Галерея 579495
Галерея 579495
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1 Laboratoire de Microbiologie Signaux et Microenvironnement (LMSM) EA 4312, Normandie Université, Université de Rouen Normandie, Centre de Sécurité Sanitaire de Normandie, Evreux, France.
2 Institute of Functional Interfaces, Karlsruhe Institute of Technology, Karlsruhe, Germany.
3 Laboratoire de Biotechnologie et Chimie Marines (LBCM) EA3884, IUEM, Université de Bretagne-Sud, Lorient, France.
Emeline Bouffartigues et al.
Front Microbiol .
2020 .
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Abstract
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Affiliations
1 Laboratoire de Microbiologie Signaux et Microenvironnement (LMSM) EA 4312, Normandie Université, Université de Rouen Normandie, Centre de Sécurité Sanitaire de Normandie, Evreux, France.
2 Institute of Functional Interfaces, Karlsruhe Institute of Technology, Karlsruhe, Germany.
3 Laboratoire de Biotechnologie et Chimie Marines (LBCM) EA3884, IUEM, Université de Bretagne-Sud, Lorient, France.
Effect of HS and CS on cmaX-cfrX-cmpX expression P. aeruginosa H103. Histograms show the means of normalized expression for each gene in HS (A) or CS (B) relatively to the control condition at 37°C as calibrator. Error bars indicate SEM of mean, n = 4. Significant differences between the sets of normalized expression obtained in each stress condition were calculated against those obtained at 37°C using a two-tailed t -test, and P- value are indicated by * or *** if < 0.05 or 0.0001, respectively. The symbol ° is indicated for non-significant results.
At least two promoter regions control the transcription of cmaX, cfrX , and cmpX in P. aeruginosa H103. (A) Schematic representation of the genomic environment of the predicted operon cmaX-cfrX-cmpX of P. aeruginosa . Large arrows indicate the direction of transcription of the ORFs, loops show the position of the putative rho-independent transcription terminators (T1 and T2), and the vertical black rectangles represent the two BPROM predicted promoters. (B) Sequences of the two putative promoters P cmaX and P cfrX . Predicted –35 and –10 motifs using BPROM are in bold case and underlined, the DNA-binding motif for AlgU is surrounded in black and TSSs are indicated by arrows. (C) Electrophoregrams obtained from 5′RACE-PCR experiments confirming the promoters upstream cmaX and cfrX genes. (D) Schematic representation of the construct harboring the pAB cmaX or pAB cfrX vectors. (A–C) * and ** correspond to the two distinct promoter regions. (E) Relative activity of the promoter regions upstream cmaX or cfrX in P. aeruginosa H103 during growth in LB medium at 37°C. Reporter constructs contain the P cmaX (construct 1, pAB cmaX ) or the P cfrX (construct 2, pAB cfrX ) promoter fused to the lux CDABE operon. The large white rectangle indicates the lux CDABE cassette, the vertical black rectangles represent promoters and a line shows the DNA sequence directly upstream cmaX or cfrX ORF. The position of the last nucleotide (relatively to the translational initiation site) included within the cloned DNA fragment is highlighted. The mean profile for growth and reporter assay from H103 transformed with construct 1 or 2 are shown and correspond to 3 independent experiments, ± SD. Transformants growth is presented in gray and the transcriptional activity pattern in black.
Involvement of AlgU in cmaX, cfrX, and cmpX regulation under HS. (A) Activity pattern of the promoter region upstream cmaX in an algU mutant (black squares) or in H103 (dashed line) during growth in LB at 37°C of the H103 wild-type strain (black dashed line) or Δ algU mutant (gray squares). n = 3, error bars indicate SD. (B) Histograms show the means of normalized expression relatively to H103 at 37°C as calibrator (white bars) for some known algU targets (red frame) and the studied operon (green frame) in H103 (red line) and in Δ algU mutant exposed (dashed back bars) or not (dashed white bars) to HS. NT, not treated. Errors bars indicate SEM, n = 4. Significant differences between the sets of normalized expression obtained in HS condition were calculated against those obtained at 37°C for H103 using a two-tailed t -test and P- value, which are indicated by *, **, if < 0.05, 0.001, respectively. The symbol ° is indicated for non-significant results.
sigX is up-regulated in response to CS and valinomycin promoting membrane stiffening. (A)…
sigX is up-regulated in response to CS and valinomycin promoting membrane stiffening. (A) Effect of thermal shocks on sigX expression and activity. Gray and black bars represent the mean normalized expression for sigX and its targets after CS or HS relatively to those obtained for H103 at 37°C (dotted and white bars). NT, not treated. Errors bars indicate SEM, n = 4. Significant differences between the sets of normalized expression obtained in each stress condition were calculated against those obtained at 37°C using a two-tailed t -test and P -value, which are indicated by *, **, or *** if < 0.05, 0.001, or 0.0001, respectively. The symbol ° is indicated for non-significant results. (B) TMA-DPH and DPH anisotropies was used to evaluate the effect of 45 μM valinomycin on membrane fluidity 2 h after the addition of the membrane acting-agent to mid-log phase cultures. Values obtained from unexposed cultures are indicated NT: not treated. Box plots correspond to each sets of data from three independent experiments. Means, medians and min/max anisotropies values are indicated by squares, lines and crosses, respectively. Significant differences between each condition were calculated using a two-tailed t -test and P -value, which are indicated by * if < 0.05. The symbol ° is indicated for non-significant results. (C) Relative expression of sigX after 2 h of exposure to 45 μM valinomycin. NT, not treated. Error bars indicate SEM, n = 3, P -value is calculated by two-tailed t -test and is indicated by ** if < 0.001.
Involvement of SigX in cmaX , cfrX , and cmpX and amrZ regulation under CS. (A) Growth of H103 (black squares, dashed line) and PAOSX sigX mutant (gray squares) in LB at 37°C and activity pattern of the promoter region upstream cfrX in H103 (dashed line) and PAOSX (black circles) background. N = 3, error bars indicate SD. (B) Means of normalized expression relatively to H103 at 37°C (dotted white bars) of sigX and its targets (blue frame) and genes belonging to the operon (green frame), in H103 (red line) and in PAOSX exposed (dashed back bars) or not (dashed white bars) to CS. NT, not treated. (C) White bars show mean normalized amrZ expression (relative to H103 at 37°C) in H103 or PAOSX exposed or not to CS. (B,C) Error bars indicate SEM for n = 4, P -value were calculated by two-tailed t -test and are indicated by *, **, or *** if < 0.05, 0.001, or 0.0001, respectively. The symbol ° is indicated for non-significant results.
Predicted model depicting the molecular mechanisms involved in the regulation of cmaX-cfrX-cmpX in response to HS and CS. Red and green lines correspond to negative or positive regulation, respectively.
Bouffartigues E, Gicquel G, Bazire A, Bains M, Maillot O, Vieillard J, Feuilloley MG, Orange N, Hancock RE, Dufour A, Chevalier S.
Bouffartigues E, et al.
J Bacteriol. 2012 Aug;194(16):4301-11. doi: 10.1128/JB.00509-12. Epub 2012 Jun 8.
J Bacteriol. 2012.
PMID: 22685281
Free PMC article.
Chevalier S, Bouffartigues E, Bazire A, Tahrioui A, Duchesne R, Tortuel D, Maillot O, Clamens T, Orange N, Feuilloley MGJ, Lesouhaitier O, Dufour A, Cornelis P.
Chevalier S, et al.
Biochim Biophys Acta Gene Regul Mech. 2019 Jul;1862(7):706-721. doi: 10.1016/j.bbagrm.2018.04.008. Epub 2018 May 2.
Biochim Biophys Acta Gene Regul Mech. 2019.
PMID: 29729420
Review.
Brinkman FS, Schoofs G, Hancock RE, De Mot R.
Brinkman FS, et al.
J Bacteriol. 1999 Aug;181(16):4746-54. doi: 10.1128/JB.181.16.4746-4754.1999.
J Bacteriol. 1999.
PMID: 10438740
Free PMC article.
Blanka A, Schulz S, Eckweiler D, Franke R, Bielecka A, Nicolai T, Casilag F, Düvel J, Abraham WR, Kaever V, Häussler S.
Blanka A, et al.
J Bacteriol. 2014 Jan;196(2):345-56. doi: 10.1128/JB.01034-13. Epub 2013 Nov 1.
J Bacteriol. 2014.
PMID: 24187091
Free PMC article.
Tortuel D, Tahrioui A, Rodrigues S, Cambronel M, Boukerb AM, Maillot O, Verdon J, Bere E, Nusser M, Brenner-Weiss G, David A, Azuama OC, Feuilloley MGJ, Orange N, Lesouhaitier O, Cornelis P, Chevalier S, Bouffartigues E.
Tortuel D, et al.
Microorganisms. 2020 Oct 30;8(11):1700. doi: 10.3390/microorganisms8111700.
Microorganisms. 2020.
PMID: 33143386
Free PMC article.
Tribelli PM, López NI.
Tribelli PM, et al.
Appl Microbiol Biotechnol. 2022 Dec;106(23):7699-7709. doi: 10.1007/s00253-022-12243-z. Epub 2022 Oct 22.
Appl Microbiol Biotechnol. 2022.
PMID: 36271255
Review.
Chevalier S, Bouffartigues E, Tortuel D, David A, Tahrioui A, Labbé C, Barreau M, Tareau AS, Louis M, Lesouhaitier O, Cornelis P.
Chevalier S, et al.
Adv Exp Med Biol. 2022;1386:147-184. doi: 10.1007/978-3-031-08491-1_6.
Adv Exp Med Biol. 2022.
PMID: 36258072
Louis M, Tahrioui A, Verdon J, David A, Rodrigues S, Barreau M, Manac'h M, Thiroux A, Luton B, Dupont C, Calvé ML, Bazire A, Crépin A, Clabaut M, Portier E, Taupin L, Defontaine F, Clamens T, Bouffartigues E, Cornelis P, Feuilloley M, Caillon J, Dufour A, Berjeaud JM, Lesouhaitier O, Chevalier S.
Louis M, et al.
Microorganisms. 2022 Sep 5;10(9):1788. doi: 10.3390/microorganisms10091788.
Microorganisms. 2022.
PMID: 36144390
Free PMC article.
Marcos-Torres FJ, Moraleda-Muñoz A, Contreras-Moreno FJ, Muñoz-Dorado J, Pérez J.
Marcos-Torres FJ, et al.
Int J Mol Sci. 2022 Mar 25;23(7):3601. doi: 10.3390/ijms23073601.
Int J Mol Sci. 2022.
PMID: 35408957
Free PMC article.
Review.
Cornelis P, Tahrioui A, Lesouhaitier O, Bouffartigues E, Feuilloley M, Baysse C, Chevalier S.
Cornelis P, et al.
Biometals. 2022 Feb 16. doi: 10.1007/s10534-022-00369-6. Online ahead of print.
Biometals. 2022.
PMID: 35171432
Review.
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Pseudomonas aeruginosa is a highly adaptable Gram-negative opportunistic pathogen, notably due to its large number of transcription regulators. The extracytoplasmic sigma factor (ECFσ) AlgU, responsible for alginate biosynthesis, is also involved in responses to cell wall stress and heat shock via the RpoH alternative σ factor. The SigX ECFσ emerged as a major regulator involved in the envelope stress response via membrane remodeling, virulence and biofilm formation. However, their functional interactions to coordinate the envelope homeostasis in response to environmental variations remain to be determined. The regulation of the putative cmaX-cfrX-cmpX operon located directly upstream sigX was investigated by applying sudden temperature shifts from 37°C. We identified a SigX- and an AlgU- dependent promoter region upstream of cfrX and cmaX , respectively. We show that cmaX expression is increased upon heat shock through an AlgU-dependent but RpoH independent mechanism. In addition, the ECFσ SigX is activated in response to valinomycin, an agent altering the membrane structure, and up-regulates cfrX-cmpX transcription in response to cold shock. Altogether, these data provide new insights into the regulation exerted by SigX and networks that are involved in maintaining envelope homeostasis.
Keywords:
Pseudomonas aeruginosa; cell wall stress ECF sigma factor; membrane fluidity; regulation of transcription; temperature.
Copyright © 2020 Bouffartigues, Si Hadj Mohand, Maillot, Tortuel, Omnes, David, Tahrioui, Duchesne, Azuama, Nusser, Brenner-Weiss, Bazire, Connil, Orange, Feuilloley, Lesouhaitier, Dufour, Cornelis and Chevalier.
Effect of HS and CS on cmaX-cfrX-cmpX expression P. aeruginosa H103. Histograms show…
At least two promoter regions control the transcription of cmaX, cfrX , and…
Involvement of AlgU in cmaX, cfrX, and cmpX regulation under HS. (A) Activity…
Involvement of SigX in cmaX , cfrX , and cmpX and amrZ regulation…
Predicted model depicting the molecular…
Predicted model depicting the molecular mechanisms involved in the regulation of cmaX-cfrX-cmpX in…
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