypeptide chain, as directed by messenger RNA. Aminoacyl-tRNA is delivered to the ribosome by elongation factor Tu (EF-Tu), which hydrolyzes guanosine triphosphate (GTP) and releases tRNA in response to codon recognition. The signaling pathway that leads to GTP hydrolysis upon codon recognition is critical to accurate decoding. Here we present the crystal structure of the ribosome complexed with EF-Tu and aminoacyl-tRNA, refined to 3.6 angstrom resolution. The structure reveals details of the tRNA distortion that allows aminoacyl-tRNA to interact simultaneously with the decoding center of the 30S subunit and EF-Tu at the factor binding site. A series of conformational changes in EF-Tu and aminoacyl-tRNA suggests a communication pathway between the decoding center and the guanosine triphosphatase center of EF-Tu

The Crystal Structure of the Ribosome Bound to EF-Tu and Aminoacyl-tRNA

The ribosome selects a correct transfer RNA (tRNA) for each amino acid added to the polypeptide chain, as directed by messenger RNA. Aminoacyl-tRNA is delivered to the ribosome by elongation factor Tu (EF-Tu), which hydrolyzes guanosine triphosphate (GTP) and releases tRNA in response to codon recognition. The signaling pathway that leads to GTP hydrolysis upon codon recognition is critical to accurate decoding. Here we present the crystal structure of the ribosome complexed with EF-Tu and aminoacyl-tRNA, refined to 3.6 angstrom resolution. The structure reveals details of the tRNA distortion that allows aminoacyl-tRNA to interact simultaneously with the decoding center of the 30S subunit and EF-Tu at the factor binding site. A series of conformational changes in EF-Tu and aminoacyl-tRNA suggests a communication pathway between the decoding center and the guanosine triphosphatase center of EF-Tu

Structural Diversity in Bacterial Ribosomes: Mycobacterial 70S Ribosome Structure Reveals Novel Features

Here we present analysis of a 3D cryo-EM map of the 70S ribosome from Mycobacterium smegmatis, a saprophytic cousin of the etiological agent of tuberculosis in humans, Mycobacterium tuberculosis. In comparison with the 3D structures of other prokaryotic ribosomes, the density map of the M. smegmatis 70S ribosome reveals unique structural features and their relative orientations in the ribosome. Dramatic changes in the periphery due to additional rRNA segments and extra domains of some of the peripheral ribosomal proteins like S3, S5, S16, L17, L25, are evident. One of the most notable features appears in the large subunit near L1 stalk as a long helical structure next to helix 54 of the 23S rRNA. The sharp upper end of this structure is located in the vicinity of the mRNA exit channel. Although the M. smegmatis 70S ribosome possesses conserved core structure of bacterial ribosome, the new structural features, unveiled in this study, demonstrates diversity in the 3D architecture of bacterial ribosomes. We postulate that the prominent helical structure related to the 23S rRNA actively participates in the mechanisms of translation in mycobacteria

Light isovector resonances in π-p →π-π-π+p at 190 GeV/c

We have performed the most comprehensive resonance-model fit of π-π-π+ states using the results of our previously published partial-wave analysis (PWA) of a large data set of diffractive-dissociation events from the reaction π-+p→π-π-π++precoil with a 190 GeV/c pion beam. The PWA results, which were obtained in 100 bins of three-pion mass, 0.5<2.5 GeV/c2, and simultaneously in 11 bins of the reduced four-momentum transfer squared, 0.1<1.0 (GeV/c)2, are subjected to a resonance-model fit using Breit-Wigner amplitudes to simultaneously describe a subset of 14 selected waves using 11 isovector light-meson states with JPC=0-+, 1++, 2++, 2-+, 4++, and spin-exotic 1-+ quantum numbers. The model contains the well-known resonances π(1800), a1(1260), a2(1320), π2(1670), π2(1880), and a4(2040). In addition, it includes the disputed π1(1600), the excited states a1(1640), a2(1700), and π2(2005), as well as the resonancelike a1(1420). We measure the resonance parameters mass and width of these objects by combining the information from the PWA results obtained in the 11 t′ bins. We extract the relative branching fractions of the ρ(770)π and f2(1270)π decays of a2(1320) and a4(2040), where the former one is measured for the first time. In a novel approach, we extract the t′ dependence of the intensity of the resonances and of their phases. The t′ dependence of the intensities of most resonances differs distinctly from the t′ dependence of the nonresonant components. For the first time, we determine the t′ dependence of the phases of the production amplitudes and confirm that the production mechanism of the Pomeron exchange is common to all resonances. We have performed extensive systematic studies on the model dependence and correlations of the measured physical parameters

Double J/ψJ/\psi production in pion-nucleon scattering at COMPASS

We present the study of the production of double $J/\psi$ mesons using COMPASS data collected with a 190 GeV/$c$ $\pi^-$ beam scattering off NH$_{3}$, Al and W targets. Kinematic distributions of the collected double $J/\psi$ events are analysed, and the double $J/\psi$ production cross section is estimated for each of the COMPASS targets. The results are compared to predictions from single- and double-parton scattering models as well as the pion intrinsic charm and the tetraquark exotic resonance hypotheses. It is demonstrated that the single parton scattering production mechanism gives the dominant contribution that is sufficient to describe the data. An upper limit on the double intrinsic charm content of pion is evaluated. No significant signatures that could be associated with exotic tetraquarks are found in the double $J/\psi$ mass spectrum.Comment: 12 pages, 4 figure

Searches for lepton-flavour-violating decays of the Higgs boson into eτ and μτ in \sqrt{s} = 13 TeV pp collisions with the ATLAS detector

Abstract This paper presents direct searches for lepton flavour violation in Higgs boson decays, H → eτ and H → μτ, performed using data collected with the ATLAS detector at the LHC. The searches are based on a data sample of proton-proton collisions at a centre-of-mass energy s $$\sqrt{s}$$ = 13 TeV, corresponding to an integrated luminosity of 138 fb−1. Leptonic (τ → ℓνℓντ) and hadronic (τ → hadrons ντ) decays of the τ-lepton are considered. Two background estimation techniques are employed: the MC-template method, based on data-corrected simulation samples, and the Symmetry method, based on exploiting the symmetry between electrons and muons in the Standard Model backgrounds. No significant excess of events is observed and the results are interpreted as upper limits on lepton-flavour-violating branching ratios of the Higgs boson. The observed (expected) upper limits set on the branching ratios at 95% confidence level, B $$\mathcal{B}$$ (H → eτ) < 0.20% (0.12%) and B $$\mathcal{B}$$ (H → μτ ) < 0.18% (0.09%), are obtained with the MC-template method from a simultaneous measurement of potential H → eτ and H → μτ signals. The best-fit branching ratio difference, B $$\mathcal{B}$$ (H → μτ) → B $$\mathcal{B}$$ (H → eτ), measured with the Symmetry method in the channel where the τ-lepton decays to leptons, is (0.25 ± 0.10)%, compatible with a value of zero within 2.5σ

Comparison of inclusive and photon-tagged jet suppression in 5.02 TeV Pb+Pb collisions with ATLAS

Parton energy loss in the quark–gluon plasma (QGP) is studied with a measurement of photon-tagged jet production in 1.7 nb−1 of Pb+Pb data and 260 pb−1 of pp data, both at sNN=5.02 TeV, with the ATLAS detector. The process pp →γ+jet+X and its analogue in Pb+Pb collisions is measured in events containing an isolated photon with transverse momentum (pT) above 50 GeV and reported as a function of jet pT. This selection results in a sample of jets with a steeply falling pT distribution that are mostly initiated by the showering of quarks. The pp and Pb+Pb measurements are used to report the nuclear modification factor, RAA, and the fractional energy loss, Sloss, for photon-tagged jets. In addition, the results are compared with the analogous ones for inclusive jets, which have a significantly smaller quark-initiated fraction. The RAA and Sloss values are found to be significantly different between those for photon-tagged jets and inclusive jets, demonstrating that energy loss in the QGP is sensitive to the colour-charge of the initiating parton. The results are also compared with a variety of theoretical models of colour-charge-dependent energy loss