Contribution of SPI-4 genes to the virulence of Salmonella enterica.

Salmonella pathogenicity island-4 (SPI-4) is a 27-kb region that carries six genes designated siiABCDEF. SiiC, SiiD, and SiiF form a type I secretion apparatus for the secretion of SiiE, a huge (approximately 600 kDa) protein contributing to the colonization of the bovine intestines. Here it is shown that loss of SPI-4 attenuates the oral virulence of Salmonella enterica serovars Typhimurium and Enteritidis in mice. Fifty percent lethal doses were elevated in both serovars upon the loss of SPI-4. Moreover, delta SPI-4 mutants were outcompeted in systemic organs by their wild-type strains in a cochallenge model. Contribution of SPI-4 to virulence appeared less pronounced in the S. Enteritidis strain, which was justified by lower levels of the secreted protein SiiE in this strain in comparison with S. Typhimurium. Competition assays with isogenic mutants lacking individual genes of the island showed that all six genes were required for full virulence of S. Typhimurium. Delta siiA and delta siiB mutants were, nevertheless, able to secrete SiiE to culture supernatants. The amount of secreted SiiE was, however, reduced in these two mutants compared with the wild-type strain. Furthermore, a down-regulation of SiiE levels is shown in structural and regulatory lipopolysaccharide mutants exhibiting the deep-rough phenotype.