Analyzing possible native functions of the quinolone resistance gene qnr in Vibrio vulnificus

The worldwide distribution of qnr genes found on plasmids and their presence on the chromosomes of aquatic bacteria, like Vibrio vulnificus, one of the suspected sources, suggests an origin before the development of synthetic quinolones. However, their native function remains unknown. Previous work indicated that expression of qnrVv in V. vulnificus was induced by cold shock. To investigate its role further we constructed single in-frame deletion mutants in qnrVv and cspA (the gene for cold shock protein), and a double mutant in qnrVv and cspA in V. vulnificus ATCC 17562 to evaluate the response to different environmental conditions and stresses and to exposure to various DNA damaging agents. We found that qnrVv is involved in resistance to ciprofloxacin, levofloxacin, and mitomycin C, and in the cold shock response in V. vulnificus. Moreover, ΔqnrVv and ΔcspA mutants showed a lower growth when they were treated with bile salts at 37°C and then shifted to 15°C (cold shock) without bile salts in the media, with the effect being higher in the double mutant. This transition may mimic what happens when V. vulnificus is ingested into the gastrointestinal tract and released in its natural environment. Cold shock and bile salts induced the expression of cspA and DNA gyrase and topoisomerase IV genes. However, no induction was found in the ΔqnrVv mutant, suggesting that the qnrVv gene is involved in the response to DNA damage and nucleic acid secondary structure.

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RbfA, a 30S ribosomal binding factor, is a cold-shock protein whose absence triggers the cold-shock response

Molecular Microbiology ◽

10.1111/j.1365-2958.1996.tb02582.x ◽

1996 ◽

Vol 21 (6) ◽

pp. 1207-1218 ◽

Cited By ~ 131

Author(s):

Pamela G. Jones ◽

Masayori Inouye

Keyword(s):

Cold Shock ◽

Cold Shock Protein ◽

Cold Shock Response ◽

Binding Factor ◽

Shock Response

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Transcriptomic time-series analysis of cold- and heat-shock response in psychrotrophic lactic acid bacteria

10.21203/rs.3.rs-34513/v1 ◽

2020 ◽

Author(s):

Ilhan Cem Duru ◽

Anne Ylinen ◽

Sergei Belanov ◽

Alan Ávila Pulido ◽

Lars Paulin ◽

...

Keyword(s):

Lactic Acid ◽

Heat Shock ◽

Lactic Acid Bacteria ◽

Heat Shock Response ◽

Network Inference ◽

Cold Shock ◽

Cold Shock Protein ◽

Gene Network Inference ◽

Cold Shock Response ◽

Shock Response

Abstract Background: Psychrotrophic lactic acid bacteria (LAB) species are the dominant species in microbiota of cold-stored modified-atmosphere-packaged food products and they are the main cause of food spoilage. But still, the cold- and heat-shock response of the spoilage-related psychrotrophic lactic acid bacteria has not been studied. Here, to study cold- and heat-shock response of spoilage lactic acid bacteria, we performed time-series RNA-seq for Le. gelidum, Lc. piscium and P. oligofermentans using temperatures of 0 °C, 4 °C, 14 °C, 25 °C and 28 °C. Results: We showed that the cold-shock protein A (cspA) gene was the main cold-shock protein gene among cold-shock protein genes in all three species. Our results indicated DEAD-box RNA helicase genes (cshA, cshB) play a critical role in cold-shock response in psychrotrophic LAB. In addition, several RNase genes were also involved in cold-shock response in Lc. piscium and P. oligofermentans. Moreover, gene network inference analysis provided candidate genes involved in cold-shock response. Ribosomal proteins, tRNA modification, rRNA modification, and ABC and efflux MFS transporter genes clustered with cold-shock response genes in all three species, which was a strong indication that these genes would be part of cold-shock response machinery. Heat-shock treatment caused upregulation of Clp protease and chaperone genes in all three species and we were able to identify transcription binding site motifs for heat-shock response genes in Le. gelidum and Lc. piscium. Finally, we showed that food spoilage-related genes were upregulated at cold temperatures. Conclusions: The results of this study provide new insights into a better understanding of the cold- and heat-shock response in psychrotrophic LAB. In addition, candidate genes involved in cold- and heat-shock response predicted using gene network inference analysis could be used as a target for future studies.

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CspB and CspC are induced upon cold shock in Bacillus cereus strain D2

Canadian Journal of Microbiology ◽

10.1139/cjm-2021-0025 ◽

2021 ◽

Author(s):

Haoyang Li ◽

Rui Yang ◽

Linlin Hao ◽

Chunli Wang ◽

Mingtang Li

Keyword(s):

Bacillus Cereus ◽

Contaminated Soils ◽

Cold Shock ◽

Amino Acid Sequences ◽

Cold Shock Protein ◽

E Coli ◽

Subsequent Transformation ◽

Cold Shock Response ◽

Shock Response ◽

Psychrotrophic Strain

Bacillus cereus D2, a psychrotrophic strain, plays an essential role in the restoration of heavy metal-contaminated soils, especially at low temperatures. However, the cold shock response mechanisms of this strain are unclear. In this study, the cold shock response of B. cereus D2 was characterized; as per the Arrhenius curve, 10 °C was chosen as the cold shock temperature. Six cold shock-like proteins were found and temporarily named cold shock protein (Csp)1-6; the respective genes were cloned and identified. Quantitative real-time PCR results showed that csp1, csp2, csp3, and csp6 were overexpressed under cold shock conditions. Interestingly, after cloning the respective encoding genes into pET-28a (+) vector and their subsequent transformation into E. coli BL21 (DE3), the strains expressing Csp2 and Csp6 grew faster at 10 °C, showing a large number of bacteria. These results suggest that Csp2 and Csp6 are the major cold shock proteins in B. cereus D2. Of note, the comparison of amino acid sequences and structures showed that Csp2 and Csp6 belong to the CspB and CspC families, respectively. Additionally, we show that the number of hydrophobic residues is not a determining feature of major Csps, while, on the other hand, the formation of an α-helix in the context of a leucine residue is the most dominant difference between major, and other Bacillus and E. coli Csps.

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Cold Temperature Adaptation and Growth of Microorganisms†

Journal of Food Protection ◽

10.4315/0362-028x-60.12.1583 ◽

1997 ◽

Vol 60 (12) ◽

pp. 1583-1594 ◽

Cited By ~ 85

Author(s):

ELAINE D. BERRY ◽

PEGGY M. FOEGEDING

Keyword(s):

Cold Shock ◽

Cold Temperature ◽

Temperature Adaptation ◽

Cold Shock Protein ◽

Conformational Structure ◽

Temperature Growth ◽

Psychrotrophic Bacteria ◽

Cold Shock Response ◽

Shock Response ◽

Cold Tolerant

Most microorganisms must accommodate a variety of changing conditions and stresses in their environment in order to survive and multiply. Because of the impact of temperature on all reactions of the cell, adaptations to fluctuations in temperature are possibly the most common. Widespread in the environment and well-equipped for cold temperature growth, psychrophilic and psychrotrophic microorganisms may yet make numerous adjustments when faced with temperatures lower than optimum. Phospholipid and fatty acid alterations resulting in increased membrane fluidity at lower temperatures have been described for many cold tolerant microorganisms while others may make no similar adjustment. While the enzymes of cold growing bacteria have been less extensively studied than those of thermophilic bacteria, it appears that function at low temperature requires enzymes with flexible conformational structure, in order to compensate for lower reaction rates. In many organisms, including psychrophilic and psychrotrophic bacteria, specific sets of cold shock proteins are induced upon abrupt shifts to colder temperatures. While this cold shock response has not been fully delineated, it appears to be adaptive, and may function to promote the expression of genes involved in translation when cells are displaced to lower temperatures. The cold shock response of Escherichia coli has been extensively studied, and the major cold shock protein CspA appears to be involved in the regulation of the response. Upon cold shock, the induction of CspA and its counterparts in most microorganisms studied is prominent, but transient; studies of this response in some psychrotrophic bacteria have reported constitutive synthesis and continued synthesis during cold temperature growth of CspA homologues, and it will be interesting to learn if these are common mechanisms of among cold tolerant organisms. Psychrotrophic microorganisms continue to be a spoilage and safety problem in refrigerated foods, and a greater understanding of the physiological mechanisms and implications of cold temperature adaptation and growth should enhance our ability to design more effective methods of preservation.

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A comparison of the chilling-stress response in two differentially tolerant cultivars of tomato (Lycopersicon esculentum)

Biochemistry and Cell Biology ◽

10.1139/o92-029 ◽

1992 ◽

Vol 70 (3-4) ◽

pp. 191-198 ◽

Cited By ~ 1

Author(s):

Randal W. Giroux ◽

W. Gary Filion

Keyword(s):

Cold Shock ◽

Chilling Stress ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Protein S ◽

Cold Shock Protein ◽

Relative Mass ◽

Temperature Decrease ◽

Cold Shock Response

The chilling responses of two differentially cold tolerant cultivars of tomato were monitored through in vivo labelling of polypeptides with [35S]methionine, both during a gradual temperature decrease (2 °C/day) and also during a rapid cold shock (4 °C). The polypeptides were separated by one-dimensional sodium dodecyl sulfate – polyacrylamide gel electrophoresis and revealed by fluorography. Both cultivars showed changes in the polypeptide profiles resulting from either chilling treatment. During the gradual temperature decrease, there were few differences exhibited between the two cultivars. However, during cold shock both cultivars showed the altered synthesis of several unique polypeptides. Both cultivars showed the appearance of a 35-kDa polypeptide during the gradual temperature decrease and also during the cold shock. The appearance of three high relative mass polypeptides was found in both cultivars only during the gradual temperature decrease. Treatments with cycloheximide and chloramphenicol suggested that cold-shock polypeptides are both nuclear and organelle encoded. The cold-shock response in roots was different from the response in leaves and between cultivars. A comparison of the two cultivars showed a number of differences in polypeptide synthesis which may be related to increased cold tolerance.Key words: cold-shock protein(s), tomato, chilling stress, acclimation, cold tolerance.

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RNA-Seq-Based Analysis of Cold Shock Response in Thermoanaerobacter tengcongensis, a Bacterium Harboring a Single Cold Shock Protein Encoding Gene

PLoS ONE ◽

10.1371/journal.pone.0093289 ◽

2014 ◽

Vol 9 (3) ◽

pp. e93289 ◽

Cited By ~ 13

Author(s):

Bo Liu ◽

Yuhong Zhang ◽

Wei Zhang

Keyword(s):

Cold Shock ◽

Cold Shock Protein ◽

Rna Seq ◽

Cold Shock Response ◽

Thermoanaerobacter Tengcongensis ◽

Protein Encoding ◽

Shock Response ◽

Encoding Gene

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Transcriptomic time-series analysis of cold- and heat-shock response in psychrotrophic lactic acid bacteria

BMC Genomics ◽

10.1186/s12864-020-07338-8 ◽

2021 ◽

Vol 22 (1) ◽

Author(s):

Ilhan Cem Duru ◽

Anne Ylinen ◽

Sergei Belanov ◽

Alan Avila Pulido ◽

Lars Paulin ◽

...

Keyword(s):

Lactic Acid ◽

Heat Shock ◽

Lactic Acid Bacteria ◽

Heat Shock Response ◽

Network Inference ◽

Cold Shock ◽

Cold Shock Protein ◽

Gene Network Inference ◽

Cold Shock Response ◽

Shock Response

Abstract Background Psychrotrophic lactic acid bacteria (LAB) species are the dominant species in the microbiota of cold-stored modified-atmosphere-packaged food products and are the main cause of food spoilage. Despite the importance of psychrotrophic LAB, their response to cold or heat has not been studied. Here, we studied the transcriptome-level cold- and heat-shock response of spoilage lactic acid bacteria with time-series RNA-seq for Le. gelidum, Lc. piscium, and P. oligofermentans at 0 °C, 4 °C, 14 °C, 25 °C, and 28 °C. Results We observed that the cold-shock protein A (cspA) gene was the main cold-shock protein gene in all three species. Our results indicated that DEAD-box RNA helicase genes (cshA, cshB) also play a critical role in cold-shock response in psychrotrophic LAB. In addition, several RNase genes were involved in cold-shock response in Lc. piscium and P. oligofermentans. Moreover, gene network inference analysis provided candidate genes involved in cold-shock response. Ribosomal proteins, tRNA modification, rRNA modification, and ABC and efflux MFS transporter genes clustered with cold-shock response genes in all three species, indicating that these genes could be part of the cold-shock response machinery. Heat-shock treatment caused upregulation of Clp protease and chaperone genes in all three species. We identified transcription binding site motifs for heat-shock response genes in Le. gelidum and Lc. piscium. Finally, we showed that food spoilage-related genes were upregulated at cold temperatures. Conclusions The results of this study provide new insights on the cold- and heat-shock response of psychrotrophic LAB. In addition, candidate genes involved in cold- and heat-shock response predicted using gene network inference analysis could be used as targets for future studies.

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