title: Spa15 of Shigella flexneri is secreted through the type III secretion system and prevents staurosporine-induced apoptosis.
abstract: Shigella flexneri is a gram-negative, facultative intracellular pathogen that invades the colonic epithelium and causes bacillary dysentery. We previously demonstrated that S. flexneri inhibits staurosporine-induced apoptosis in infected epithelial cells and that a DeltamxiE mutant is unable to inhibit apoptosis. Therefore, we hypothesized that an MxiE-regulated gene was responsible for protection of epithelial cells from apoptosis. Analysis of all MxiE-regulated genes yielded no mutants that lacked the ability to prevent apoptosis. Spa15, which is defined as a type III secretion system chaperone, was analyzed since it associates with MxiE. A Deltaspa15 mutant was unable to prevent staurosporine-induced apoptosis. C-terminal hemagglutinin-tagged spa15 was secreted by S. flexneri within 2 h in the Congo red secretion assay, and secretion was dependent on the type III secretion system. Spa15 was also secreted by Shigella in infected epithelial cells, as verified by immunofluorescence analysis. Spa15 secretion was decreased in the DeltamxiE mutant, which demonstrates why this mutant is unable to prevent staurosporine-induced apoptosis. Our data are the first to show that Spa15 is secreted in a type III secretion system-dependent fashion, and the absence of Spa15 in the Deltaspa15 mutant results in the loss of protection from staurosporine-induced apoptosis in epithelial cells. Thus, Spa15 contributes to the intracellular survival of Shigella by blocking apoptosis in the infected host cell.
authors.affiliation: Department of Microbiology and Immunology, F Edward Hébert School of Medicine, Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814-4799, USA.
authors.forename: Christina S
authors.initials: CS
authors.lastname: Faherty
authors.forename: Anthony T
authors.initials: AT
authors.lastname: Maurelli
mesh_terms: D017209:Apoptosis; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004847:Epithelial Cells / Q000382:microbiology; D017353:Gene Deletion; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism*; D012763:Shigella flexneri / Q000235:genetics / Q000472:pathogenicity*; D019311:Staurosporine / Q000037:antagonists & inhibitors* / Q000633:toxicity*
chemical_list: D001426:Bacterial Proteins; D018832:Molecular Chaperones; C461380:Spa15 protein, Shigella flexneri; D019311:Staurosporine