EF-P dependent genes in Salmonella Typhimurium
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ABSTRACT:
PROVIDER: PRJNA1050244 | ENA |
REPOSITORIES: ENA
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Project description:Elongation factor P (EF-P) is required to enhance peptidyl transferase on pausing ribosomes, which can be induced by certain codons on mRNA. It regulates expression of genes in specific conditions. Therefore, to search EF-P mediated ribosome stalling motif, ribosome profiling has been used on organisms such as yeast and E. coli. However, it has not been addressed on the intracellular pathogen Salmonella Typhimurium. Through the ribosome profiling, 135 sites in 128 genes were identified as EF-P dependent sequences and many of these sites had consecutive proline codons specifically at P site of the ribosome. As a motif level, PPG was the most abundant amino acid sequence at EPA site of the ribosome and followed by APP and DPP sequence. Also, our findings revealed that salmonella-specific gene, ugtL, is regulated by frameshifting caused during translation as ribosome stalled in the DPP motif. In addition, we showed that both proline motif and EF-P are engaged in this regulation, and it affects the resistance to antimicrobial peptides. In conclusion, this study emphasized the importance of poly proline motif on EF-P and suggested a possible mechanism how EF-P affects the physiology of Salmonella.
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Project description:Salmonella enterica serotype Typhimurium produces a variety of fimbrial appendages, among which the type 1 fimbriae is the most common type. In vitro static broth culture favors S. Typhimurium to produce type 1 fimbriae, while solid agar inhibits its expression. A transposon inserted in the stbC gene, which would encode an usher protein for Stb fimbriae of a non-flagellar S. Typhimurium LB5010 strain, conferred it to agglutinate yeast cells on both cultures, and was mannose-sensitive. Reverse transcription polymerase chain reaction (RT-PCR) revealed that the expression of the fimbrial major subunit gene fimA, and fimZ, a positive regulator gene of fimA, were both increased in the stbC mutant strain when grown on LB agar; fimW, a repressor gene of fimA, exhibited lower expression. Flagella were observed in the stbC mutant and this phenotype was correlated with the motile phenotype detected by MSRV agar medium and reaction with flagella antiserum. Microarray data and RT-PCR also indicated that the expression of three genes, motA, motB, and cheM, was enhanced in the stbC mutant. The S. Typhimurium stbC mutant was resistant to a variety of antibiotics, consistent with the finding that expression of yhcQ and ramA, two genes involved in multidrug resistance, was enhanced. A complementation test revealed that transforming a recombinant plasmid possessing the coding sequence of the stbC gene restored the mannose-sensitive agglutination phenotype to the stbC mutant much as that in the parental S. Typhimurium LB5010 strain, indicating the possibility of an interplay of different fimbrial systems in coordinating their expression. Key Words: Salmonella enterica serotype Typhimurium, fimbriae, type 1 fimbriae, stbC, transposon, multidrug resistant, flagella
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