σ B-dependent gene induction and expression in Listeria monocytogenes during osmotic and acid stress conditions simulating the intestinal environment

Microbiology ◽  
2004 ◽  
Vol 150 (11) ◽  
pp. 3843-3855 ◽  
Author(s):  
David Sue ◽  
Daniel Fink ◽  
Martin Wiedmann ◽  
Kathryn J. Boor
Keyword(s):  
Listeria Monocytogenes ◽  
Stress Response ◽  
Virulence Genes ◽  
Acid Stress ◽  
Exponential Phase ◽  
Wild Type ◽  
Transcript Accumulation ◽  
Gastrointestinal Infections ◽  
Stress Response Genes ◽  
Foodborne Infections

Listeria monocytogenes must overcome a variety of stress conditions in the host digestive tract to cause foodborne infections. The alternative sigma factor σ B, encoded by sigB, is responsible for regulating transcription of several L. monocytogenes virulence and stress-response genes, including genes that contribute to establishment of gastrointestinal infections. A quantitative RT-PCR assay was used to measure mRNA transcript accumulation for the virulence genes inlA and bsh, the stress-response genes opuCA and lmo0669 (encoding a carnitine transporter and an oxidoreductase, respectively) and the housekeeping gene rpoB. Assays were conducted on mid-exponential phase L. monocytogenes cells exposed to conditions reflecting osmotic (0·3 M NaCl) or acid (pH 4·5) conditions typical for the human intestinal lumen. In exponential-phase cells, as well as under osmotic and acid stress, inlA, opuCA and bsh showed significantly lower absolute expression levels in a L. monocytogenes ΔsigB null mutant compared to wild-type. A statistical model that normalized target gene expression relative to rpoB showed that accumulation of inlA, opuCA and bsh transcripts was significantly increased in the wild-type strain within 5 min of acid and osmotic stress exposure; lmo0669 transcript accumulation increased significantly only after acid exposure. It was concluded that σ B is essential for rapid induction of the tested stress-response and virulence genes under conditions typically encountered during gastrointestinal passage. As inlA, bsh and opuCA are critical for gastrointestinal infections in animal models, the data also suggest that σ B contributes to the ability of L. monocytogenes to cause foodborne infections.

scholarly journals Listeria monocytogenes σB Regulates Stress Response and Virulence Functions

2003 ◽  
Vol 185 (19) ◽  
pp. 5722-5734 ◽  
Author(s):  
Mark J. Kazmierczak ◽  
Sharon C. Mithoe ◽  
Kathryn J. Boor ◽  
Martin Wiedmann
Keyword(s):  
Gene Expression ◽  
Listeria Monocytogenes ◽  
Stress Response ◽  
Consensus Sequence ◽  
Virulence Gene ◽  
Wild Type ◽  
Promoter Regions ◽  
Virulence Gene Expression ◽  
Stress Response Genes ◽  
In The Wild

ABSTRACT While the stress-responsive alternative sigma factor σB has been identified in different species of Bacillus, Listeria, and Staphylococcus, theσ B regulon has been extensively characterized only in B. subtilis. We combined biocomputing and microarray-based strategies to identify σB-dependent genes in the facultative intracellular pathogen Listeria monocytogenes. Hidden Markov model (HMM)-based searches identified 170 candidateσ B-dependent promoter sequences in the strain EGD-e genome sequence. These data were used to develop a specialized, 208-gene microarray, which included 166 genes downstream of HMM-predicted σB-dependent promoters as well as selected virulence and stress response genes. RNA for the microarray experiments was isolated from both wild-type and ΔsigB null mutant L. monocytogenes cells grown to stationary phase or exposed to osmotic stress (0.5 M KCl). Microarray analyses identified a total of 55 genes with statistically significantσ B-dependent expression under the conditions used in these experiments, with at least 1.5-fold-higher expression in the wild type over the sigB mutant under either stress condition (51 genes showed at least 2.0-fold-higher expression in the wild type). Of the 55 genes exhibiting σB-dependent expression, 54 were preceded by a sequence resembling the σB promoter consensus sequence. Rapid amplification of cDNA ends-PCR was used to confirm the σB-dependent nature of a subset of eight selected promoter regions. Notably, theσ B-dependent L. monocytogenes genes identified through this HMM/microarray strategy included both stress response genes (e.g., gadB, ctc, and the glutathione reductase gene lmo1433) and virulence genes (e.g., inlA, inlB, and bsh). Our data demonstrate that, in addition to regulating expression of genes important for survival under environmental stress conditions, σB also contributes to regulation of virulence gene expression in L. monocytogenes. These findings strongly suggest thatσ B contributes to L. monocytogenes gene expression during infection.

Comparison of the Stress Response of Listeria monocytogenes Strains with Sprout Colonization

2008 ◽  
Vol 71 (8) ◽  
pp. 1556-1562 ◽  
Author(s):  
LISA GORSKI ◽  
DENISE FLAHERTY ◽  
JESSICA M. DUHÉ
Keyword(s):  
Oxidative Stress ◽  
Statistical Analysis ◽  
Listeria Monocytogenes ◽  
Stress Response ◽  
Foodborne Pathogen ◽  
Acid Stress ◽  
Broccoli Sprouts ◽  
Alfalfa Sprouts ◽  
Different Strains ◽  
And Oxidative Stress

Twenty-nine strains of the foodborne pathogen Listeria monocytogenes were tested for their ability to colonize alfalfa, radish, and broccoli sprouts and their capacity to withstand acid and oxidative stress, two stresses common to the sprouting environment. Wide variation in the ability of different strains to colonize alfalfa sprouts were confirmed, but the variations among radish and broccoli sprouts were not as large. With a few exceptions, strains that were poor colonizers of alfalfa tended to be among the poorer colonizers of radish and broccoli and vice versa. The strains also were variable in their resistance to both acid and oxidative stress. Statistical analysis revealed no correlation between acid stress and sprout colonization, but there was a positive correlation between resistance to oxidative stress and colonization of all three sprout types. Although the response to oxidative stress is important for L. monocytogenes virulence, it also may be important for life outside of a host.

scholarly journals Phenotypic and Transcriptomic Analyses Demonstrate Interactions between the Transcriptional Regulators CtsR and Sigma B in Listeria monocytogenes

2007 ◽  
Vol 73 (24) ◽  
pp. 7967-7980 ◽  
Author(s):  
Yuewei Hu ◽  
Sarita Raengpradub ◽  
Ute Schwab ◽  
Chris Loss ◽  
Renato H. Orsi ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Stress Response ◽  
Heat Resistance ◽  
Parent Strain ◽  
Inducible Promoter ◽  
Heat Sensitivity ◽  
Intestinal Epithelial ◽  
Class Iii ◽  
Stress Response Genes ◽  
Genes Encoding

ABSTRACT Listeria monocytogenes σB positively regulates the transcription of class II stress response genes; CtsR negatively regulates class III stress response genes. To identify interactions between these two stress response systems, we constructed L. monocytogenes ΔctsR and ΔctsR ΔsigB strains, as well as a ΔctsR strain expressing ctsR in trans under the control of an IPTG (isopropyl-β-d-thiogalactopyranoside)-inducible promoter. These strains, along with a parent and a ΔsigB strain, were assayed for motility, heat resistance, and invasion of human intestinal epithelial cells, as well as by whole-genome transcriptomic and quantitative real-time PCR analyses. Both ΔctsR and ΔctsR ΔsigB strains had significantly higher thermotolerances than the parent strain; however, full heat sensitivity was restored to the ΔctsR strain when ctsR was expressed in trans. Although log-phase ΔctsR was not reduced in its ability to infect human intestinal cells, the ΔctsR ΔsigB strain showed significantly lower invasion efficiency than either the parent strain or the ΔsigB strain, indicating that interactions between CtsR and σB contribute to invasiveness. Statistical analyses also confirmed interactions between the ctsR and the sigB null mutations in both heat resistance and invasion phenotypes. Microarray transcriptomic analyses and promoter searches identified (i) 42 CtsR-repressed genes, (ii) 22 genes with lower transcript levels in the ΔctsR strain, and (iii) at least 40 genes coregulated by both CtsR and σB, including genes encoding proteins with confirmed or plausible roles in virulence and stress response. Our data demonstrate that interactions between CtsR and σB play an important role in L. monocytogenes stress resistance and virulence.

High-Salt Preadaptation of Vibrio parahaemolyticus Enhances Survival in Response to Lethal Environmental Stresses

2014 ◽  
Vol 77 (2) ◽  
pp. 246-253 ◽  
Author(s):  
SAI SIDDARTH KALBURGE ◽  
W. BRIAN WHITAKER ◽  
E. FIDELMA BOYD
Keyword(s):  
Stress Response ◽  
Marine Environment ◽  
Vibrio Parahaemolyticus ◽  
High Salinity ◽  
Acid Stress ◽  
Acid Tolerance ◽  
High Salt ◽  
Wild Type ◽  
Acid Tolerance Response ◽  
Tolerance Response

Adaptation to changing environmental conditions is an important strategy for survival of foodborne bacterial pathogens. Vibrio parahaemolyticus is a gram-negative seafoodborne enteric pathogen found in the marine environment both free living and associated with oysters. This pathogen is a moderate halophile, with optimal growth at 3% NaCl. Among the several stresses imposed upon enteric bacteria, acid stress is perhaps one of the most important. V. parahaemolyticus has a lysine decarboxylase system responsible for decarboxylation of lysine to the basic product cadaverine, an important acid stress response system in bacteria. Preadaptation to mild acid conditions, i.e., the acid tolerance response, enhances survival under lethal acid conditions. Because of the variety of conditions encountered by V. parahaemolyticus in the marine environment and in oyster postharvest facilities, we examined the nature of the V. parahaemolyticus acid tolerance response under high-salinity conditions. Short preadaptation to a 6% salt concentration increased survival of the wild-type strain but not that of a cadA mutant under lethal acid conditions. However, prolonged exposure to high salinity (16 h) increased survival of both the wild-type and the cadA mutant strains. This phenotype was not dependent on the stress response sigma factor RpoS. Although this preadaptation response is much more pronounced in V. parahaemolyticus, this characteristic is not limited to this species. Both Vibrio cholerae and Vibrio vulnificus also survive better under lethal acid stress conditions when preadapted to high-salinity conditions. High salt both protected the organism against acid stress and increased survival under −20°C cold stress conditions. High-salt adaptation of V. parahaemolyticus strains significantly increases survival under environmental stresses that would otherwise be lethal to these bacteria.

scholarly journals Strain dependent expression of stress response and virulence genes of Listeria monocytogenes in meat juices as determined by microarray

2012 ◽  
Vol 152 (3) ◽  
pp. 116-122 ◽  
Author(s):  
Kalliopi Rantsiou ◽  
Anna Greppi ◽  
Matteo Garosi ◽  
Alberto Acquadro ◽  
Marios Mataragas ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Stress Response ◽  
Virulence Genes ◽  
Strain Dependent

scholarly journals The bvr Locus of Listeria monocytogenes Mediates Virulence Gene Repression by β-Glucosides

1999 ◽  
Vol 181 (16) ◽  
pp. 5024-5032 ◽  
Author(s):  
Klaus Brehm ◽  
María-Teresa Ripio ◽  
Jürgen Kreft ◽  
José-Antonio Vázquez-Boland
Keyword(s):  
Listeria Monocytogenes ◽  
Virulence Genes ◽  
Natural Habitat ◽  
Sensory System ◽  
Virulence Gene ◽  
Gene Repression ◽  
Signal Molecules ◽  
Wild Type ◽  
Phosphotransferase System ◽  
Virulence Gene Expression

ABSTRACT The β-glucoside cellobiose has been reported to specifically repress the PrfA-dependent virulence genes hly andplcA in Listeria monocytogenes NCTC 7973. This led to the hypothesis that β-glucosides, sugars of plant origin, may act as signal molecules, preventing the expression of virulence genes if L. monocytogenes is living in its natural habitat (soil). In three other laboratory strains (EGD, L028, and 10403S), however, the effect of cellobiose was not unique, and all fermentable carbohydrates repressed hly. This suggested that the downregulation of virulence genes by β-glucosides is not a specific phenomenon but, rather, an aspect of a global regulatory mechanism of catabolite repression (CR). We assessed the effect of carbohydrates on virulence gene expression in a panel of wild-type isolates of L. monocytogenes by using the PrfA-dependent phospholipase C geneplcB as a reporter. Utilization of any fermentable sugar caused plcB repression in wild-type L. monocytogenes. However, an EGD variant was identified in which, as in NCTC 7973, plcB was only repressed by β-glucosides. Thus, the regulation of L. monocytogenes virulence genes by sugars appears to be mediated by two separate mechanisms, one presumably involving a CR pathway and another specifically responding to β-glucosides. We have identified in L. monocytogenes a 4-kb operon, bvrABC, encoding an antiterminator of the BglG family (bvrA), a β-glucoside-specific enzyme II permease component of the phosphoenolpyruvate-sugar phosphotransferase system (bvrB), and a putative ADP-ribosylglycohydrolase (bvrC). Low-stringency Southern blots showed that this locus is absent from other Listeria spp. Transcription ofbvrB was induced by cellobiose and salicin but not by arbutin. Disruption of the bvr operon by replacing part ofbvrAB with an interposon abolished the repression by cellobiose and salicin but not that by arbutin. Our data indicate that the bvr locus encodes a β-glucoside-specific sensor that mediates virulence gene repression upon detection of cellobiose and salicin. Bvr is the first sensory system found in L. monocytogenes that is involved in environmental regulation of virulence genes.

scholarly journals THE ROLE OF TRANSCRIPTION FACTORS DFOXO, DSIR2 AND HSP70 IN LIFESPAN ALTERATION OF DROSOPHILA MELANOGASTER IN DIFFERENT LIGHT CONDITIONS

2010 ◽  
Vol 8 (3) ◽  
pp. 67-80 ◽  
Author(s):  
Aleksey A Moskalev ◽  
Olga A Malysheva
Keyword(s):  
Drosophila Melanogaster ◽  
Transcription Factors ◽  
Stress Response ◽  
Life Span ◽  
Light Regime ◽  
Light Conditions ◽  
Wild Type ◽  
Stress Response Genes ◽  
Significant Difference

It was investigated the role of stress-response genes (dFOXO, dSir2, Hsp70) in regulation of life span of Drosophila in response to light regime alteration. It was revealed the FOXO-dependant mechanism of lifespan increasing at darkness conditions. The distance of lifespan of FOXO homozygous mutants at different light conditions were absent 3 times from 4 times. It was shown, that homozygotes with deletion of dSir2 have more significant difference between lifespan at standard light and darkness conditions with comparing to wild type and heterozygous strain. The same tendency was also detected the in the strains with Hsp70 deletions. It was produced the evidences of two mechanisms of light regime influence on lifespan: metabolism intensification at light conditions and neuroendocrine-determinated lifespan increasing at darkness conditions.

scholarly journals Comparative Analysis of the σB-Dependent Stress Responses in Listeria monocytogenes and Listeria innocua Strains Exposed to Selected Stress Conditions

2007 ◽  
Vol 74 (1) ◽  
pp. 158-171 ◽  
Author(s):  
Sarita Raengpradub ◽  
Martin Wiedmann ◽  
Kathryn J. Boor
Keyword(s):  
Listeria Monocytogenes ◽  
Stress Response ◽  
Stationary Phase ◽  
Stress Responses ◽  
Bacterial Species ◽  
Acid Stress ◽  
Listeria Innocua ◽  
Transcript Levels ◽  
Genes Encoding ◽  
Flagellar Proteins

ABSTRACT The alternative sigma factor σB contributes to transcription of stress response and virulence genes in diverse gram-positive bacterial species. The composition and functions of the Listeria monocytogenes and Listeria innocua σB regulons were hypothesized to differ due to virulence differences between these closely related species. Transcript levels in stationary-phase cells and in cells exposed to salt stress were characterized by microarray analyses for both species. In L. monocytogenes, 168 genes were positively regulated by σB; 145 of these genes were preceded by a putative σB consensus promoter. In L. innocua, 64 genes were positively regulated by σB. σB contributed to acid stress survival in log-phase cells for both species but to survival in stationary-phase cells only for L. monocytogenes. In summary, (i) the L. monocytogenes σB regulon includes >140 genes that are both directly and positively regulated by σB, including genes encoding proteins with importance in stress response, virulence, transcriptional regulation, carbohydrate metabolism, and transport; (ii) a number of L. monocytogenes genes encoding flagellar proteins show higher transcript levels in the ΔsigB mutant, and both L. monocytogenes and L. innocua ΔsigB null mutants have increased motility compared to the respective isogenic parent strains, suggesting that σB affects motility and chemotaxis; and (iii) although L. monocytogenes and L. innocua differ in σB-dependent acid stress resistance and have species-specific σB-dependent genes, the L. monocytogenes and L. innocua σB regulons show considerable conservation, with a common set of at least 49 genes that are σB dependent in both species.

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