Initial Characterization of the FlgE Hook High Molecular Weight Complex of

The spirochete periplasmic flagellum has many unique attributes. One unusual characteristic is the flagellar hook. This structure serves as a universal joint coupling rotation of the membrane-bound motor to the flagellar filament. The hook is comprised of about 120 FlgE monomers, and in most bacteria these structures readily dissociate to monomers (∼ 50 kDa) when treated with heat and detergent. However, in spirochetes the FlgE monomers form a large mass of over 250 kDa [referred to as a high molecular weight complex (HMWC)] that is stable to these and other denaturing conditions. In this communication, we examined specific aspects with respect to the formation and structure of this complex. We found that the Lyme disease spirochete Borrelia burgdorferi synthesized the HMWC throughout the in vitro growth cycle, and also in vivo when implanted in dialysis membrane chambers in rats. The HMWC was stable to formic acid, which supports the concept that the stability of the HMWC is dependent on covalent cross-linking of individual FlgE subunits. Mass spectrometry analysis of the HMWC from both wild type periplasmic flagella and polyhooks from a newly constructed ΔfliK mutant indicated that other proteins besides FlgE were not covalently joined to the complex, and that FlgE was the sole component of the complex. In addition, mass spectrometry analysis also indicated that the HMWC was composed of a polymer of the FlgE protein with both the N- and C-terminal regions remaining intact. These initial studies set the stage for a detailed characterization of the HMWC. Covalent cross-linking of FlgE with the accompanying formation of the HMWC we propose strengthens the hook structure for optimal spirochete motility

Landscape level processes driving carabid crop assemblage in dynamic farmlands

International audienceLandscape heterogeneity has been shown to be a major factor in the maintenance of biodiversity and associated services in agricultural landscapes. Farmlands are mosaics of fields with various crop types and farming practices. Crop phenology creates asynchrony between fields sown and harvested in different periods (winter vs. spring crops). The present study was conducted to examine the influence of such spatio-temporal heterogeneity on biodiversity, with the hypothesis that it would lead to spatio-temporal redistribution (shifting) of species. Species richness and activity-density of carabid beetles in winter cereal (winter) and maize (spring) crops were compared across 20 landscapes distributed along a double gradient of relative area and spatial configuration of winter and spring crops. Maize fields were sampled in spring and late summer for comparison over time. The response of carabid species richness to landscape heterogeneity was weak in spring, but maize field richness benefited from adjacencies with woody habitat, in late summer. In spring, increased length of interfaces between winter and spring crops lowered carabid activity-density in winter cereal fields, suggesting that maize fields acted as sinks. Interfaces between woody habitats and crops increased activity-density in both crop types. We found no evidence of spatio-temporal complementation, but different species benefited from winter cereals and maize in spring and late summer, increasing overall diversity. These findings confirm the role of adjacencies between woody and cultivated habitats in the conservation of abundant carabid assemblage in winter cereals and maize. We conclude that between-field population movement occurs, and advocate for better consideration of farmland heterogeneity in future researc

High-avidity IgA protects the intestine by enchaining growing bacteria

Vaccine-induced high-avidity IgA can protect against bacterial enteropathogens by directly neutralizing virulence factors or by poorly defined mechanisms that physically impede bacterial interactions with the gut tissues ('immune exclusion'). IgA-mediated cross-linking clumps bacteria in the gut lumen and is critical for protection against infection by non-typhoidal Salmonella enterica subspecies enterica serovar Typhimurium (S. Typhimurium). However, classical agglutination, which was thought to drive this process, is efficient only at high pathogen densities (≥10; 8; non-motile bacteria per gram). In typical infections, much lower densities (10; 0; -10; 7; colony-forming units per gram) of rapidly dividing bacteria are present in the gut lumen. Here we show that a different physical process drives formation of clumps in vivo: IgA-mediated cross-linking enchains daughter cells, preventing their separation after division, and clumping is therefore dependent on growth. Enchained growth is effective at all realistic pathogen densities, and accelerates pathogen clearance from the gut lumen. Furthermore, IgA enchains plasmid-donor and -recipient clones into separate clumps, impeding conjugative plasmid transfer in vivo. Enchained growth is therefore a mechanism by which IgA can disarm and clear potentially invasive species from the intestinal lumen without requiring high pathogen densities, inflammation or bacterial killing. Furthermore, our results reveal an untapped potential for oral vaccines in combating the spread of antimicrobial resistance