Propargyl-choline was efficiently incorporated into teichoic acid (TA) polymers on the surface of Streptococcus pneumoniae. The introduction of a fluorophore by click chemistry enabled sufficient labeling of the pneumococcus, as well as its specific detection when mixed with other bacterial species. The labeling is localized at the septal site, suggesting a similar location of the TA and peptidoglycan (PG) synthetic machineries. This method is a unique opportunity to improve our understanding of the spatial location of pneumococcal TA biosynthesis.
Unusual intramolecular cross-links present in adhesins from Gram-positive bacteria have been used to develop a generic process amenable to biotechnology applications. Based on the crystal structure of RrgA, the Streptococcus pneumoniae pilus adhesin, we provide evidence that two engineered protein fragments retain their ability to associate covalently with high specificity, in vivo and in vitro, once isolated from the parent protein. We determined the optimal conditions for the assembly of the complex and we solved its crystal structure at 2 Å. Furthermore, we demonstrate biotechnological applications related to antibody production, nanoassembly and cell-surface labeling based on this process we named Bio Molecular Welding.
Summary
The peptidoglycan is a rigid matrix required to resist turgor pressure and to maintain the cellular shape. It is formed by linear glycan chains composed of N-acetylmuramic acid-(β-1,4)-N-acetylglucosamine (MurNAc-GlcNAc) disaccharides associated through cross-linked peptide stems. The peptidoglycan is continually remodeled by synthetic and hydrolytic enzymes and by chemical modifications, including O-acetylation of MurNAc residues that occurs in most Gram-positive and Gram-negative bacteria. This modification is a powerful strategy developed by pathogens to resist to lysozyme degradation and thus to escape from the host innate immune system but little is known about its physiological function. In this study, we have investigated to what extend peptidoglycan O-acetylation is involved in cell wall biosynthesis and cell division of Streptococcus pneumoniae. We show that O-acetylation driven by Adr protects the peptidoglycan of dividing cells from cleavage by the major autolysin LytA and occurs at the septal site. Our results support a function for Adr in the formation of robust and mature MurNAc O-acetylated peptidoglycan and infer its role in the division of the pneumococcus.
A method for labeling teichoic acids in the human pathogen Streptococcus pneumoniae has been developed using a one-pot two-step metabolic labeling approach. The essential nutriment choline modified with an azido-group was incorporated and exposed at the cell surface more rapidly than it reacted with the strain promoted azide alkyne cycloaddition (SPAAC) partner also present in the medium. Once at the cell surface on teichoic acids, coupling of the azido group could then occur within 5 min by the bio-orthogonal click reaction with a DIBO-linked fluorophore. This fast and easy method allowed pulse-chase experiments and was combined with another fluorescent labeling approach to compare the insertion of teichoic acids with peptidoglycan synthesis with unprecedented temporal resolution. It has revealed that teichoic acid and peptidoglycan processes are largely concomitant, but teichoic acid insertion persists later at the division site.
In Havelange (Belgium), two farms are experiencing an ecological transition. We aimed to evaluate the impact of their agricultural activities on insect pollinator communities. This article depicts the situation at the very early stage of the farm transition. This study supports the fact that the maintenance of farm-level natural habitats provides environmental benefits, such as the conservation of two important pollinator communities: wild bees and hoverflies.
Over two years (2018-2019), by using nets and coloured pan-traps, we collected 6301 bee and hoverfly specimens amongst contrasting habitats within two farmsteads undergoing ecological transition in Havelange (Belgium). We reported 101 bee species and morphospecies from 15 genera within six families and 31 hoverfly species and morphospecies from 18 genera. This list reinforces the national pollinator database by providing new distribution data for extinction-threatened species, such as Andrena schencki Morawitz 1866, Bombus campestris (Panzer 1801), Eucera longicornis (L.) and Halictus maculatus Smith 1848 or for data deficient species, such as A. semilaevis Pérez 1903, A. fulvata (Müller 1766), A. trimmerana (Kirby 1802) and Hylaeus brevicornis Nylander 1852.
Ce travail présente l’inventaire des abeilles sauvages (Hymenoptera Apoidea) observées dans l’ancien quartier Myosotis et le nouveau quartier Engeland de la commune d’Uccle à Bruxelles. Via une campagne d’échantillonnage dans les deux quartiers, 708 spécimens ont été capturés ou observés appartenant à 58 espèces différentes des 6 familles d’abeilles sauvages de Belgique. La faune apiforme capturée représente 14% de la faune belge et 28% de la faune bruxelloise. On y retrouve 10 espèces mono ou oligolectiques et 5 espèces font l’objet d’enjeux de conservation spécifiques selon la liste rouge des abeilles de Belgique. Quelques recommandations de gestion de conservation sont discutées dans ce travail en vue de favoriser au maximum la richesse de ces anthophiles pour la commune d’Uccle.
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