Structure of the Potential Virulence Factor from Francisella tularensis Shows Unexpected Presence of the SHS2 Motif and Similarity to Other Bacterial Virulence Factors

Pathogenic bacteria attack their host by secreting virulence factors that in various ways interrupt host defenses and damage their cells. Functions of many virulence factors, even from well-studied pathogens, are still unknown. Francisella tularensis is a class A pathogen and a causative agent of tularemia, a disease that is lethal without proper treatment. Here we report the three-dimensional structure and preliminary analysis of the potential virulence factor identified by the transcriptomic analysis of the F. tularensis disease models that is encoded by the FTT_1539 gene. The structure of the FTT_1539 protein contains two sets of three stranded antiparallel beta sheets, with a helix placed between the first and the second beta strand in each sheet. This structural motif, previously seen in virulence factors from other pathogens, was named the SHS2 motif and identified to play a role in protein-protein interactions and small molecule recognition. Sequence and structure analysis identified FTT_1539 as a member of a large family of secreted proteins from a broad range of pathogenic bacteria, such as Helicobacter pylori and Mycobacterium tuberculosis. While the specific function of the proteins from this class is still unknown, their similarity to the H. pylori Tip-α protein that induces TNF-a and other chemokines through NF-kB activation suggests the existence of a common pathogen-host interference mechanism shared by multiple human pathogens.

Production of Poly-γ-Glutamic Acid (γ-PGA) by Clinical Isolates of Staphylococcus Epidermidis

Background and Objective: Poly-γ-glutamic acid (γ-PGA) is a constituent of the Bacillus anthracis capsule and a potential virulence factor of S. epidermidis. In this study, a methodology for the isolation, purification and quantification of γ-PGA in the isolates was adapted. In addition, the fate of the produced γ-PGA and its antiphagocytic activity were investigated. Methods: The capB gene was investigated by the PCR method in 50 isolates of S. epidermidis. A modified methodology was used for the extraction, purification, and quantification of γ-PGA using Cetyltrimethylammonium Bromide (CTAB) solution. The fate of γ-PGA was determined in Tryptic Soy Broth (TSB) medium, as well as the effect of ethanol, NaCl and KCl on the induction of the polymer production. The ability of neutrophils to phagocyte both FITC-labeled latex particles in the presence of free γ-PGA and S. epidermidis with and without anchored γ-PGA was evaluated by cytometry. Results: The production of γ-PGA was detected in 40 isolates; all of them were capB gene carriers. Free γ-PGA was detected and in the strain, the amount of released γ-PGA in the supernatant was 67% greater than the cell anchored γ-PGA. Phagocytosis tests performed with one γ-PGA producer isolate showed a significant reduction in neutrophil internalization. Conclusion: The adapted methodology was able to detect γ-PGA in the isolates studied. In addition to being found attached to the cell wall, it was demonstrated in this study that γ-PGA can also be found in the culture supernatant. Free γ-PGA did not determine a reduction in the internalization of latex by neutrophils, but cells with anchored γ-PGA showed significant protection against phagocytosis.