Signal Pathway in Salt-Activated Expression of theSalmonellaPathogenicity Island 1 Type III Secretion System inSalmonella entericaSerovar Typhimurium

Abstract

Salmonella enterica serovar Typhimurium secretes virulence factors for invasion called Sip proteins or Sips into its hosts through a type III secretion system (T3SS). In the absence of a host, S. enterica induces Sip secretion in response to sucrose or simple salts, such as NaCl. We analyzed induction of host-independent Sip secretion by monitoring protein secretion by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), assembly of needle complexes by electron microscopy, and transcription of virulence regulatory genes by quantitative reverse transcriptase PCR (real-time PCR). SDS-PAGE showed that addition of sucrose or simple salts, such as NaCl, to the growth medium induced Sip secretion without altering flagellar protein secretion, which requires a distinct T3SS. Electron microscopy confirmed that the amount of secreted Sips increased as the number of assembled needle complexes increased. Real-time PCR revealed that added sucrose or NaCl enhanced transcription of hilA, hilC, and hilD, which encode known regulators of Salmonella virulence. However, epistasis analysis implicated HilD and HilA, but not HilC, in the direct pathway from the salt stimulus to the Sip secretion response. Further analyses showed that the BarA/SirA two-component signal transduction pathway, but not the two-component sensor kinase EnvZ, directly activated hilD and hilA transcription and thus Sip secretion in response to either sucrose or NaCl. Finally, real-time PCR showed that salt does not influence transcription of the BarA/SirA-dependent csrB and csrC genes. A model is proposed for the major pathway in which sucrose or salt signals to enhance virulence gene expression.

Flagellar Formation in C-Ring-Defective Mutants by Overproduction of FliI, the ATPase Specific for Flagellar Type III Secretion

“…Osmotically shocked cells were produced from MKM015 (⌬fliN) cells harboring pfliI as described previously (18). In flagellated cells, besides complete flagella, we observed hook-basal body structures sitting on the membranes (Fig.…”

“…Media, culturing, transductional methods, and motility assays were performed as described previously (18).…”

A Genomewide Mutagenesis Screen Identifies Multiple Genes Contributing to Vi Capsular Expression in Salmonella enterica Serovar Typhi

“…The microarray data showed a decrease in viaB locus gene expression of 1 or 2 log, and a reduced but detectable level of Vi polysaccharide expression was detected using an anti-Vi antibody with ELISA. The environmental signals recognized by the barA/sirA system have not been defined fully but are known to include osmolarity (39). BarA and SirA have previously been demonstrated to have a regulatory role in SPI-1 gene expression (39), and we now report another virulence locus, the viaB locus, as part of this regulon (26).…”

Differential expression of Salmonella type III secretion system factors InvJ, PrgJ, SipC, SipD, SopA and SopB in cultures and in mice

“…300 mM NaCl) (Arricau et al, 1998;Huang et al, 2007). However, little is known about the effect of the osmolarity on the protein expression of SPI-1 factors (Mizusaki et al, 2008).…”