On the strong connection between nanoscale adhesion of Yad fimbriae and macroscale attachment of Yad-decorated bacteria to glycosylated, hydrophobic and hydrophilic surfaces.

Yad fimbriae are currently viewed as versatile bacterial adhesins able to significantly mediate host or plant-pathogen recognition and contribute to the persistence of Escherichia coli in both the environment and within hosts. To date, however, the underlying adhesion process of Yad fimbriae on surfaces defined by controlled coating chemistries has not been evaluated on the relevant molecular scale. In this work, the interaction forces operational between Yad fimbriae expressed by genetically modified E. coli and self-assembled monolayers (SAM) differing in terms of charge, hydrophobicity or the nature of decorating sugar units are quantified by Single Molecule Force Spectroscopy (SMFS) on the nanoscale. It is found that the adhesion of Yad fimbriae onto probes functionalized with xylose is as strong as that measured with probes decorated with anti-Yad antibodies (ca. 80 to 300 pN). In contrast, the interactions of Yad with galactose, lactose, mannose, -OH, -NH2, -COOH and -CH3 terminated SAMs are clearly non-specific. Interpretation of SMFS measurements on the basis of worm-like-chain modeling for polypeptide nanomechanics further leads to the estimates of the maximal extension of Yad fimbriae upon stretching, of their persistence length and of their polydispersity. Finally, we show for the first time a strong correlation between the adhesion properties of Yad-decorated bacteria determined from conventional macroscopic counting methods and the molecular adhesion capacity of Yad fimbriae. This demonstration advocates the effort that should be made to understand on the nanoscale level the interactions between fimbriae and their cognate ligands. The results could further help the design of potential anti-adhesive molecules or surfaces to better fight against the virulence of bacterial pathogens.

Drosophila long-term memory formation involves regulation of cathepsin activity.

Whereas short-term memory lasts from minutes to hours, long-term memory (LTM) can last for days or even an entire lifetime. LTM generally forms after spaced repeated training sessions and involves the regulation of gene expression, thereby implicating transcription factors in the initial steps of LTM establishment. However, the direct participation of effector genes in memory formation has been rarely documented, and many of the mechanisms involved in LTM formation remain to be understood. Here we describe a Drosophila melanogaster mutant, crammer (cer), which shows a specific LTM defect. The cer gene encodes an inhibitor of a subfamily of cysteine proteinases, named cathepsins, some of which might be involved in human Alzheimer's disease. The Cer peptide was found in the mushroom bodies (MBs), the Drosophila olfactory memory centre and in glial cells around the MBs. The overexpression of cer in glial cells but not in MB neurons induces a decrease in LTM, suggesting that Cer might have a role in glia and that the concentration of the Cer peptide is critical for LTM. In wild-type flies, cer expression transiently decreases after LTM conditioning, indicating that cysteine proteinases are activated early in LTM formation.

Design and Control of Extrachromosomal Elements in Methylorubrum extorquens AM1.

Genetic tools are a prerequisite to engineer cellular factories for synthetic biology and biotechnology. Methylorubrum extorquens AM1 is an important platform organism of a future C1-bioeconomy. However, its application is currently limited by the availability of genetic tools. Here we systematically tested repABC regions to maintain extrachromosomal DNA in M. extorquens. We used three elements to construct mini-chromosomes that are stably inherited at single copy number and can be shuttled between Escherichia coli and M. extorquens. These mini-chromosomes are compatible among each other and with high-copy number plasmids of M. extorquens. We also developed a set of inducible promoters of wide expression range, reaching levels exceeding those currently available, notably the PmxaF-promoter. In summary, we provide a set of tools to control the dynamic expression and copy number of genetic elements in M. extorquens, which opens new ways to unleash the metabolic and biotechnological potential of this organism for future applications.