Genomic and transcriptomic analyses reveal distinct biological functions for cold shock proteins (VpaCspA and VpaCspD) in Vibrio parahaemolyticus CHN25 during low-temperature survival

ABSTRACT Members of the group of 7-kDa cold-shock proteins (CSPs) are the proteins with the highest level of induction upon cold shock in the lactic acid bacterium Lactococcus lactis MG1363. By using double-crossover recombination, two L. lactis strains were generated in which genes encoding CSPs are disrupted: L. lactis NZ9000ΔAB lacks the tandemly orientatedcspA and cspB genes, and NZ9000ΔABE lackscspA, cspB, and cspE. Both strains showed no differences in growth at normal and at low temperatures compared to that of the wild-type strain, L. lactis NZ9000. Two-dimensional gel electrophoresis showed that upon disruption of thecspAB genes, the production of remaining CspE at low temperature increased, and upon disruption of cspA, cspB, and cspE, the production of CspD at normal growth temperatures increased. Northern blot analysis showed that control is most likely at the transcriptional level. Furthermore, it was established by a proteomics approach that some (non-7-kDa) cold-induced proteins (CIPs) are not cold induced in the csp-lacking strains, among others the histon-like protein HslA and the signal transduction protein LlrC. This supports earlier observations (J. A. Wouters, M. Mailhes, F. M. Rombouts, W. M. De Vos, O. P. Kuipers, and T. Abee, Appl. Environ. Microbiol. 66:3756–3763, 2000). that the CSPs of L. lactis might be directly involved in the production of some CIPs upon low-temperature exposure. Remarkably, the adaptive response to freezing by prior exposure to 10°C was significantly reduced in strain NZ9000ΔABE but not in strain NZ9000ΔAB compared to results with wild-type strain NZ9000, indicating a notable involvement of CspE in cryoprotection.

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Cold Shock Proteins and Low-Temperature Response ofStreptococcus thermophilus CNRZ302

Applied and Environmental Microbiology ◽

10.1128/aem.65.10.4436-4442.1999 ◽

1999 ◽

Vol 65 (10) ◽

pp. 4436-4442 ◽

Cited By ~ 39

Author(s):

Jeroen A. Wouters ◽

Frank M. Rombouts ◽

Willem M. de Vos ◽

Oscar P. Kuipers ◽

Tjakko Abee

Keyword(s):

Low Temperature ◽

Cold Shock ◽

Thermophilic Bacterium ◽

Temperature Adaptation ◽

Transcriptional Level ◽

Minimal Growth ◽

Two Dimensional Gel Electrophoresis ◽

Cold Shock Proteins ◽

Shock Proteins ◽

Induced Proteins

ABSTRACT Low-temperature adaptation and cryoprotection were studied in the thermophilic lactic acid bacterium Streptococcus thermophilus CNRZ302. S. thermophilus actively adapts to freezing during a pretreatment at 20°C, resulting in an approximately 1,000-fold increased survival after four freeze-thaw cycles compared to mid-exponential-phase cells grown at an optimal temperature of 42°C. No adaptation is observed when cells are exposed to a temperature (10°C) below the minimal growth temperature of the strain (just below 15°C). By two-dimensional gel electrophoresis several 7-kDa cold-induced proteins were identified, which are the major induced proteins after a shift to 20°C. These cold shock proteins were maximally expressed at 20°C, while the induction level was low after cold shock to 10°C. To confirm the presence ofcsp genes in S. thermophilus, a PCR strategy was used which yielded products of different sizes. Sequence analysis revealed csp-like sequences that were up to 95% identical to those of csp genes of S. thermophilus ST1-1,Streptococcus dysgalactiae, Streptococcus pyogenes, and Lactococcus lactis. Northern blot analysis revealed a seven- to ninefold induction of cspmRNA after a temperature shift to 20°C, showing that this thermophilic bacterium indeed contains at least one cold-induciblecsp gene and that its regulation takes place at the transcriptional level.

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The Role of Cold-Shock Proteins in Low-Temperature Adaptation of Food-Related Bacteria

Systematic and Applied Microbiology ◽

10.1016/s0723-2020(00)80001-6 ◽

2000 ◽

Vol 23 (2) ◽

pp. 165-173 ◽

Cited By ~ 50

Author(s):

Jeroen A. Wouters ◽

Frank M. Rombouts ◽

Oscar P. Kuipers ◽

Willem M. de Vos ◽

T. Abee

Keyword(s):

Low Temperature ◽

Cold Shock ◽

Temperature Adaptation ◽

Cold Shock Proteins ◽

Shock Proteins ◽

Low Temperature Adaptation

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Enhanced Levels of Cold Shock Proteins in Listeria monocytogenes LO28 upon Exposure to Low Temperature and High Hydrostatic Pressure

Applied and Environmental Microbiology ◽

10.1128/aem.68.2.456-463.2002 ◽

2002 ◽

Vol 68 (2) ◽

pp. 456-463 ◽

Cited By ~ 90

Author(s):

Henrike H. Wemekamp-Kamphuis ◽

Andreas K. Karatzas ◽

Jeroen A. Wouters ◽

Tjakko Abee

Keyword(s):

Listeria Monocytogenes ◽

Hydrostatic Pressure ◽

Low Temperature ◽

High Hydrostatic Pressure ◽

Cold Shock ◽

Chromosomal Dna ◽

Two Dimensional Gel Electrophoresis ◽

Cold Shock Proteins ◽

Food Borne ◽

Shock Proteins

ABSTRACT Listeria monocytogenes is a psychrotrophic food-borne pathogen that is problematic for the food industry because of its ubiquitous distribution in nature and its ability to grow at low temperatures and in the presence of high salt concentrations. Here we demonstrate that the process of adaptation to low temperature after cold shock includes elevated levels of cold shock proteins (CSPs) and that the levels of CSPs are also elevated after treatment with high hydrostatic pressure (HHP). Two-dimensional gel electrophoresis combined with Western blotting performed with anti-CspB of Bacillus subtilis was used to identify four 7-kDa proteins, designated Csp1, Csp2, Csp3, and Csp4. In addition, Southern blotting revealed four chromosomal DNA fragments that reacted with a csp probe, which also indicated that a CSP family is present in L. monocytogenes LO28. After a cold shock in which the temperature was decreased from 37Ã¯Â¿Â½C to 10Ã¯Â¿Â½C the levels of Csp1 and Csp3 increased 10- and 3.5-fold, respectively, but the levels of Csp2 and Csp4 were not elevated. Pressurization of L. monocytogenes LO28 cells resulted in 3.5- and 2-fold increases in the levels of Csp1 and Csp2, respectively. Strikingly, the level of survival after pressurization of cold-shocked cells was 100-fold higher than that of cells growing exponentially at 37Ã¯Â¿Â½C. These findings imply that cold-shocked cells are protected from HHP treatment, which may affect the efficiency of combined preservation techniques.

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Adaptation of anammox bacteria to low temperature via gradual acclimation and cold shocks: distinctions in protein expression, membrane composition and activities

10.1101/2021.08.29.458117 ◽

2021 ◽

Author(s):

V. Kouba ◽

D. Vejmelkova ◽

E. Zwolsman ◽

K. Hurkova ◽

K. Navratilova ◽

...

Keyword(s):

Low Temperature ◽

Protein Expression ◽

Low Temperatures ◽

Cold Shock ◽

Anammox Bacteria ◽

List Type ◽

Temperature Decrease ◽

Cold Shock Proteins ◽

Shock Proteins ◽

Ladderane Lipids

AbstractAnammox bacteria enable an efficient removal of nitrogen from sewage in processes involving partial nitritation and anammox (PN/A) or nitrification, partial denitrification, and anammox (N-PdN/A). In mild climates, anammox bacteria must be adapted to ≤15 °C, typically by gradual temperature decrease; however, this takes months or years. To reduce the time necessary for the adaptation, an unconventional method of ‘cold shocks’ is promising, involving hours-long exposure of anammox biomass to extremely low temperatures. We compared the efficacies of gradual temperature decrease and cold shocks to increase the metabolic activity of anammox (fed batch reactor, planktonic “Ca. Kuenenia”). We assessed the cold shock mechanism on the level of protein expression (quantitative shot-gun proteomics, LC-HRMS/MS) and structure of membrane lipids (UPLC-HRMS/MS). The shocked culture was more active (0.66±0.06 vs 0.48±0.06 kg-N/kg-VSS/d) and maintained the relative content of N-respiration proteins at levels consistent levels with the initial state, whereas the content of these proteins decreased in gradually acclimated culture. Cold shocks also induced a more efficient up-regulation of cold shock proteins (e.g. CspB, TypA, ppiD). Ladderane lipids characteristic for anammox evolved to a similar end-point in both cultures which confirms their role in anammox bacteria adaptation to cold and indicates a three-pronged adaptation mechanism involving ladderane lipids (ladderane alkyl length, introduction of shorter non-ladderane alkyls, polar headgroup). Overall, we show the outstanding potential of cold shocks for low-temperature adaptation of anammox bacteria and provide yet unreported detailed mechanisms of anammox adaptation to low temperatures.HighlightsAnammox bacteria were adapted to low T by gradual acclimation and cold shocksThe shocked culture was more active (0.66±0.06 vs 0.48±0.06 kg-N/kg-VSS/d)N-respiration proteins content decreased in gradually acclimated bacteriaSeveral cold shock proteins were upregulated more efficiently by cold shocksAt ↓T, anammox adjusted ladderane membrane lipid composition in three aspectsGraphical abstract

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Chapter 4 The role of cold-shock proteins in low-temperature adaptation

Cell and Molecular Response to Stress - Protein Adaptations and Signal Transduction ◽

10.1016/s1568-1254(01)80006-1 ◽

2001 ◽

pp. 43-56 ◽

Cited By ~ 1

Author(s):

Jeroen A. Wouters ◽

Frank M. Rombouts ◽

Oscar P. Kuipers ◽

Willem M. de Vos ◽

T. Abee

Keyword(s):

Low Temperature ◽

Cold Shock ◽

Temperature Adaptation ◽

Cold Shock Proteins ◽

Shock Proteins ◽

Low Temperature Adaptation

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

Microorganisms ◽

10.3390/microorganisms9051061 ◽

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.

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