scholarly journals Role of Cold Shock Proteins in Growth of Listeria monocytogenes under Cold and Osmotic Stress Conditions

2009 ◽  
Vol 75 (6) ◽  
pp. 1621-1627 ◽  
Author(s):  
Barbara Schmid ◽  
Jochen Klumpp ◽  
Eveline Raimann ◽  
Martin J. Loessner ◽  
Roger Stephan ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Osmotic Stress ◽  
Cold Shock ◽  
Microbial Control ◽  
Nacl Stress ◽  
Stress Conditions ◽  
Control Measures ◽  
Cold Shock Protein ◽  
Osmotic Stress Tolerance ◽  
Shock Proteins

ABSTRACT The gram-positive bacterium Listeria monocytogenes is a food-borne pathogen of both public health and food safety significance. It possesses three small, highly homologous protein members of the cold shock protein (Csp) family. We used gene expression analysis and a set of mutants with single, double, and triple deletions of the csp genes to evaluate the roles of CspA, CspB, and CspD in the cold and osmotic (NaCl) stress adaptation responses of L. monocytogenes. All three Csps are dispensable for growth at optimal temperature (37°C). These proteins are, however, required for efficient cold and osmotic stress tolerance of this bacterium. The hierarchies of their functional importance differ, depending on the environmental stress conditions: CspA>CspD>CspB in response to cold stress versus CspD>CspA/CspB in response to NaCl salt osmotic stress. The fact that Csps are promoting L. monocytogenes adaptation against both cold and NaCl stress has significant implications in view of practical food microbial control measures. The combined or sequential exposure of L. monocytogenes cells to these two stresses in food environments might inadvertently induce cross-protection responses.

scholarly journals Listeria monocytogenes Cold Shock Proteins: Small Proteins with A Huge Impact

2021 ◽  
Vol 9 (5) ◽  
pp. 1061
Author(s):  
Francis Muchaamba ◽  
Roger Stephan ◽  
Taurai Tasara
Keyword(s):  
Listeria Monocytogenes ◽  
Stress Tolerance ◽  
Cold Shock ◽  
Current Data ◽  
Health Concern ◽  
Stress Exposure ◽  
Cold Shock Proteins ◽  
Cell Functions ◽  
Small Proteins ◽  
Shock Proteins

Listeria monocytogenes has evolved an extensive array of mechanisms for coping with stress and adapting to changing environmental conditions, ensuring its virulence phenotype expression. For this reason, L. monocytogenes has been identified as a significant food safety and public health concern. Among these adaptation systems are cold shock proteins (Csps), which facilitate rapid response to stress exposure. L. monocytogenes has three highly conserved csp genes, namely, cspA, cspB, and cspD. Using a series of csp deletion mutants, it has been shown that L. monocytogenes Csps are important for biofilm formation, motility, cold, osmotic, desiccation, and oxidative stress tolerance. Moreover, they are involved in overall virulence by impacting the expression of virulence-associated phenotypes, such as hemolysis and cell invasion. It is postulated that during stress exposure, Csps function to counteract harmful effects of stress, thereby preserving cell functions, such as DNA replication, transcription and translation, ensuring survival and growth of the cell. Interestingly, it seems that Csps might suppress tolerance to some stresses as their removal resulted in increased tolerance to stresses, such as desiccation for some strains. Differences in csp roles among strains from different genetic backgrounds are apparent for desiccation tolerance and biofilm production. Additionally, hierarchical trends for the different Csps and functional redundancies were observed on their influences on stress tolerance and virulence. Overall current data suggest that Csps have a wider role in bacteria physiology than previously assumed.

scholarly journals Localization of Cold Shock Proteins to Cytosolic Spaces Surrounding Nucleoids in Bacillus subtilis Depends on Active Transcription

2001 ◽  
Vol 183 (21) ◽  
pp. 6435-6443 ◽  
Author(s):  
Michael H. W. Weber ◽  
Arsen V. Volkov ◽  
Ingo Fricke ◽  
Mohamed A. Marahiel ◽  
Peter L. Graumann
Keyword(s):  
Bacillus Subtilis ◽  
Cold Shock ◽  
Fluorescent Protein ◽  
Cold Shock Protein ◽  
Initiation Of Translation ◽  
Specific Localization ◽  
Active Transcription ◽  
Green Fluorescent ◽  
Shock Proteins ◽  
Mutant Cells

ABSTRACT Using immunofluorescence microscopy and a fusion of a cold shock protein (CSP), CspB, to green fluorescent protein (GFP), we showed that in growing cells Bacillus subtilis CSPs specifically localize to cytosolic regions surrounding the nucleoid. The subcellular localization of CSPs is influenced by the structure of the nucleoid. Decondensed chromosomes in smc mutant cells reduced the sizes of the regions in which CSPs localized, while cold shock-induced chromosome compaction was accompanied by an expansion of the space in which CSPs were present. As a control, histone-like protein HBsu localized to the nucleoids, while β-galactosidase and GFP were detectable throughout the cell. After inhibition of translation, CspB-GFP was still present around the nucleoids in a manner similar to that in cold-shocked cells. However, in stationary-phase cells and after inhibition of transcription, CspB was distributed throughout the cell, indicating that specific localization of CspB depends on active transcription and is not due to simple exclusion from the nucleoid. Furthermore, we observed that nucleoids are more condensed and frequently abnormal incspB cspC and cspB cspDdouble-mutant cells. This suggests that the function of CSPs affects chromosome structure, probably through coupling of transcription to translation, which is thought to decondense nucleoids. In addition, we found that cspB cspD and cspB cspC double mutants are defective in sporulation, with a block at or before stage 0. Interestingly, CspB and CspC are depleted from the forespore compartment but not from the mother cell. In toto, our findings suggest that CSPs localize to zones of newly synthesized RNA, coupling transcription with initiation of translation.

Assessment of Sanitation Practices for the Control of Listeria monocytogenes at Small and Very Small Ready-To-Eat Meat and Poultry Processors

2021 ◽  
Author(s):  
Sheena E Fate ◽  
Jeannine P Schweihofer ◽  
Tina Conklin
Keyword(s):  
Listeria Monocytogenes ◽  
Microbial Control ◽  
Control Measures ◽  
Personal Hygiene ◽  
Control Methods ◽  
Department Of Agriculture ◽  
Food Contact Surfaces ◽  
Content Domain ◽  
Single Temperature ◽  
Pathogen Control

Science-based guidance employed at eight small and very small state and federally inspected ready-to-eat (RTE) meat and poultry processors across Michigan was assessed. Data was collected to determine the current level of sanitary control methods used for reducing Listeria in the processing environment and compared interactions with the facility microbial results. A checklist was created to assess the current recommended sanitary control methods from the U.S. Department of Agriculture; U.S. Food and Drug Administration; and the Michigan Department of Agriculture and Rural Development. The checklist, comprised of 178 items divided into ten general content domains, was used to assess which of the recommended controls were being utilized in the facilities to prevent post lethality contamination of RTE products . Effectiveness of pre-operational and operational sanitation was assessed through sampling 12 non-food contact surfaces using an adenosine triphosphate (ATP) reader and amplified nucleic single temperature reaction test for Listeria spp., including Listeria monocytogenes at each facility. In total, 288 samples were taken collectively from the 8 facilities (96 ATP, 96 pre-operational  Listeria  spp.). Microbial outcomes did not differ ( P > 0.05 ) based on the overall number of recommended sanitary control methods utilized and type of facility inspection. There was a greater content domain compliance overall in operational sanitation ( P  = 0.0005), sanitation ( P  = 0.0030), facility ( P  = 0.0397) and personal hygiene ( P = 0.0033 ) than for segregation procedures regardless of the regulating body. Findings suggest that regardless of the regulating body, the quality of sanitary control measures utilized is more impactful for microbial control than simply the quantity implemented. Pathogen control may be obtained without implementing all of the sanitary control methods within the guidance documents.

scholarly journals Restart of Exponential Growth of Cold-Shocked Yersinia enterocolitica Occurs after Down-Regulation ofcspA1/A2 mRNA

2000 ◽  
Vol 182 (11) ◽  
pp. 3285-3288 ◽  
Author(s):  
Klaus Neuhaus ◽  
Sonja Rapposch ◽  
Kevin P. Francis ◽  
Siegfried Scherer
Keyword(s):  
Yersinia Enterocolitica ◽  
Exponential Growth ◽  
Cold Shock ◽  
Cold Shock Protein ◽  
Clear Correlation ◽  
Wild Type ◽  
Tandem Gene Duplication ◽  
Cellular Content ◽  
Shock Proteins ◽  
Mutant Cells

ABSTRACT The cellular content of major cold shock protein (MCSP) mRNA transcribed from the tandem gene duplication cspA1/A2 and growth of Yersinia enterocolitica were compared when exponentially growing cultures of this bacterium were cold shocked from 30 to 20, 15, 10, 5, or 0°C, respectively. A clear correlation between the time point when exponential growth resumes after cold shock and the degradation of cspA1/A2 mRNA was found. A polynucleotide phosphorylase-deficient mutant was unable to degradecspA1/A2 mRNA properly and showed a delay, as well as a lower rate, of growth after cold shock. For this mutant, a correlation between decreasing cspA1/A2 mRNA and restart of growth after cold shock was also observed. For both wild-type and mutant cells, no correlation of restart of growth with the cellular content of MCSPs was found. We suggest that, after synthesis of cold shock proteins and cold adaptation of the cells, MCSP mRNAs must be degraded; otherwise, they trap ribosomes, prevent translation of bulk mRNA, and thus inhibit growth of this bacterium at low temperatures.

scholarly journals Biofilm Formation among Clinical and Food Isolates ofListeria monocytogenes

2013 ◽  
Vol 2013 ◽  
pp. 1-6 ◽  
Author(s):  
Joana Barbosa ◽  
Sandra Borges ◽  
Ruth Camilo ◽  
Rui Magalhães ◽  
Vânia Ferreira ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Listeria Monocytogenes ◽  
Osmotic Stress ◽  
Environmental Conditions ◽  
Biofilm Formation ◽  
Stress Conditions ◽  
Acidic Stress ◽  
Biofilm Production ◽  
Oflisteria Monocytogenes ◽  
Clinical Cases

Objective. A total of 725Listeria monocytogenesisolates, 607 from various foods and 118 from clinical cases of listeriosis, were investigated concerning their ability to form biofilms, at 4°C during 5 days and at 37°C during 24 h.Methods. Biofilm production was carried out on polystyrene tissue culture plates. FiveL. monocytogenesisolates were tested for biofilm formation after being exposed to acidic and osmotic stress conditions.Results. Significant differences (P<0.01) between clinical and food isolates were observed. At 37°C for 24 h, most food isolates were classified as weak or moderate biofilm formers whereas all the clinical isolates were biofilm producers, although the majority were weak. At 4°C during 5 days, 65 and 59% isolates, from food and clinical cases, respectively, were classified as weak. After both sublethal stresses, at 37°C just one of the five isolates tested was shown to be more sensitive to subsequent acidic exposure. However, at 4°C both stresses did not confer either sensitivity or resistance.Conclusions. Significant differences between isolates origin, temperature, and sublethal acidic stress were observed concerning the ability to form biofilms. Strain, origin, and environmental conditions can determine the level of biofilm production byL. monocytogenesisolates.

scholarly journals Role of ctc from Listeria monocytogenes in Osmotolerance

2003 ◽  
Vol 69 (1) ◽  
pp. 154-161 ◽  
Author(s):  
Rozenn Gardan ◽  
Ophélie Duché ◽  
Sabine Leroy-Sétrin ◽  
Jean Labadie
Keyword(s):  
Salt Stress ◽  
Heat Stress ◽  
Listeria Monocytogenes ◽  
Osmotic Stress ◽  
Minimal Medium ◽  
Stress Protein ◽  
Transcriptional Level ◽  
Osmotic Stress Tolerance ◽  
Electron Microscopy Analysis

ABSTRACT Listeria monocytogenes is a food-borne pathogen with the ability to grow under conditions of high osmolarity. In a previous study, we reported the identification of 12 proteins showing high induction after salt stress. One of these proteins is highly similar to the general stress protein Ctc of Bacillus subtilis. In this study, induction of Ctc after salt stress was confirmed at the transcriptional level by using RNA slot blot experiments. To explore the role of the ctc gene product in resistance to stresses, we constructed a ctc insertional mutant. No difference in growth was observed between the wild-type strain LO28 and the ctc mutant either in rich medium after osmotic or heat stress or in minimal medium after heat stress. However, in minimal medium after osmotic stress, the growth rate of the mutant was increased by a factor of 2. Moreover, electron microscopy analysis showed impaired morphology of the mutant grown under osmotic stress conditions in minimal medium. Addition of the osmoprotectant glycine betaine to the medium completely abolished the osmotic sensitivity phenotype of the ctc mutant. Altogether, these results suggest that the Ctc protein of L. monocytogenes is involved in osmotic stress tolerance in the absence of any osmoprotectant in the medium.

scholarly journals Selective mRNA Degradation by Polynucleotide Phosphorylase in Cold Shock Adaptation in Escherichia coli

2001 ◽  
Vol 183 (9) ◽  
pp. 2808-2816 ◽  
Author(s):  
Kunitoshi Yamanaka ◽  
Masayori Inouye
Keyword(s):  
Escherichia Coli ◽  
Cell Growth ◽  
Cold Shock ◽  
Critical Role ◽  
Rna Helicase ◽  
Cold Shock Protein ◽  
Polynucleotide Phosphorylase ◽  
Selective Degradation ◽  
High Level ◽  
Shock Proteins

ABSTRACT Upon cold shock, Escherichia coli cell growth transiently stops. During this acclimation phase, specific cold shock proteins (CSPs) are highly induced. At the end of the acclimation phase, their synthesis is reduced to new basal levels, while the non-cold shock protein synthesis is resumed, resulting in cell growth reinitiation. Here, we report that polynucleotide phosphorylase (PNPase) is required to repress CSP production at the end of the acclimation phase. A pnp mutant, upon cold shock, maintained a high level of CSPs even after 24 h. PNPase was found to be essential for selective degradation of CSP mRNAs at 15°C. In a poly(A) polymerase mutant and a CsdA RNA helicase mutant, CSP expression upon cold shock was significantly prolonged, indicating that PNPase in concert with poly(A) polymerase and CsdA RNA helicase plays a critical role in cold shock adaptation.

scholarly journals Three Transporters Mediate Uptake of Glycine Betaine and Carnitine by Listeria monocytogenes in Response to Hyperosmotic Stress

2003 ◽  
Vol 69 (2) ◽  
pp. 1013-1022 ◽  
Author(s):  
Apostolos S. Angelidis ◽  
Gary M. Smith
Keyword(s):  
Listeria Monocytogenes ◽  
Osmotic Stress ◽  
Glycine Betaine ◽  
Nacl Stress ◽  
Food Preservation ◽  
Transport Systems ◽  
Triple Mutant ◽  
Carnitine Transporter ◽  
Food Borne ◽  
Carnitine Transport

ABSTRACT The uptake and accumulation of the potent osmolytes glycine betaine and carnitine enable the food-borne pathogen Listeria monocytogenes to proliferate in environments of elevated osmotic stress, often rendering salt-based food preservation inadequate. To date, three osmolyte transport systems are known to operate in L. monocytogenes: glycine betaine porter I (BetL), glycine betaine porter II (Gbu), and a carnitine transporter OpuC. We investigated the specificity of each transporter towards each osmolyte by creating mutant derivatives of L. monocytogenes 10403S that possess each of the transporters in isolation. Kinetic and steady-state osmolyte accumulation data together with growth rate experiments demonstrated that osmotically activated glycine betaine transport is readily and effectively mediated by Gbu and BetL and to a lesser extent by OpuC. Osmotically stimulated carnitine transport was demonstrated for OpuC and Gbu regardless of the nature of stressing salt. BetL can mediate weak carnitine uptake in response to NaCl stress but not KCl stress. No other transporter in L. monocytogenes 10403S appears to be involved in osmotically stimulated transport of either osmolyte, since a triple mutant strain yielded neither transport nor accumulation of glycine betaine or carnitine and could not be rescued by either osmolyte when grown under elevated osmotic stress.

scholarly journals Pepper Novel Pseudo Response Regulator Protein CaPRR2 Modulates Drought and High Salt Tolerance

2021 ◽  
Vol 12 ◽  
Author(s):  
Junsub Lim ◽  
Chae Woo Lim ◽  
Sung Chul Lee
Keyword(s):  
Osmotic Stress ◽  
Stress Tolerance ◽  
Response Regulator ◽  
Environmental Stresses ◽  
Stress Conditions ◽  
Osmotic Stress Tolerance ◽  
Regulator Protein ◽  
Pseudo Response Regulator ◽  
Response Regulator Protein ◽  
Pepper Plants

Plants modify their internal states to adapt to environmental stresses. Under environmental stress conditions, plants restrict their growth and development and activate defense responses. Abscisic acid (ABA) is a major phytohormone that plays a crucial role in the osmotic stress response. In osmotic stress adaptation, plants regulate stomatal closure, osmoprotectant production, and gene expression. Here, we isolated CaPRR2 – encoding a pseudo response regulator protein – from the leaves of pepper plants (Capsicum annuum). After exposure to ABA and environmental stresses, such as drought and salt stresses, CaPRR2 expression in pepper leaves was significantly altered. Under drought and salt stress conditions, CaPRR2-silenced pepper plants exhibited enhanced osmotic stress tolerance, characterized by an enhanced ABA-induced stomatal closing and high MDA and proline contents, compared to the control pepper plants. Taken together, our data indicate that CaPRR2 negatively regulates osmotic stress tolerance.

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