A Salmonella typhimurium virulence gene linked to flg.

Isogenic pairs of strains of Salmonella typhimurium which differed only in whether or not they were flagellate were found to be equally virulent in C57BL/6J mice infected orally, intravenously, or intraperitoneally. Therefore, we investigated the genetic basis for our previous observation that in this mouse model, nonflagellate delta flagABCDE25 strains were reduced in virulence compared with isogenic wild-type flagellate strains. The recombinant plasmid pMH6, which contains several flg+ genes and a segment of the S. typhimurium chromosome adjacent to the flg genes, was introduced into a delta flgABCDE25 mutant. This restored virulence in mice challenged intraperitoneally, which suggested that a virulence gene occurs adjacent to the flg genes. When plasmid pMH64, which lacks the chromosomal segment adjacent to the flg genes, was introduced into the same delta flgABCDE25 mutant, virulence was not restored. In contrast, the introduction of pMH71, a plasmid which retains the chromosomal segment adjacent to the flg genes, restored virulence. We concluded that a hitherto unknown virulence gene, which we have named mviS, occurs adjacent to the flg genes and that its absence in delta flgABCDE25 mutants, rather than the nonflagellate phenotype of the delta flgABCDE25 mutants, caused the previously reported attenuation of such mutants.

Salmonella typhimurium virulence in a burned-mouse model.

Various features of salmonellosis were examined in a burned-mouse model. In this model, which uses an outbred mouse strain, a challenge dose of ca. 100 CFU with any of several strains of Salmonella typhimurium caused a fatal infection. A variety of mutated strains attenuated for virulence in Salmonella-susceptible parenterally infected mice were also attenuated in the burned-mouse model. When administered as live vaccines injected intraperitoneally the same attenuated strains provided between slight and complete protection against subsequent lethal challenge subcutaneously at the site of a burn. The correspondence of results obtained in the burned-mouse model with those seen in other mouse models coupled with the unique advantages of the burned-mouse model argue for the usefulness of the model in studies of salmonellosis and in testing of strains constructed for use as live vaccines.

Flagella of Salmonella typhimurium are a virulence factor in infected C57BL/6J mice.

To determine whether flagella, chemotaxis, and motility of Salmonella typhimurium are virulence factors in infected C57BL/6J mice, we constructed isogenic pairs of derivatives of the nonfimbriated virulent strain SL3201. Of each pair, one member contained a mutation in a single gene that is required for expression of normal chemotactically directed motility, whereas the other member contained the wild-type form of the gene. No additional differences between the members of a pair were evident. The phenotypic parameters examined for all derivatives included in vitro growth rate, sensitivity to P22 phage, amino acid auxotrophy, and biotype. For a flagellated and nonflagellated pair, the electron microscopic appearance of each member was examined as well as its lipopolysaccharide and outer membrane profiles by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The virulence of the various derivatives was then assessed in mice challenged orally, intraperitoneally, or intravenously. The results established that flagella, whether functional or nonfunctional as organelles of motility, were S. typhimurium virulence factors and that neither chemotaxis nor motility was required for virulence.

Flagella help Salmonella typhimurium survive within murine macrophages.

In this study, we evaluated how flagella enhance the pathogenicity of Salmonella typhimurium in strain C57BL/6J mice. When mice were infected orally with flagellated or nonflagellated S. typhimurium, equivalent numbers of bacteria colonized the gastrointestinal tracts of the animals, but the number of flagellated organisms increased faster once colonization began in the spleens and livers. To evaluate this differential rate of Salmonella growth, the rate of blood clearance, and the kinetics of net multiplication of salmonellae in splenic tissue after intravenous challenge, the two groups of mice were compared. We found that clearance of bacteria from the blood was the same for flagellated or nonflagellated strains. However, the number of flagellated bacteria in the spleen increased logarithmically until the death of the animals, whereas the number of nonflagellated salmonellae increased only slightly. In contrast, both flagellated and nonflagellated strains grew exponentially in the spleens of mice pretreated with silica, a macrophage toxic agent. In an in vitro macrophage assay, flagellated salmonellae survived longer than nonflagellated organisms. These results indicate that flagella either protect S. typhimurium from the intracellular killing mechanisms of murine macrophages or that flagella enhance the ability of S. typhimurium to multiply within murine macrophages.

Cross-pathway regulation: histidine-mediated control of histidine, tryptophan, and arginine biosynthetic enzymes in Neurospora crassa.

In Neurospora crassa, histidine starvation of histidine mutants resulted in derepression of histidine, tryptophan, and arginine biosynthetic enzymes. The same tripartite derepression occurred in wild-type strain 74A when it was grown in medium supplemented with 3-amino-1,2,4-triazole, an inhibitor of histidine biosynthesis. Histidine-mediated derepression of tryptophan and arginine biosynthetic enzymes was not due to a lowered intracellular concentration of tryptophan or arginine, respectively. A discussion of possible mechanisms and of similar studies in prokaryotic and eukaryotic organisms is presented.

Cross-pathway regulation: tryptophan-mediated control of histidine and arginine biosynthetic enzymes in Neurospora crassa.

In Neurospora crassa, the starvation of tryptophan mutants for tryptophan resulted in the derepression of tryptophan, histidine, and arginine biosynthetic enzymes. This tryptophan-mediated derepression of histidine and arginine biosynthetic enzymes occurred despite the fact that the tryptophan-starved cells had a higher intracellular concentration of histidine and arginine than did nonstarved cells.

Histidine-mediated control of tryptophan biosynthetic enzymes in Neurospora crassa.

The formation of the five tryptophan biosynthetic enzymes of Neurospora crassa was shown to be derepressed in histidine-starved cells. This histidine-mediated derepression was not due to a lowered intracellular concentration of tryptophan in these cells. Furthermore, histidine-mediated derepression of tryptophan enzymes was found to be coordinate and not subject to reversal by tryptophan of either exogenous or biosynthetic origin. The synthesis of tryptophan enzymes also was found to be coordinate in cells which were not histidine-starved. Although histidine is clearly involved in regulating the synthesis of tryptophan enzymes, it did not prevent either tryptophan-mediated derepression of tryptophan enzymes or indole-3-glycerol phosphate-mediated derepression of tryptophan synthetase.