scholarly journals Heat Shock Protein-Mediated Resistance to High Hydrostatic Pressure in Escherichia coli

2004 ◽  
Vol 70 (5) ◽  
pp. 2660-2666 ◽  
Author(s):  
Abram Aertsen ◽  
Kristof Vanoirbeek ◽  
Philipp De Spiegeleer ◽  
Jan Sermon ◽  
Kristel Hauben ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
High Pressure ◽  
Hydrostatic Pressure ◽  
Heat Shock ◽  
Heat Shock Proteins ◽  
High Hydrostatic Pressure ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
E Coli ◽  
Shock Proteins

ABSTRACT A random library of Escherichia coli MG1655 genomic fragments fused to a promoterless green fluorescent protein (GFP) gene was constructed and screened by differential fluorescence induction for promoters that are induced after exposure to a sublethal high hydrostatic pressure stress. This screening yielded three promoters of genes belonging to the heat shock regulon (dnaK, lon, clpPX), suggesting a role for heat shock proteins in protection against, and/or repair of, damage caused by high pressure. Several further observations provide additional support for this hypothesis: (i) the expression of rpoH, encoding the heat shock-specific sigma factor σ32, was also induced by high pressure; (ii) heat shock rendered E. coli significantly more resistant to subsequent high-pressure inactivation, and this heat shock-induced pressure resistance followed the same time course as the induction of heat shock genes; (iii) basal expression levels of GFP from heat shock promoters, and expression of several heat shock proteins as determined by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis of proteins extracted from pulse-labeled cells, was increased in three previously isolated pressure-resistant mutants of E. coli compared to wild-type levels.

scholarly journals Functional Analysis of Genes Comprising the Locus of Heat Resistance in Escherichia coli

2017 ◽  
Vol 83 (20) ◽  
Author(s):  
Ryan Mercer ◽  
Oanh Nguyen ◽  
Qixing Ou ◽  
Lynn McMullen ◽  
Michael G. Gänzle
Keyword(s):  
Escherichia Coli ◽  
Heat Shock ◽  
Heat Shock Proteins ◽  
Heat Resistance ◽  
Growth Phase ◽  
Genomic Island ◽  
Enteric Bacteria ◽  
Content Type ◽  
E Coli ◽  
Shock Proteins

ABSTRACT The locus of heat resistance (LHR) is a 15- to 19-kb genomic island conferring exceptional heat resistance to organisms in the family Enterobacteriaceae, including pathogenic strains of Salmonella enterica and Escherichia coli. The complement of LHR-comprising genes that is necessary for heat resistance and the stress-induced or growth-phase-induced expression of LHR-comprising genes are unknown. This study determined the contribution of the seven LHR-comprising genes yfdX1 GI, yfdX2, hdeD GI, orf11, trx GI, kefB, and psiE GI by comparing the heat resistances of E. coli strains harboring plasmid-encoded derivatives of the different LHRs in these genes. (Genes carry a subscript “GI” [genomic island] if an ortholog of the same gene is present in genomes of E. coli.) LHR-encoded heat shock proteins sHSP20, ClpKGI, and sHSPGI are not sufficient for the heat resistance phenotype; YfdX1, YfdX2, and HdeD are necessary to complement the LHR heat shock proteins and to impart a high level of resistance. Deletion of trx GI, kefB, and psiE GI from plasmid-encoded copies of the LHR did not significantly affect heat resistance. The effect of the growth phase and the NaCl concentration on expression from the putative LHR promoter p2 was determined by quantitative reverse transcription-PCR and by a plasmid-encoded p2:GFP promoter fusion. The expression levels of exponential- and stationary-phase E. coli cells were not significantly different, but the addition of 1% NaCl significantly increased LHR expression. Remarkably, LHR expression in E. coli was dependent on a chromosomal copy of evgA. In conclusion, this study improved our understanding of the genes required for exceptional heat resistance in E. coli and factors that increase their expression in food. IMPORTANCE The locus of heat resistance (LHR) is a genomic island conferring exceptional heat resistance to several foodborne pathogens. The exceptional level of heat resistance provided by the LHR questions the control of pathogens by current food processing and preparation techniques. The function of LHR-comprising genes and their regulation, however, remain largely unknown. This study defines a core complement of LHR-encoded proteins that are necessary for heat resistance and demonstrates that regulation of the LHR in E. coli requires a chromosomal copy of the gene encoding EvgA. This study provides insight into the function of a transmissible genomic island that allows otherwise heat-sensitive enteric bacteria, including pathogens, to lead a thermoduric lifestyle and thus contributes to the detection and control of heat-resistant enteric bacteria in food.

scholarly journals Inactivation of Escherichia coli andListeria innocua in Milk by Combined Treatment with High Hydrostatic Pressure and the Lactoperoxidase System

2000 ◽  
Vol 66 (10) ◽  
pp. 4173-4179 ◽  
Author(s):  
Cristina García-Graells ◽  
Caroline Valckx ◽  
Chris W. Michiels
Keyword(s):  
Escherichia Coli ◽  
High Pressure ◽  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Combined Treatment ◽  
Food Preservation ◽  
Pressure Treatment ◽  
E Coli ◽  
Preservation Method ◽  
Lactoperoxidase System

ABSTRACT We have studied inactivation of four strains each ofEscherichia coli and Listeria innocua in milk by the combined use of high hydrostatic pressure and the lactoperoxidase-thiocyanate-hydrogen peroxide system as a potential mild food preservation method. The lactoperoxidase system alone exerted a bacteriostatic effect on both species for at least 24 h at room temperature, but none of the strains was inactivated. Upon high-pressure treatment in the presence of the lactoperoxidase system, different results were obtained for E. coli and L. innocua. For none of the E. coli strains did the lactoperoxidase system increase the inactivation compared to a treatment with high pressure alone. However, a strong synergistic interaction of both treatments was observed for L. innocua. Inactivation exceeding 7 decades was achieved for all strains with a mild treatment (400 MPa, 15 min, 20°C), which in the absence of the lactoperoxidase system caused only 2 to 5 decades of inactivation depending on the strain. Milk as a substrate was found to have a considerable effect protecting E. coli and L. innocua against pressure inactivation and reducing the effectiveness of the lactoperoxidase system under pressure on L. innocua. Time course experiments showed that L. innocua counts continued to decrease in the first hours after pressure treatment in the presence of the lactoperoxidase system.E. coli counts remained constant for at least 24 h, except after treatment at the highest pressure level (600 MPa, 15 min, 20°C), in which case, in the presence of the lactoperoxidase system, a transient decrease was observed, indicating sublethal injury rather than true inactivation.

HIGH HYDROSTATIC PRESSURE INDUCES SYNTHESIS OF HEAT-SHOCK PROTEINS AND TREHALOSE-6-PHOSPHATE SYNTHASE INAnastrepha ludensLARVAE

2013 ◽  
Vol 82 (4) ◽  
pp. 196-212 ◽  
Author(s):  
Manuel A. Vargas-Ortiz ◽  
Rodolfo Quintana-Castro ◽  
Rosa M. Oliart-Ros ◽  
Javier De la Cruz-Medina ◽  
José A. Ramírez de León ◽  
...  
Keyword(s):  
Hydrostatic Pressure ◽  
Heat Shock ◽  
Heat Shock Proteins ◽  
High Hydrostatic Pressure ◽  
Shock Proteins ◽  
Phosphate Synthase

scholarly journals Proteomic and Transcriptomic Analysis of Microviridae φX174 Infection Reveals Broad Upregulation of Host Escherichia coli Membrane Damage and Heat Shock Responses

mSystems ◽  
2021 ◽  
Vol 6 (3) ◽  
Author(s):  
Bradley W. Wright ◽  
Dominic Y. Logel ◽  
Mehdi Mirzai ◽  
Dana Pascovici ◽  
Mark P. Molloy ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Heat Shock ◽  
Heat Shock Proteins ◽  
Host Response ◽  
Membrane Damage ◽  
Host Responses ◽  
Human Gut ◽  
Content Type ◽  
E Coli ◽  
Shock Proteins

ABSTRACT Measuring host-bacteriophage dynamics is an important approach to understanding bacterial survival functions and responses to infection. The model Microviridae bacteriophage φX174 is endemic to the human gut and has been studied for over 70 years, but the host response to infection has never been investigated in detail. To address this gap in our understanding of this important interaction within our microbiome, we have measured host Escherichia coli C proteomic and transcriptomic response to φX174 infection. We used mass spectrometry and RNA sequencing (RNA-seq) to identify and quantify all 11 φX174 proteins and over 1,700 E. coli proteins, enabling us to comprehensively map host pathways involved in φX174 infection. Most notably, we see significant host responses centered on membrane damage and remodeling, cellular chaperone and translocon activity, and lipoprotein processing, which we speculate is due to the peptidoglycan-disruptive effects of the φX174 lysis protein E on MraY activity. We also observe the massive upregulation of small heat shock proteins IbpA/B, along with other heat shock pathway chaperones, and speculate on how the specific characteristics of holdase protein activity may be beneficial for viral infections. Together, this study enables us to begin to understand the proteomic and transcriptomic host responses of E. coli to Microviridae infections and contributes insights to the activities of this important model host-phage interaction. IMPORTANCE A major part of the healthy human gut microbiome is the Microviridae bacteriophage, exemplified by the model φX174 phage, and their E. coli hosts. Although much has been learned from studying φX174 over the last half-century, until this work, the E. coli host response to infection has never been investigated in detail. We reveal the proteomic and transcriptomic pathways differentially regulated during the φX174 infection cycle and uncover the details of a coordinated cellular response to membrane damage that results in increased lipoprotein processing and membrane trafficking, likely due to the phage antibiotic-like lysis protein. We also reveal that small heat shock proteins IbpA/B are massively upregulated during infection and that these holdase chaperones are highly conserved across the domains of life, indicating that reliance on them is likely widespread across viruses.

Thermoprotective properties of small heat shock proteins from rice, tomato and Synechocystis sp. PCC6803 overexpressed in, and isolated from, Escherichia coli

2001 ◽  
Vol 28 (12) ◽  
pp. 1219
Author(s):  
Carl S. Pike ◽  
Joanne Grieve ◽  
Murray R. Badger ◽  
G. Dean Price
Keyword(s):  
Escherichia Coli ◽  
Heat Shock ◽  
Photosystem Ii ◽  
Heat Shock Proteins ◽  
Thermal Aggregation ◽  
E Coli ◽  
Synechococcus Sp ◽  
Shock Proteins ◽  
Synechocystis Sp

The present study forms part of a program investigating the role of small heat shock proteins (sHSPs) in the acquired and transgenic thermotolerance of the cyanobacterium Synechococcus PCC7942. The genes for three minimally related sHSPs, OsHSP from Oryza sativacytoplasm, tom111 from Lycopersicon esculentumchloroplasts, and 6803 HSP from Synechocystis sp. PCC6803, were cloned into the Escherichia coli vector pTrcHisA, so as to produce an N-terminal polyhistidine tag. The genes were transformed into E. coli and overexpressed. The tagged HSPs were purified (not completely in the case of tom111) by immobilised metal affinity chromatography. The native proteins exhibited a high degree of oligomerisation when analysed by size-exclusion chromatography. All three proteins were able to protect malate dehydrogenase (MDH) from in vitro thermal aggregation. They could also protect several soluble proteins in E. coli extracts from thermal aggregation in vitro, as well as protecting phycocyanin in extracts from Synechococcus sp. PCC7942. None of the proteins were able to protect photosystem II (measured as ΦPSII, the effective quantum fluorescence yield of PSII) of thylakoids isolated from Synechococcus sp. PCC7942 from heat damage in vitro, although in vivo, after acclimation, photosystem II did exhibit acquired thermotolerance.

scholarly journals Roles of the Escherichia coli Small Heat Shock Proteins IbpA and IbpB in Thermal Stress Management: Comparison with ClpA, ClpB, and HtpG In Vivo

1998 ◽  
Vol 180 (19) ◽  
pp. 5165-5172 ◽  
Author(s):  
Jeffrey G. Thomas ◽  
François Baneyx
Keyword(s):  
Escherichia Coli ◽  
Heat Shock ◽  
Heat Shock Proteins ◽  
High Temperatures ◽  
Small Heat Shock Proteins ◽  
Host Protein ◽  
E Coli ◽  
Growth Defects ◽  
Shock Proteins

ABSTRACT We have constructed an Escherichia coli strain lacking the small heat shock proteins IbpA and IbpB and compared its growth and viability at high temperatures to those of isogenic cells containing null mutations in the clpA, clpB, orhtpG gene. All mutants exhibited growth defects at 46°C, but not at lower temperatures. However, the clpA,htpG, and ibp null mutations did not reduce cell viability at 50°C. When cultures were allowed to recover from transient exposure to 50°C, all mutations except Δibpled to suboptimal growth as the recovery temperature was raised. Deletion of the heat shock genes clpB and htpGresulted in growth defects at 42°C when combined with thednaK756 or groES30 alleles, while the Δibp mutation had a detrimental effect only on the growth of dnaK756 mutants. Neither the overexpression of these heat shock proteins nor that of ClpA could restore the growth ofdnaK756 or groES30 cells at high temperatures. Whereas increased levels of host protein aggregation were observed indnaK756 and groES30 mutants at 46°C compared to wild-type cells, none of the null mutations had a similar effect. These results show that the highly conserved E. coli small heat shock proteins are dispensable and that their deletion results in only modest effects on growth and viability at high temperatures. Our data also suggest that ClpB, HtpG, and IbpA and -B cooperate with the major E. coli chaperone systems in vivo.

Increases of heat shock proteins and their mRNAs at high hydrostatic pressure in a deep-sea piezophilic bacterium, Shewanella violacea

Extremophiles ◽  
2015 ◽  
Vol 19 (4) ◽  
pp. 751-762 ◽  
Author(s):  
Hiroshi Sato ◽  
Kaoru Nakasone ◽  
Takao Yoshida ◽  
Chiaki Kato ◽  
Tadashi Maruyama
Keyword(s):  
Hydrostatic Pressure ◽  
Heat Shock ◽  
Heat Shock Proteins ◽  
Deep Sea ◽  
High Hydrostatic Pressure ◽  
Shewanella Violacea ◽  
Shock Proteins

Study of the inactivation of Escherichia coli and pectin methylesterase in mango nectar under selected high hydrostatic pressure treatments

2011 ◽  
Vol 17 (6) ◽  
pp. 541-547 ◽  
Author(s):  
D. Bermúdez-Aguirre ◽  
J. Ángel Guerrero-Beltrán ◽  
G.V. Barbosa-Cánovas ◽  
J. Welti-Chanes
Keyword(s):  
Escherichia Coli ◽  
High Pressure ◽  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Pectin Methylesterase ◽  
E Coli ◽  
Viable Cells ◽  
Plate Counts ◽  
The Media ◽  
Inactivation Effect

High hydrostatic pressure (HHP) was applied to fresh mango nectar (FMN) and sterilized mango nectar (SMN) to inactivate Escherichia coli and pectin methylesterase (PME). Pressure was applied at 275, 345 and 414 MPa. The come-up time (CUT) as well as 1, 2 and 4 min of treatment times were applied at the selected pressure to evaluate the inactivation effect on E. coli and PME. Total plate counts (TPC) were also evaluated in FMN. Results showed that mesophiles are inactivated in FMN to an important degree (up to 4 log) only with the CUT; the highest inactivation for mesophiles (7 log) was reported at 414 MPa after 4 min. Meanwhile, for E. coli 345 and 414 MPa after 2 and 1 min, respectively, were able to inactivate all viable cells in FMN. However, in SMN after 4 min at 275 MPa all cells of E. coli were also inactivated, showing the protective effect of the media between FMN and SMN. The PME showed its resistance to be inactivated with high pressure, showing the highest decrease in enzymatic activity (45%) after 4 min at 345 MPa but with an important activation at the highest pressure (414 MPa).

scholarly journals Carvacrol Induces Heat Shock Protein 60 and Inhibits Synthesis of Flagellin in Escherichia coli O157:H7

2007 ◽  
Vol 73 (14) ◽  
pp. 4484-4490 ◽  
Author(s):  
Sara A. Burt ◽  
Ruurd van der Zee ◽  
Ad P. Koets ◽  
Anko M. de Graaff ◽  
Frans van Knapen ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Heat Shock ◽  
Heat Shock Protein ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Sublethal Concentration ◽  
Exponential Growth Phase ◽  
Heat Shock Protein 60 ◽  
E Coli ◽  
Overnight Incubation

ABSTRACT The essential oils of oregano and thyme are active against a number of food-borne pathogens, such as Escherichia coli O157:H7. Carvacrol is one of the major antibacterial components of these oils, and p-cymene is thought to be its precursor in the plant. The effects of carvacrol and p-cymene on protein synthesis in E. coli O157:H7 ATCC 43895 cells were investigated. Bacteria were grown overnight in Mueller-Hinton broth with a sublethal concentration of carvacrol or p-cymene, and their protein compositions were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and confirmed by Western blotting. The presence of 1 mM carvacrol during overnight incubation caused E. coli O157:H7 to produce significant amounts of heat shock protein 60 (HSP60) (GroEL) (P < 0.05) and inhibited the synthesis of flagellin highly significantly (P < 0.001), causing cells to be aflagellate and therefore nonmotile. The amounts of HSP70 (DnaK) were not significantly affected. p-Cymene at 1 mM or 10 mM did not induce HSP60 or HSP70 in significant amounts and did not have a significant effect on flagellar synthesis. Neither carvacrol (0.3, 0.5, 0.8, or 1 mM) nor p-cymene (0.3, 0.5, or 0.8 mM) treatment of cells in the mid-exponential growth phase induced significant amounts of HSP60 or HSP70 within 3 h, although numerical increases of HSP60 were observed. Motility decreased with increasing concentrations of both compounds, but existing flagella were not shed. This study is the first to demonstrate that essential oil components induce HSP60 in bacteria and that overnight incubation with carvacrol prevents the development of flagella in E. coli O157:H7.

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