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| Status |
Public on Jun 10, 2014 |
| Title |
Probing the RyhB-2 regulon in Salmonella Typhimurium |
| Organism |
Salmonella enterica subsp. enterica serovar Typhimurium str. 14028S |
| Experiment type |
Expression profiling by array
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| Summary |
Typically, the expression of sRNAs is activated in response to environmental stimuli in order to regulate gene expression through post-transcriptional mechanisms. In the present work we show that the Salmonella Typhimurium paralog sRNAs RyhB-1 and RyhB-2 are induced in response to the nitrosating agent S-nitrosoglutathione (GSNO). Inactivation of these sRNAs decreased S. Typhimurium resistance to GSNO and increased the levels of nitrosylated proteins. These results prompted us to evaluate a possible role of these sRNAs in nitrosative stress resistance. RNA profiling was used as a screening to identify novel RyhB-1 and RyhB-2 regulated targets, and a subset of genes was filtered based on their potential role in the response to nitrosative stress. To confirm our observation, expression of the candidate targets was analyzed by quantitative RT-PCR in a wild type, single and double mutant strains (ΔryhB1, ΔryhB2 and ΔryhB1 ΔryhB2) treated with GSNO. In response to GSNO RyhB-1 and RyhB-2 negatively regulate the expression of the genes cyoABC (cytochrome o oxidase complex), cydB (cytochrome d oxidase complex), cybC (cytochrome b-562), and positively regulate the nirBCD operon (nitrite reductase system). Together, these results suggest that RyhB-1 and RyhB-2 finely tune the expression of genes coding for cytochrome and the nitrate reductase system, allowing the cell to cope with GSNO-induced stress.
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| Overall design |
Total RNA was harvested from two biological replicates of wt S. Typhimurium overexpressing the sRNA RyhB-2 grown in LB medium, in order to identify RhyB-2 targets that might be implicated in reactive nitrogen stress resistance.
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| Contributor(s) |
Morales EH, Calderon IL, Porwollik S, McClelland M |
| Citation missing |
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| Submission date |
May 29, 2014 |
| Last update date |
Jun 12, 2014 |
| Contact name |
Michael McClelland |
| E-mail(s) |
mcclelland.michael@gmail.com
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| Phone |
858-336-9554
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| Organization name |
University of California, Irvine
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| Department |
Microbiology & Molecular Genetics
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| Street address |
132 Med Surge I
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| City |
Irvine |
| State/province |
CA |
| ZIP/Postal code |
92697 |
| Country |
USA |
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| Platforms (1) |
| GPL14855 |
NIMBLE_MMCCSAL07_14028_380k [MMCCSAL07_100416_SP_CGH] |
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| Samples (2) |
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| Relations |
| BioProject |
PRJNA248921 |
