The SdiA-Regulated Gene srgE Encodes a Type III Secreted Effector

Salmonella enterica serovar Typhimurium is a food-borne pathogen that causes severe gastroenteritis. The ability of Salmonella to cause disease depends on two type III secretion systems (T3SSs) encoded in two distinct Salmonella pathogenicity islands, 1 and 2 (SPI1 and SPI2, respectively). S. Typhimurium encodes a solo LuxR homolog, SdiA, which can detect the acyl-homo-serine lactones (AHLs) produced by other bacteria and upregulate the rck operon and the srgE gene. SrgE is predicted to encode a protein of 488 residues with a coiled-coil domain between residues 345 and 382. In silico studies have provided conflicting predictions as to whether SrgE is a T3SS substrate. Therefore, in this work, we tested the hypothesis that SrgE is a T3SS effector by two methods, a -lactamase activity assay and a split green fluorescent protein (GFP) complementation assay. SrgE with -lactamase fused to residue 40, 100, 150, or 300 was indeed expressed and translocated into host cells, but SrgE with -lactamase fused to residue 400 or 488 was not expressed, suggesting interference by the coiled-coil do-main. Similarly, SrgE with GFP S11 fused to residue 300, but not to residue 488, was expressed and translocated into host cells. With both systems, translocation into host cells was dependent upon SPI2. A phylogenetic analysis indicated that srgE is found only within Salmonella enterica subspecies. It is found sporadically within both typhoidal and nontyphoidal sero-vars, although the SrgE protein sequences found within typhoidal serovars tend to cluster separately from those found in non-typhoidal serovars, suggesting functional diversification. The members of the Salmonella genus are rod-shaped, Gram-negative, facultative anaerobes. This genus is divided into tw