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Native porphyran is a hybrid of porphryan and agarose. As a common element of edible seaweed, this algal galactan is a frequent component of the human diet. Bacterial members of the human gut microbiota have acquired polysaccharide utilization loci (PULs) that enable the metabolism of porphyran or agarose. However, the molecular mechanisms that underlie the deconstruction and use of native porphyran remains incompletely defined. Here, we have studied two human gut bacteria, porphyranolytic Bacteroides plebeius and agarolytic Bacteroides uniformis, that target native porphyran. This reveals an exo-based cycle of porphyran depolymerization that incorporates a keystone sulfatase. In both PULs this cycle also works together with a PUL-encoded agarose depolymerizing machinery to synergistically reduce native porphyran to monosaccharides. This provides a framework for understanding the deconstruction of a hybrid algal galactan, and insight into the competitive and/or syntrophic relationship of gut microbiota members that target rare nutrients.
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Microbioma Gastrointestinal , Bactérias/metabolismo , Galactanos , Humanos , Polissacarídeos/metabolismo , SefaroseRESUMO
Glycosidic linkage analysis was conducted on the unfractionated polysaccharides in alcohol-insoluble residues (AIRs) prepared from six red seaweeds (Gracilariopsis sp., Prionitis sp., Mastocarpus papillatus, Callophyllis sp., Mazzaella splendens, and Palmaria palmata) using GC-MS/FID analysis of partially methylated alditol acetates (PMAAs). The cell walls of P. palmata primarily contained mixed-linkage xylans and small amounts of sulfated galactans and cellulose. In contrast, the unfractionated polysaccharides of the other five species were rich in galactans displaying diverse 3,6-anhydro-galactose and galactose linkages with varied sulfation patterns. Different levels of cellulose were also observed. This glycosidic linkage method offers advantages for cellulose analysis over traditional monosaccharide analysis that is known for underrepresenting glucose in crystalline cellulose. Relative linkage compositions calculated from GC-MS and GC-FID measurements showed that anhydro sugar linkages generated more responses in the latter detection method. This improved linkage workflow presents a useful tool for studying polysaccharide structural variations across red seaweed species. Furthermore, for the first time, relative linkage compositions from GC-MS and GC-FID measurements, along with normalized FID and total ion current (TIC) chromatograms without peak assignments, were analyzed using principal component analysis (PCA) as a proof-of-concept demonstration of the technique's potential to differentiate various red seaweed species.
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Cromatografia Gasosa-Espectrometria de Massas , Polissacarídeos , Rodófitas , Alga Marinha , Polissacarídeos/química , Alga Marinha/química , Cromatografia Gasosa-Espectrometria de Massas/métodos , Rodófitas/química , Metilação , Glicosídeos/químicaRESUMO
Gardnerella spp. are associated with bacterial vaginosis in which normally dominant lactobacilli are replaced with facultative and anaerobic bacteria, including Gardnerella spp. Co-occurrence of multiple species of Gardnerella is common in the vagina, and competition for nutrients such as glycogen likely contributes to the differential abundances of Gardnerella spp. Glycogen must be digested into smaller components for uptake, a process that depends on the combined action of glycogen-degrading enzymes. In this study, the ability of culture supernatants of 15 isolates of Gardnerella spp. to produce glucose, maltose, maltotriose, and maltotetraose from glycogen was demonstrated. Carbohydrate-active enzymes (CAZymes) were identified bioinformatically in Gardnerella proteomes using dbCAN2. Identified proteins included a single-domain α-amylase (EC 3.2.1.1) (encoded by all 15 isolates) and an α-amylase-pullulanase (EC 3.2.1.41) containing amylase, carbohydrate binding modules, and pullulanase domains (14/15 isolates). To verify the sequence-based functional predictions, the amylase and pullulanase domains of the α-amylase-pullulanase and the single-domain α-amylase were each produced in Escherichia coli. The α-amylase domain from the α-amylase-pullulanase released maltose, maltotriose, and maltotetraose from glycogen, and the pullulanase domain released maltotriose from pullulan and maltose from glycogen, demonstrating that the Gardnerella α-amylase-pullulanase is capable of hydrolyzing α-1,4 and α-1,6 glycosidic bonds. Similarly, the single-domain α-amylase protein also produced maltose, maltotriose, and maltotetraose from glycogen. Our findings show that Gardnerella spp. produce extracellular amylase enzymes as "public goods" that can digest glycogen into maltose, maltotriose, and maltotetraose that can be used by the vaginal microbiota. IMPORTANCE Increased abundance of Gardnerella spp. is a diagnostic characteristic of bacterial vaginosis, an imbalance in the human vaginal microbiome associated with troubling symptoms, and negative reproductive health outcomes, including increased transmission of sexually transmitted infections and preterm birth. Competition for nutrients is likely an important factor in causing dramatic shifts in the vaginal microbial community, but little is known about the contribution of bacterial enzymes to the metabolism of glycogen, a major food source available to vaginal bacteria. The significance of our research is characterizing the activity of enzymes conserved in Gardnerella species that contribute to the ability of these bacteria to utilize glycogen.
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Microbiota , Nascimento Prematuro , Vaginose Bacteriana , Feminino , Humanos , alfa-Amilases/metabolismo , Bactérias/metabolismo , Domínio Catalítico , Gardnerella , Glicogênio/metabolismo , Maltose , Vagina/microbiologia , Vaginose Bacteriana/microbiologiaRESUMO
This Letter reports the observation of τ-lepton-pair production in ultraperipheral lead-lead collisions Pb+PbâPb(γγâττ)Pb and constraints on the τ-lepton anomalous magnetic moment a_{τ}. The dataset corresponds to an integrated luminosity of 1.44 nb^{-1} of LHC Pb+Pb collisions at sqrt[s_{NN}]=5.02 TeV recorded by the ATLAS experiment in 2018. Selected events contain one muon from a τ-lepton decay, an electron or charged-particle track(s) from the other τ-lepton decay, little additional central-detector activity, and no forward neutrons. The γγâττ process is observed in Pb+Pb collisions with a significance exceeding 5 standard deviations and a signal strength of µ_{ττ}=1.03_{-0.05}^{+0.06} assuming the standard model value for a_{τ}. To measure a_{τ}, a template fit to the muon transverse-momentum distribution from τ-lepton candidates is performed, using a dimuon (γγ⵵) control sample to constrain systematic uncertainties. The observed 95% confidence-level interval for a_{τ} is -0.057
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Jet quenching is the process of color-charged partons losing energy via interactions with quark-gluon plasma droplets created in heavy-ion collisions. The collective expansion of such droplets is well described by viscous hydrodynamics. Similar evidence of collectivity is consistently observed in smaller collision systems, including pp and p+Pb collisions. In contrast, while jet quenching is observed in Pb+Pb collisions, no evidence has been found in these small systems to date, raising fundamental questions about the nature of the system created in these collisions. The ATLAS experiment at the Large Hadron Collider has measured the yield of charged hadrons correlated with reconstructed jets in 0.36 nb^{-1} of p+Pb and 3.6 pb^{-1} of pp collisions at 5.02 TeV. The yields of charged hadrons with p_{T}^{ch}>0.5 GeV near and opposite in azimuth to jets with p_{T}^{jet}>30 or 60 GeV, and the ratios of these yields between p+Pb and pp collisions, I_{pPb}, are reported. The collision centrality of p+Pb events is categorized by the energy deposited by forward neutrons from the struck nucleus. The I_{pPb} values are consistent with unity within a few percent for hadrons with p_{T}^{ch}>4 GeV at all centralities. These data provide new, strong constraints that preclude almost any parton energy loss in central p+Pb collisions.
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This Letter reports the observation of single top quarks produced together with a photon, which directly probes the electroweak coupling of the top quark. The analysis uses 139 fb^{-1} of 13 TeV proton-proton collision data collected with the ATLAS detector at the Large Hadron Collider. Requiring a photon with transverse momentum larger than 20 GeV and within the detector acceptance, the fiducial cross section is measured to be 688±23(stat) _{-71}^{+75}(syst) fb, to be compared with the standard model prediction of 515_{-42}^{+36} fb at next-to-leading order in QCD.
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A search for a long-lived, heavy neutral lepton (N) in 139 fb^{-1} of sqrt[s]=13 TeV pp collision data collected by the ATLAS detector at the Large Hadron Collider is reported. The N is produced via WâNµ or WâNe and decays into two charged leptons and a neutrino, forming a displaced vertex. The N mass is used to discriminate between signal and background. No signal is observed, and limits are set on the squared mixing parameters of the N with the left-handed neutrino states for the N mass range 3 GeV
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A test of CP invariance in Higgs boson production via vector-boson fusion has been performed in the Hâγγ channel using 139 fb^{-1} of proton-proton collision data at sqrt[s]=13 TeV collected by the ATLAS detector at the LHC. The optimal observable method is used to probe the CP structure of interactions between the Higgs boson and electroweak gauge bosons, as described by an effective field theory. No sign of CP violation is observed in the data. Constraints are set on the parameters describing the strength of the CP-odd component in the coupling between the Higgs boson and the electroweak gauge bosons in two effective field theory bases: d[over Ë] in the HISZ basis and c_{HW[over Ë]} in the Warsaw basis. The results presented are the most stringent constraints on CP violation in the coupling between Higgs and weak bosons. The 95% C.L. constraint on d[over Ë] is derived for the first time and the 95% C.L. constraint on c_{HW[over Ë]} has been improved by a factor of 5 compared to the previous measurement.
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The metabolism of carbohydrate polymers drives microbial diversity in the human gut microbiota. It is unclear, however, whether bacterial consortia or single organisms are required to depolymerize highly complex glycans. Here we show that the gut bacterium Bacteroides thetaiotaomicron uses the most structurally complex glycan known: the plant pectic polysaccharide rhamnogalacturonan-II, cleaving all but 1 of its 21 distinct glycosidic linkages. The deconstruction of rhamnogalacturonan-II side chains and backbone are coordinated to overcome steric constraints, and the degradation involves previously undiscovered enzyme families and catalytic activities. The degradation system informs revision of the current structural model of rhamnogalacturonan-II and highlights how individual gut bacteria orchestrate manifold enzymes to metabolize the most challenging glycan in the human diet.
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Bacteroides thetaiotaomicron/enzimologia , Bacteroides thetaiotaomicron/metabolismo , Biocatálise , Trato Gastrointestinal/microbiologia , Glicosídeo Hidrolases/metabolismo , Pectinas/química , Pectinas/metabolismo , Bacteroides thetaiotaomicron/crescimento & desenvolvimento , Boratos/química , Boratos/metabolismo , Domínio Catalítico , Microbioma Gastrointestinal , Glicosídeo Hidrolases/química , Glicosídeo Hidrolases/classificação , Humanos , Modelos Moleculares , Especificidade por SubstratoRESUMO
This corrects the article DOI: 10.1038/nature21725.
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BACKGROUND: In fungal plant pathogens, genome rearrangements followed by selection pressure for adaptive traits have facilitated the co-evolutionary arms race between hosts and their pathogens. Pyrenophora tritici-repentis (Ptr) has emerged recently as a foliar pathogen of wheat worldwide and its populations consist of isolates that vary in their ability to produce combinations of different necrotrophic effectors. These effectors play vital roles in disease development. Here, we sequenced the genomes of a global collection (40 isolates) of Ptr to gain insights into its gene content and genome rearrangements. RESULTS: A comparative genome analysis revealed an open pangenome, with an abundance of accessory genes (~ 57%) reflecting Ptr's adaptability. A clear distinction between pathogenic and non-pathogenic genomes was observed in size, gene content, and phylogenetic relatedness. Chromosomal rearrangements and structural organization, specifically around effector coding genes, were detailed using long-read assemblies (PacBio RS II) generated in this work in addition to previously assembled genomes. We also discovered the involvement of large mobile elements associated with Ptr's effectors: ToxA, the gene encoding for the necrosis effector, was found as a single copy within a 143-kb 'Starship' transposon (dubbed 'Horizon') with a clearly defined target site and target site duplications. 'Horizon' was located on different chromosomes in different isolates, indicating mobility, and the previously described ToxhAT transposon (responsible for horizontal transfer of ToxA) was nested within this newly identified Starship. Additionally, ToxB, the gene encoding the chlorosis effector, was clustered as three copies on a 294-kb element, which is likely a different putative 'Starship' (dubbed 'Icarus') in a ToxB-producing isolate. ToxB and its putative transposon were missing from the ToxB non-coding reference isolate, but the homolog toxb and 'Icarus' were both present in a different non-coding isolate. This suggests that ToxB may have been mobile at some point during the evolution of the Ptr genome which is contradictory to the current assumption of ToxB vertical inheritance. Finally, the genome architecture of Ptr was defined as 'one-compartment' based on calculated gene distances and evolutionary rates. CONCLUSIONS: These findings together reflect on the highly plastic nature of the Ptr genome which has likely helped to drive its worldwide adaptation and has illuminated the involvement of giant transposons in facilitating the evolution of virulence in Ptr.
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Ascomicetos , Micotoxinas , Doenças das Plantas/microbiologia , Filogenia , Micotoxinas/genética , Ascomicetos/genéticaRESUMO
This Letter reports the observation of WWW production and a measurement of its cross section using 139 fb^{-1} of proton-proton collision data recorded at a center-of-mass energy of 13 TeV by the ATLAS detector at the Large Hadron Collider. Events with two same-sign leptons (electrons or muons) and at least two jets, as well as events with three charged leptons, are selected. A multivariate technique is then used to discriminate between signal and background events. Events from WWW production are observed with a significance of 8.0 standard deviations, where the expectation is 5.4 standard deviations. The inclusive WWW production cross section is measured to be 820±100 (stat)±80 (syst) fb, approximately 2.6 standard deviations from the predicted cross section of 511±18 fb calculated at next-to-leading-order QCD and leading-order electroweak accuracy.
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Fluorescently labeled, solute-binding proteins that change their fluorescent output in response to ligand binding are frequently used as biosensors for a wide range of applications. We have previously developed a "Computational Identification of Non-disruptive Conjugation sites" (CINC) approach, an in silico pipeline utilizing molecular dynamics simulations for the rapid design and construction of novel protein-fluorophore conjugate-type biosensors. Here, we report an improved in silico scoring algorithm for use in CINC and its use in the construction of an oligogalacturonide-detecting biosensor set. Using both 4,5-unsaturated and saturated oligogalacturonides, we demonstrate that signal transmission from the ligand-binding pocket of the starting protein scaffold to the CINC-selected reporter positions is effective for multiple different ligands. The utility of an oligogalacturonide-detecting biosensor is shown in Carbohydrate Active Enzyme (CAZyme) activity assays, where the biosensor is used to follow product release upon polygalacturonic acid (PGA) depolymerization in real time. The oligogalacturonide-detecting biosensor set represents a novel enabling tool integral to our rapidly expanding platform for biosensor-based carbohydrate detection, and moving forward, the CINC pipeline will continue to enable the rational design of biomolecular tools to detect additional chemically distinct oligosaccharides and other solutes.
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Técnicas Biossensoriais , Corantes Fluorescentes , Ligantes , Oligossacarídeos , ProteínasRESUMO
Gardnerella spp. in the vaginal microbiome are associated with bacterial vaginosis, in which a lactobacillus-dominated community is replaced with mixed bacteria, including Gardnerella species. Co-occurrence of multiple Gardnerella species in the vaginal environment is common, but different species are dominant in different women. Competition for nutrients, including glycogen, could play an important role in determining the microbial community structure. Digestion of glycogen into products that can be taken up and further processed by bacteria requires the combined activities of several enzymes collectively known as amylases, which belong to glycoside hydrolase family 13 (GH13) within the CAZy classification system. GH13 is a large and diverse family of proteins, making prediction of their activities challenging. SACCHARIS annotation of the GH13 family in Gardnerella resulted in identification of protein domains belonging to eight subfamilies. Phylogenetic analysis of predicted amylase sequences from 26 genomes demonstrated that a putative α-glucosidase-encoding sequence, CG400_06090, was conserved in all Gardnerella spp. The predicted α-glucosidase enzyme was expressed, purified, and functionally characterized. The enzyme was active on a variety of maltooligosaccharides with maximum activity at pH 7. Km, kcat, and kcat/Km values for the substrate 4-nitrophenyl α-d-glucopyranoside were 8.3 µM, 0.96 min-1, and 0.11 µM-1 min-1, respectively. Glucose was released from maltose, maltotriose, maltotetraose, and maltopentaose, but no products were detected when the enzyme was incubated with glycogen. Our findings show that Gardnerella spp. produce an α-glucosidase enzyme that may contribute to the multistep process of glycogen metabolism by releasing glucose from maltooligosaccharides. IMPORTANCE Increased abundance of Gardnerella spp. is a diagnostic characteristic of bacterial vaginosis, an imbalance in the human vaginal microbiome associated with troubling symptoms, and negative reproductive health outcomes, including increased transmission of sexually transmitted infections and preterm birth. Competition for nutrients is likely an important factor in causing dramatic shifts in the vaginal microbial community but little is known about the contribution of bacterial enzymes to the metabolism of glycogen, a major carbon source available to vaginal bacteria. The significance of our research is characterizing the activity of an enzyme conserved in Gardnerella species that likely contributes to the ability of these bacteria to utilize glycogen.
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Proteínas de Bactérias/química , Gardnerella/enzimologia , Gardnerella/isolamento & purificação , Microbiota , Vagina/microbiologia , alfa-Glucosidases/química , Sequência de Aminoácidos , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Estabilidade Enzimática , Feminino , Gardnerella/classificação , Gardnerella/genética , Humanos , Concentração de Íons de Hidrogênio , Cinética , Filogenia , Alinhamento de Sequência , Temperatura , alfa-Glucosidases/genética , alfa-Glucosidases/metabolismoRESUMO
The gastrointestinal (GI) tract of humans and animals is lined with mucus that serves as a barrier between the gut microbiota and the epithelial layer of the intestine. As the proteins present in mucus are typically heavily glycosylated, such as the mucins, several enteric commensal and pathogenic bacterial species are well-adapted to this rich carbon source and their genomes are replete with carbohydrate-active enzymes targeted toward dismantling the glycans and proteins present in mucus. One such species is Clostridium perfringens, a Gram-positive opportunistic pathogen indigenous to the gut of humans and animals. The genome of C. perfringens encodes numerous carbohydrate-active enzymes that are predicted or known to target glycosidic linkages within or on the termini of mucus glycans. Through this enzymatic activity, the degradation of the mucosal layer by C. perfringens has been implicated in a number of GI diseases, the most severe of which is necrotic enteritis. In this review, we describe the wide array of extracellular glycoside hydrolases, and their accessory modules, that is possessed by C. perfringens, and examine the unique multimodularity of these proteins in the context of degrading the glycoconjugates in mucus as a potential component of disease.
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Clostridium perfringens , Glicosídeo Hidrolases , Muco , Animais , Glicoconjugados/metabolismo , Glicosídeo Hidrolases/genética , Glicosídeo Hidrolases/metabolismo , Mucinas/metabolismo , Muco/metabolismoRESUMO
The first measurement of longitudinal decorrelations of harmonic flow amplitudes v_{n} for n=2-4 in Xe+Xe collisions at sqrt[s_{NN}]=5.44 TeV is obtained using 3 µb^{-1} of data with the ATLAS detector at the LHC. The decorrelation signal for v_{3} and v_{4} is found to be nearly independent of collision centrality and transverse momentum (p_{T}) requirements on final-state particles, but for v_{2} a strong centrality and p_{T} dependence is seen. When compared with the results from Pb+Pb collisions at sqrt[s_{NN}]=5.02 TeV, the longitudinal decorrelation signal in midcentral Xe+Xe collisions is found to be larger for v_{2}, but smaller for v_{3}. Current hydrodynamic models reproduce the ratios of the v_{n} measured in Xe+Xe collisions to those in Pb+Pb collisions but fail to describe the magnitudes and trends of the ratios of longitudinal flow decorrelations between Xe+Xe and Pb+Pb. The results on the system-size dependence provide new insights and an important lever arm to separate effects of the longitudinal structure of the initial state from other early and late time effects in heavy-ion collisions.
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Several extensions of the Standard Model predict the production of dark matter particles at the LHC. An uncharted signature of dark matter particles produced in association with VV=W^{±}W^{∓} or ZZ pairs from a decay of a dark Higgs boson s is searched for using 139 fb^{-1} of pp collisions recorded by the ATLAS detector at a center-of-mass energy of 13 TeV. The sâV(qq[over ¯])V(qq[over ¯]) decays are reconstructed with a novel technique aimed at resolving the dense topology from boosted VV pairs using jets in the calorimeter and tracking information. Dark Higgs scenarios with m_{s}>160 GeV are excluded.
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A search for charged leptons with large impact parameters using 139 fb^{-1} of sqrt[s]=13 TeV pp collision data from the ATLAS detector at the LHC is presented, addressing a long-standing gap in coverage of possible new physics signatures. Results are consistent with the background prediction. This search provides unique sensitivity to long-lived scalar supersymmetric lepton partners (sleptons). For lifetimes of 0.1 ns, selectron, smuon, and stau masses up to 720, 680, and 340 GeV, respectively, are excluded at 95% confidence level, drastically improving on the previous best limits from LEP.
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A search for lepton-flavor-violating Zâeτ and Zâµτ decays with pp collision data recorded by the ATLAS detector at the LHC is presented. This analysis uses 139 fb^{-1} of Run 2 pp collisions at sqrt[s]=13 TeV and is combined with the results of a similar ATLAS search in the final state in which the τ lepton decays hadronically, using the same data set as well as Run 1 data. The addition of leptonically decaying τ leptons significantly improves the sensitivity reach for Zââτ decays. The Zââτ branching fractions are constrained in this analysis to B(Zâeτ)<7.0×10^{-6} and B(Zâµτ)<7.2×10^{-6} at 95% confidence level. The combination with the previously published analyses sets the strongest constraints to date: B(Zâeτ)<5.0×10^{-6} and B(Zâµτ)<6.5×10^{-6} at 95% confidence level.
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A search for new phenomena is presented in final states with two leptons and one or no b-tagged jets. The event selection requires the two leptons to have opposite charge, the same flavor (electrons or muons), and a large invariant mass. The analysis is based on the full run-2 proton-proton collision dataset recorded at a center-of-mass energy of sqrt[s]=13 TeV by the ATLAS experiment at the LHC, corresponding to an integrated luminosity of 139 fb^{-1}. No significant deviation from the expected background is observed in the data. Inspired by the B-meson decay anomalies, a four-fermion contact interaction between two quarks (b, s) and two leptons (ee or µµ) is used as a benchmark signal model, which is characterized by the energy scale and coupling, Λ and g_{*}, respectively. Contact interactions with Λ/g_{*} lower than 2.0 (2.4) TeV are excluded for electrons (muons) at the 95% confidence level, still far below the value that is favored by the B-meson decay anomalies. Model-independent limits are set as a function of the minimum dilepton invariant mass, which allow the results to be reinterpreted in various signal scenarios.