Modified intracellular-associated phenotypes in a recombinant Salmonella Typhi expressing S. Typhimurium SPI-3 sequences.
A bioinformatics comparison of Salmonella Pathogenicity Island 3 sequences from S. Typhi and S. Typhimurium serovars showed that ten genes are highly conserved. However three of them are pseudogenes in S. Typhi. Our aim was to understand what functions are lost in S. Typhi due to pseudogenes by constructing a S. Typhi genetic hybrid carrying the SPI-3 region of S. Typhimurium instead of its own SPI-3. We observed that under stressful conditions the hybrid strain showed a clear impairment in resistance to hydrogen peroxide and decreased survival within U937 culture monocytes. We hypothesized that the marT-fidL operon, encoded in SPI-3, was responsible for the new phenotypes because marT is a pseudogen in S. Typhi and has a demonstrated role as a transcriptional regulator in S. Typhimurium. Therefore we cloned and transferred the S. Typhimurium marT-fidL operon into S. Typhi and confirmed that invasion of monocytes was dramatically decreased. Finally, our findings suggest that the genomic and functional differences between SPI-3 sequences have implications in the host specificity of Typhi and Typhimurium serovars.
Retamal
Patricio
P
Departamento de Medicina Preventiva Animal, Universidad de Chile, Santiago, Chile.
Castillo-Ruiz
Mario
M
Villagra
Nicolás A
NA
Morgado
Juan
J
Mora
Guido C
GC
D000693:Anaerobiosis; D015964:Gene Expression Regulation, Bacterial; D023061:Gene Order; D005798:Genes, Bacterial / Q000235:genetics; D044404:Genomic Islands / Q000235:genetics*; D005838:Genotype; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D050296:Microbial Viability / Q000187:drug effects / Q000235:genetics*; D009154:Mutation; D009876:Operon / Q000235:genetics; D010641:Phenotype; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012485:Salmonella typhi / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity; D012486:Salmonella typhimurium / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity; D013696:Temperature; D014170:Transformation, Genetic; D020298:U937 Cells
D006861:Hydrogen Peroxide