Effect of E-beam treatment on expression of virulence and stress-response genes of Listeria monocytogenes in dry-cured ham

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.

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Listeria monocytogenes σB Regulates Stress Response and Virulence Functions

Journal of Bacteriology ◽

10.1128/jb.185.19.5722-5734.2003 ◽

2003 ◽

Vol 185 (19) ◽

pp. 5722-5734 ◽

Cited By ~ 246

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.

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

Microbiology ◽

10.1099/mic.0.27257-0 ◽

2004 ◽

Vol 150 (11) ◽

pp. 3843-3855 ◽

Cited By ~ 123

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.

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Characterization of new stress response genes in Listeria monocytogenes

Free Radical Biology and Medicine ◽

10.1016/j.freeradbiomed.2018.04.498 ◽

2018 ◽

Vol 120 ◽

pp. S151

Author(s):

Sofia Boavida ◽

Ricardo dos Santos ◽

Ana Quendera ◽

Cecilia Arraiano ◽

José Andrade

Keyword(s):

Listeria Monocytogenes ◽

Stress Response ◽

Stress Response Genes

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Transcriptomic and Phenotypic Analyses Suggest a Network between the Transcriptional Regulators HrcA and σB in Listeria monocytogenes

Applied and Environmental Microbiology ◽

10.1128/aem.01281-07 ◽

2007 ◽

Vol 73 (24) ◽

pp. 7981-7991 ◽

Cited By ~ 40

Author(s):

Yuewei Hu ◽

Haley F. Oliver ◽

Sarita Raengpradub ◽

M. Elizabeth Palmer ◽

Renato H. Orsi ◽

...

Keyword(s):

Listeria Monocytogenes ◽

Stress Response ◽

Heat Resistance ◽

Acid Resistance ◽

Transcriptional Regulators ◽

Intestinal Epithelial ◽

Quantitative Real Time Pcr ◽

Stress Response Genes ◽

Human Intestinal Epithelial Cells ◽

Number Of Genes

ABSTRACT Listeria monocytogenes HrcA and CtsR negatively regulate class I and III stress response genes, respectively, while σB positively regulates the transcription of class II stress response genes. To define the HrcA regulon and identify interactions between HrcA, CtsR, and σB, we characterized newly generated L. monocytogenes ΔhrcA, ΔctsR ΔhrcA, and ΔhrcA ΔsigB strains, along with previously described ΔsigB, ΔctsR, and ΔctsR ΔsigB strains, using phenotypic assays (i.e., heat resistance, acid resistance, and invasion of human intestinal epithelial cells) and performed whole-genome transcriptome analysis of the ΔhrcA strain. The hrcA and sigB deletions had significant effects on heat resistance. While the hrcA deletion had no significant effect on acid resistance or invasion efficiency in Caco-2 cells, a linear regression model revealed a significant (P = 0.0493) effect of interactions between the hrcA deletion and the ctsR deletion on invasiveness. Microarray-based transcriptome analyses and promoter searches identified (i) 25 HrcA-repressed genes, including two operons (the groESL and dnaK operons, both confirmed as HrcA regulated by quantitative real-time PCR) and one gene directly repressed by HrcA, and (ii) 36 genes that showed lower transcript levels in the ΔhrcA strain and thus appear to be indirectly upregulated by HrcA. A number of genes were found to be coregulated by either HrcA and CtsR (2 genes), HrcA and σB (31 genes), or all three regulators (5 genes, e.g., gadCB). Combined with previous evidence that σB appears to directly regulate hrcA transcription, our data suggest that HrcA and σB, as well as CtsR, form a regulatory network that contributes to the transcription of a number of L. monocytogenes genes.

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AITRL, an evolutionarily conserved plant specific transcription repressor regulates ABA response in Arabidopsis

Scientific Reports ◽

10.1038/s41598-020-80695-2 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Yanxing Ma ◽

Hainan Tian ◽

Rao Lin ◽

Wei Wang ◽

Na Zhang ◽

...

Keyword(s):

Stress Response ◽

Protein Phosphatase ◽

Aba Signaling ◽

Response Gene ◽

Stress Response Genes ◽

Arabidopsis Protein ◽

Evolutionarily Conserved ◽

Increased Sensitivity ◽

Aba Response ◽

Transcription Repressors

AbstractExpression of stress response genes can be regulated by abscisic acid (ABA) dependent and ABA independent pathways. Osmotic stresses promote ABA accumulation, therefore inducing the expression of stress response genes via ABA signaling. Whereas cold and heat stresses induce the expression of stress response genes via ABA independent pathway. ABA induced transcription repressors (AITRs) are a family of novel transcription factors that play a role in ABA signaling, and Drought response gene (DRG) has previously been shown to play a role in regulating plant response to drought and freezing stresses. We report here the identification of DRG as a novel transcription factor and a regulator of ABA response in Arabidopsis. We found that the expression of DRG was induced by ABA treatment. Homologs searching identified AITR5 as the most closely related Arabidopsis protein to DRG, and homologs of DRG, including the AITR-like (AITRL) proteins in bryophytes and gymnosperms, are specifically presented in embryophytes. Therefore we renamed DRG as AITRL. Protoplast transfection assays show that AITRL functioned as a transcription repressor. In seed germination and seedling greening assays, the aitrl mutants showed an increased sensitivity to ABA. By using qRT-PCR, we show that ABA responses of some ABA signaling component genes including some PYR1-likes (PYLs), PROTEIN PHOSPHATASE 2Cs (PP2Cs) and SUCROSE NONFERMENTING 1 (SNF1)-RELATED PROTEIN KINASES 2s (SnRK2s) were reduced in the aitrl mutants. Taken together, our results suggest that AITRLs are a family of novel transcription repressors evolutionally conserved in embryophytes, and AITRL regulates ABA response in Arabidopsis by affecting ABA response of some ABA signaling component genes.

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Transcriptional response of key metabolic and stress response genes of a nuculanid bivalve, Lembulus bicuspidatus from an oxygen minimum zone exposed to hypoxia-reoxygenation

Comparative Biochemistry and Physiology Part B Biochemistry and Molecular Biology ◽

10.1016/j.cbpb.2021.110617 ◽

2021 ◽

pp. 110617

Author(s):

Katherine Amorim ◽

Helen Piontkivska ◽

Michael L. Zettler ◽

Eugene Sokolov ◽

Tjorven Hinzke ◽

...

Keyword(s):

Stress Response ◽

Transcriptional Response ◽

Oxygen Minimum Zone ◽

Stress Response Genes ◽

Minimum Zone ◽

Oxygen Minimum ◽

Hypoxia Reoxygenation

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Polymorphisms in stress response genes in Lactobacillus plantarum: implications for classification and heat stress response

Annals of Microbiology ◽

10.1007/s13213-014-0862-7 ◽

2014 ◽

Vol 65 (1) ◽

pp. 297-305

Author(s):

Angela Guidone ◽

Eugenio Parente ◽

Teresa Zotta ◽

Caitriona M. Guinane ◽

Mary C. Rea ◽

...

Keyword(s):

Heat Stress ◽

Stress Response ◽

Lactobacillus Plantarum ◽

Heat Stress Response ◽

Stress Response Genes

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Vitamin D regulates the expression of immune and stress response genes in dengue virus-infected macrophages by inducing specific microRNAs

MicroRNA ◽

10.2174/2211536610666211221151949 ◽

2021 ◽

Vol 11 ◽

Author(s):

Geysson Javier Fernandez ◽

Jorge Andrés Castillo ◽

Diana Marcela Giraldo ◽

Silvio Urcuqui-Inchima

Keyword(s):

Vitamin D ◽

Immune Response ◽

Stress Response ◽

Dengue Virus ◽

Mirna Expression ◽

Expression Profiles ◽

Denv Infection ◽

High Dose ◽

Regulatory Pathways ◽

Stress Response Genes

Background: The pathogenesis associated with Dengue virus (DENV) infection is marked by the impairment of host immune response. Consequently, the modulation of immune response has emerged as an important therapeutic target for the control of DENV infection. Vitamin D has been shown to regulate the immune response in DENV infection, although the molecular mechanism remains poorly understood. Post-transcriptional regulation of mRNA by miRNAs offers an opportunity to gain insight into the immunomodulation mediated by vitamin D Objective: Previously, it has been observed that a high dose of vitamin D (4000 IU) decreased DENV-2 infection and inflammatory response in monocyte-derived macrophages (MDMs). Here, we examine whether high or low doses of vitamin D supplements exert differential effect on miRNA expression in DENV-infected macrophages Methods: We analyzed miRNA expression profiles in MDMs isolated from healthy individuals who were given either 1000 or 4000 IU/day of vitamin D for 10 days. MDMs before or after vitamin D supplementation were challenged with DENV-2, and miRNAs profiles were analyzed by qPCR arrays. Results: DENV-2 infected MDMs supplemented with 4000 IU, showed up-regulation of miR-374a-5p, miR-363-3p, miR-101-3p, miR-9-5p, miR-34a-5p, miR-200a-3p, and the family of miRNAs miR-21-5p, and miR-590-p. The miRNA profile and predicted target mRNAs suggested regulatory pathways in MDMs obtained from healthy donors who received higher doses of vitamin D. These DENV-2 infected MDMs expressed a unique set of miRNAs that target immune and cellular stress response genes. Conclusion: The results suggest vitamin D dose-dependent differential expression of miRNAs target key signaling pathways of the pathogenesis of dengue disease.

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