The Bacillus anthracis arylamine N-acetyltransferase ((BACAN)NAT1) that inactivates sulfamethoxazole, reveals unusual structural features compared with the other NAT isoenzymes

AbstractArylamine N-acetyltransferases (NATs) are xenobiotic-metabolizing enzymes that biotransform arylamine drugs. The Bacillus anthracis (BACAN)NAT1 enzyme affords increased resistance to the antibiotic sulfamethoxazole through its acetylation. We report the structure of (BACAN)NAT1. Unexpectedly, endogenous coenzymeA was present in the active site. The structure suggests that, contrary to the other prokaryotic NATs, (BACAN)NAT1 possesses a 14-residue insertion equivalent to the “mammalian insertion”, a structural feature considered unique to mammalian NATs. Moreover, (BACAN)NAT1 structure shows marked differences in the mode of binding and location of coenzymeA when compared to the other NATs. This suggests that the mechanisms of cofactor recognition by NATs is more diverse than expected and supports the cofactor-binding site as being a unique subsite to target in drug design against bacterial NATs

The crystal structure of Trz1, the long form RNase Z from yeast.

tRNAs are synthesized as precursor RNAs that have to undergo processing steps to become functional. Yeast Trz1 is a key endoribonuclease involved in the 3΄ maturation of tRNAs in all domains of life. It is a member of the β-lactamase family of RNases, characterized by an HxHxDH sequence motif involved in coordination of catalytic Zn-ions. The RNase Z family consists of two subfamilies: the short (250-400 residues) and the long forms (about double in size). Short form RNase Z enzymes act as homodimers: one subunit embraces tRNA with a protruding arm, while the other provides the catalytic site. The long form is thought to contain two fused β-lactamase domains within a single polypeptide. Only structures of short form RNase Z enzymes are known. Here we present the 3.1 Å crystal structure of the long-form Trz1 from Saccharomyces cerevisiae. Trz1 is organized into two β-lactamase domains connected by a long linker. The N-terminal domain has lost its catalytic residues, but retains the long flexible arm that is important for tRNA binding, while it is the other way around in the C-terminal domain. Trz1 likely evolved from a duplication and fusion of the gene encoding the monomeric short form RNase Z

Specific GFP-binding artificial proteins ( Rep): a new tool for in vitro to live cell applications

International audienceA family of artificial proteins, named αRep, based on a natural family of helical repeat was previously designed. αRep members are efficiently expressed, folded and extremely stable proteins. A large αRep library was constructed creating proteins with a randomized interaction surface. In the present study, we show that the αRep library is an efficient source of tailor-made specific proteins with direct applications in biochemistry and cell biology. From this library, we selected by phage display αRep binders with nanomolar dissociation constants against the GFP. The structures of two independent αRep binders in complex with the GFP target were solved by X-ray crystallography revealing two totally different binding modes. The affinity of the selected αReps for GFP proved sufficient for practically useful applications such as pull-down experiments. αReps are disulfide free proteins and are efficiently and functionally expressed in eukaryotic cells: GFP-specific αReps are clearly sequestrated by their cognate target protein addressed to various cell compartments. These results suggest that αRep proteins with tailor-made specificity can be selected and used in living cells to track, modulate or interfere with intracellular processes

Aragonite saturation state in a continental shelf (Gulf of Cádiz, SW IberianPeninsula): Evidences of acidification in the coastal area

The spatiotemporal variability of aragonite saturation state (ΩAr) has been studied in the eastern shelf of the Gulf of Cádiz (GoC) (SW Iberian Peninsula). The study was carried out during the years 2014 and 2016 aboard twelve oceanographic cruises, along three or five transects, located between Cape Trafalgar and the Guadiana River. The GoC exhibited oversaturated of calcium carbonate with ΩAr mean values of 2.68 ± 0.30 in surface and 2.05 ± 0.15 in deep waters. pH, total alkalinity (TA), calcium concentration (Ca2+) and ΩAr showed a high variability within the surface mixed layer (SML, z 100 m), TA and Ca2+ concentration presented a conservative behaviour related to the distribution of the different water masses located in the GoC. The vertical variation of ΩAr also depends on the degree of mineralization of these water masses, obtaining the maximum values in the Subtropical North Atlantic Central Water (100–200 m), minimum values in the Subpolar North Atlantic Central Water (about 400 m), and intermediate values associated to the presence of the Mediterranean Water (>500 m). Results showed a significative acidification of the coastal areas, for those depths lower than 100 m from 2006 to 2016, with a mean decrease of pH and ΩAr of −0.0089 and −0.0552 yr−1, respectively. © 2021 The AuthorsThis work was funded by the Spanish CICYT (Spanish Program for Science and Technology) under the contract RTI2018-100865-B-C21 . Dolores Jiménez-López was financed by the University of Cádiz with a FPI fellowship (FPI-UCA) and Ana Sierra was financed by the Spanish Ministry of Education with a FPU fellowship (FPU2014-04048)

A survey of green plant tRNA 3'-end processing enzyme tRNase Zs, homologs of the candidate prostate cancer susceptibility protein ELAC2

<p>Abstract</p> <p>Background</p> <p>tRNase Z removes the 3'-trailer sequences from precursor tRNAs, which is an essential step preceding the addition of the CCA sequence. tRNase Z exists in the short (tRNase Z^S) and long (tRNase Z^L) forms. Based on the sequence characteristics, they can be divided into two major types: bacterial-type tRNase Z^Sand eukaryotic-type tRNase Z^L, and one minor type, <it>Thermotoga maritima </it>(TM)-type tRNase Z^S. The number of tRNase Zs is highly variable, with the largest number being identified experimentally in the flowering plant <it>Arabidopsis thaliana</it>. It is unknown whether multiple tRNase Zs found in <it>A. thaliana </it>is common to the plant kingdom. Also unknown is the extent of sequence and structural conservation among tRNase Zs from the plant kingdom.</p> <p>Results</p> <p>We report the identification and analysis of candidate tRNase Zs in 27 fully sequenced genomes of green plants, the great majority of which are flowering plants. It appears that green plants contain multiple distinct tRNase Zs predicted to reside in different subcellular compartments. Furthermore, while the bacterial-type tRNase Z^Ss are present only in basal land plants and green algae, the TM-type tRNase Z^Ss are widespread in green plants. The protein sequences of the TM-type tRNase Z^Ss identified in green plants are similar to those of the bacterial-type tRNase Z^Ss but have distinct features, including the TM-type flexible arm, the variant catalytic HEAT and HST motifs, and a lack of the PxKxRN motif involved in CCA anti-determination (inhibition of tRNase Z activity by CCA), which prevents tRNase Z cleavage of mature tRNAs. Examination of flowering plant chloroplast tRNA genes reveals that many of these genes encode partial CCA sequences. Based on our results and previous studies, we predict that the plant TM-type tRNase Z^Ss may not recognize the CCA sequence as an anti-determinant.</p> <p>Conclusions</p> <p>Our findings substantially expand the current repertoire of the TM-type tRNase Z^Ss and hint at the possibility that these proteins may have been selected for their ability to process chloroplast pre-tRNAs with whole or partial CCA sequences. Our results also support the coevolution of tRNase Zs and tRNA 3'-trailer sequences in plants.</p

Model-independent search for CP violation in D0→K−K+π−π+ and D0→π−π+π+π− decays

A search for CP violation in the phase-space structures of D0 and View the MathML source decays to the final states K−K+π−π+ and π−π+π+π− is presented. The search is carried out with a data set corresponding to an integrated luminosity of 1.0 fb−1 collected in 2011 by the LHCb experiment in pp collisions at a centre-of-mass energy of 7 TeV. For the K−K+π−π+ final state, the four-body phase space is divided into 32 bins, each bin with approximately 1800 decays. The p-value under the hypothesis of no CP violation is 9.1%, and in no bin is a CP asymmetry greater than 6.5% observed. The phase space of the π−π+π+π− final state is partitioned into 128 bins, each bin with approximately 2500 decays. The p-value under the hypothesis of no CP violation is 41%, and in no bin is a CP asymmetry greater than 5.5% observed. All results are consistent with the hypothesis of no CP violation at the current sensitivity

Search for the lepton-flavor-violating decays Bs0→e±μ∓ and B0→e±μ∓

A search for the lepton-flavor-violating decays Bs0→e±μ∓ and B0→e±μ∓ is performed with a data sample, corresponding to an integrated luminosity of 1.0  fb-1 of pp collisions at √s=7  TeV, collected by the LHCb experiment. The observed number of Bs0→e±μ∓ and B0→e±μ∓ candidates is consistent with background expectations. Upper limits on the branching fractions of both decays are determined to be B(Bs0→e±μ∓)101  TeV/c2 and MLQ(B0→e±μ∓)>126  TeV/c2 at 95% C.L., and are a factor of 2 higher than the previous bounds

Observation of the decay B+c→Bºsπ+

The result of a search for the decay B+c→Bºsπ+ is presented, using the Bºs→Ds-π+ and Bºs→J/ψϕ channels. The analysis is based on a data sample of pp collisions collected with the LHCb detector, corresponding to an integrated luminosity of 1  fb-1 taken at a center-of-mass energy of 7 TeV, and 2  fb-1 taken at 8 TeV. The decay B+c→Bºsπ+ is observed with significance in excess of 5 standard deviations independently in both decay channels. The measured product of the ratio of cross sections and branching fraction is [σ(Bc+)/σ(Bºs)]×B(Bc+→Bºsπ+)=[2.37±0.31 (stat)±0.11 (syst)-0.13+0.17(τBc+)]×10-3, in the pseudorapidity range 2<η(B)<5, where the first uncertainty is statistical, the second is systematic, and the third is due to the uncertainty on the Bc+ lifetime. This is the first observation of a B meson decaying to another B meson via the weak interaction

Study of B0(s)→K0Sh+h′− decays with first observation of B0s→K0SK±π∓ and B0s→K0Sπ+π−

A search for charmless three-body decays of B 0 and B0s mesons with a K0S meson in the final state is performed using the pp collision data, corresponding to an integrated luminosity of 1.0 fb−1, collected at a centre-of-mass energy of 7 TeV recorded by the LHCb experiment. Branching fractions of the B0(s)→K0Sh+h′− decay modes (h (′) = π, K), relative to the well measured B0→K0Sπ+π− decay, are obtained. First observation of the decay modes B0s→K0SK±π∓ and B0s→K0Sπ+π− and confirmation of the decay B0→K0SK±π∓ are reported. The following relative branching fraction measurements or limits are obtained B(B0→K0SK±π∓)B(B0→K0Sπ+π−)=0.128±0.017(stat.)±0.009(syst.), B(B0→K0SK+K−)B(B0→K0Sπ+π−)=0.385±0.031(stat.)±0.023(syst.), B(B0s→K0Sπ+π−)B(B0→K0Sπ+π−)=0.29±0.06(stat.)±0.03(syst.)±0.02(fs/fd), B(B0s→K0SK±π∓)B(B0→K0Sπ+π−)=1.48±0.12(stat.)±0.08(syst.)±0.12(fs/fd)B(B0s→K0SK+K−)B(B0→K0Sπ+π−)∈[0.004;0.068]at90%CL