RESUMO
Virtually all bacterial species synthesize high levels of (p)ppGpp (guanosine penta- or tetraphosphate), a pleiotropic regulator of the stringent response and other stresses in bacteria. relA and spoT genes are, respectively, involved in synthesis and synthesis/biodegradation of (p)ppGpp. We aimed in this work to evaluate the impact of static magnetic field (SMF) 200 mT exposure on the expression of relA and spoT genes in Salmonella enterica Hadar. Bacteria were exposed to a SMF during 9 h, and RNA extraction was followed by reverse transcriptase polymerase chain reaction (RT-PCR). The relative quantification of mRNA expression levels using the 16S rRNA reference gene did not change during the SMF exposure. However, results showed a significant increase in gene expression for relA after 3 h of exposure (P < 0.05) and after 6 h for spoT (P < 0.05). The differential gene expression of relA and spoT could be considered as a potential stress response to a SMF exposure in Salmonella related to the production/degradation of (p)ppGpp.
Assuntos
Proteínas de Bactérias , Regulação Bacteriana da Expressão Gênica , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Expressão Gênica , Ligases/genética , Campos Magnéticos , RNA Ribossômico 16S , SalmonellaRESUMO
The present study investigated the adaptation of Salmonella enterica subsp. enterica serovar Hadar to static magnetic field (SMF) exposure (200 mT, 9 h). The proteomic analysis provides an overview of potentially important cytosolic proteins that Salmonella needs to regulate to survive and adapt to magnetic stress. Via 2-dimensional electrophoresis and liquid chromatography tandem mass spectrometry, we compared cytosolic proteomes before and after exposure to magnetic field. A total of 35 proteins displaying more than a 2-fold change were differentially expressed in exposed cells, among which 25 were upregulated and 10 were downregulated. These proteins can be classified mainly into 6 categories: (i) proteins involved in metabolic pathways of carbohydrates, (ii) chaperones and proteins produced in response to oxidative stress, (iii) proteins involved in energy homeostasis, (iv) elongation factors (EF-Tu and EF-Ts), (v) proteins involved in motility, and (vi) proteins involved in molecules transport. Many of the presented observations could be explained, while some represent still-unknown mechanisms. In addition, this study reveals 5 hypothetical proteins. It seems that the stress response to SMF (200 mT) is essentially set up to avoid oxidative damages, with the overexpression of proteins directly involved in oxidative stress response and metabolic switches to counteract oxidative stress. Interestingly, several proteins induced under SMF exposure are found to overlap with those induced by other stresses, such as heat shock and starvation.
Assuntos
Proteínas de Bactérias/metabolismo , Proteoma/metabolismo , Salmonella enterica/metabolismo , Adaptação Fisiológica , Citosol/metabolismo , Metabolismo Energético , Regulação Bacteriana da Expressão Gênica , Campos Magnéticos , Redes e Vias Metabólicas , Chaperonas Moleculares/metabolismo , Fatores de Alongamento de Peptídeos/metabolismo , ProteômicaRESUMO
BACKGROUND: Salmonella enterica serovar Hadar (S. Hadar) is a highly prevalent foodborne pathogen and therefore a major cause of human gastroenteritis worldwide. Outer membrane proteins whose production is often regulated by environmental conditions also play important roles in the adaptability of bacterial pathogens to various environments. RESULTS: The present study investigated the adaptation of S. Hadar under the effect of acute static magnetic field exposure (200 mT, 9 h) and the impact on the outer membrane protein pattern. Via two-dimensional electrophoresis (2-DE) and LC-MS/MS spectrometry, we compared the proteome of enriched-outer membrane fraction before and after exposure to a magnetic field. A total of 11 proteins, displaying more than a two-fold change, were differentially expressed in exposed cells, among which 7 were up-regulated and 4 down-regulated. These proteins were involved in the integrity of cell envelope (TolB, Pal), in the response to oxidative stress (OmpW, dihydrolipoamide dehydrogenase, UspF), in the oxidative stress status (bacterioferritin), in virulence (OmpX, Yfgl) or in motility (FlgE and UspF). Complementary experiments associated the down-regulation of FlgE and UspF with an alteration of swarming, a flagella-driven motility, under SMF. Furthermore, the antibiotic disc diffusion method confirmed a decrease of gentamicin susceptibility in exposed cells. This decrease could be partly associated with the up-regulation of TolC, outer membrane component of an efflux pump. OmpA, a multifunctional protein, was up-regulated. CONCLUSIONS: SMF (200 mT) seems to maintain the cell envelope integrity and to submit the exposed cells to an oxidative stress. Some alterations suggest an increase of the ability of exposed cells to form biofilms.
RESUMO
In the present study, we investigated the effect of exposure to A static magnetic field (SMF) on cell growth, viability, and gene expression of Salmonella enterica subsp. enterica serovar Hadar. Our results indicated that SMF exposure (200 mT, 13 hours) failed to alter cellular growth but induced a decrease of colony-forming units (CFU) between 3 and 6 hours followed by an increase from 6 to 9 hours. The analysis of the differential expression of rpoA, dnaK, katN, and 16S rRNA genes under SMF exposure (200 mT, 10 hours) showed that the expression level of the 16S rRNA mRNA remained stable during the exposure and can thus be used as a reference gene for the analysis on the differential gene expression of Salmonella Hadar. Interestingly, mRNAs of rpoA, katN, and dnaK genes were over-expressed following 10 hours of SMF exposure (200 mT). These data suggest a possible stress response of Salmonella Hadar to static magnetic field.