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Identification of an ECF sigma factor that facilitates electroactive biofilm formation by Shewanella oneidensis MR-1: Experiment 1

ABSTRACT: Shewanella oneidensis MR-1 was grown on electrodes in electrochemical flow cells (EFC), and the transcriptome profiles of electrode-attached MR-1 cells were compared under electrolyte-flow and static (non-flow) conditions. The results revealed that the SO_3096 gene encoding a putative extracytoplasmic function (ECF) sigma factor, as well as genes related to cyclic-di-guanosine monophosphate and flagella synthesis (e.g., SO_3556 and flrC) and c-type cytochrome maturation (dsbD), was significantly up-regulated under the electrolyte-flow condition. Compared to wild-type MR-1 (WT), a deletion mutant of SO_3096 (∆SO_3096) showed an impaired biofilm formation and a decreased current generation in EFC, suggesting that the ECF sigma factor encoded by this gene is involved in the regulation of biofilm formation and current generation under electrolyte-flow conditions. We also compared the transcriptome profiles of WT and ∆SO_3096 grown in EFC, confirming that many genes up-regulated under the electrolyte-flow condition, including dsbD, were down-regulated in ∆SO_3096. Transcription analysis using lacZ as a reporter gene showed that the promoter of dsbD is activated in the presence of SO_3096. Measurement of current generation by a dsbD-deletion mutant revealed that this gene is essential for electron transfer to electrodes. These results suggest that the SO_3096 protein serves as an ECF sigma factor that regulates cellular functions related to electroactive biofilm formation under electrolyte-flow conditions.

ORGANISM(S): Shewanella oneidensis MR-1 Shewanella oneidensis

PROVIDER: GSE142641 | GEO | 2020/06/01

REPOSITORIES: GEO

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Identification of an ECF sigma factor that facilitates electroactive biofilm formation by Shewanella oneidensis MR-1: Experiment 2

Project description:Shewanella oneidensis MR-1 was grown on electrodes in electrochemical flow cells (EFC), and the transcriptome profiles of electrode-attached MR-1 cells were compared under electrolyte-flow and static (non-flow) conditions. The results revealed that the SO_3096 gene encoding a putative extracytoplasmic function (ECF) sigma factor, as well as genes related to cyclic-di-guanosine monophosphate and flagella synthesis (e.g., SO_3556 and flrC) and c-type cytochrome maturation (dsbD), was significantly up-regulated under the electrolyte-flow condition. Compared to wild-type MR-1 (WT), a deletion mutant of SO_3096 (∆SO_3096) showed an impaired biofilm formation and a decreased current generation in EFC, suggesting that the ECF sigma factor encoded by this gene is involved in the regulation of biofilm formation and current generation under electrolyte-flow conditions. We also compared the transcriptome profiles of WT and ∆SO_3096 grown in EFC, confirming that many genes up-regulated under the electrolyte-flow condition, including dsbD, were down-regulated in ∆SO_3096. Transcription analysis using lacZ as a reporter gene showed that the promoter of dsbD is activated in the presence of SO_3096. Measurement of current generation by a dsbD-deletion mutant revealed that this gene is essential for electron transfer to electrodes. These results suggest that the SO_3096 protein serves as an ECF sigma factor that regulates cellular functions related to electroactive biofilm formation under electrolyte-flow conditions.

2020-06-01 | GSE142642 | GEO

Use of SWATH mass spectrometry for quantitative proteomic investigation of Shewanella oneidensis MR-1 biofilms grown on graphite cloth electrodes

Project description:. In this study we show successful use of SWATH-MS for quantitative proteomic analysis of a microbial electrochemically active biofilm. Shewanella oneidensis MR-1 was grown on carbon cloth electrodes under continuous anodic electrochemical polarizations in a bioelectrochemical system. Using lactate as the electron donor, anodes serving as terminal microbial electron acceptors were operated at three different electrode potentials (+0.71V, +0.21V & -0.19V vs. SHE) and the development of catalytic activity was monitored by measuring the current traces over time. Once maximum current was reached (usually within 21-29 hours) the electrochemical systems were shut off and biofilm proteins were extracted from the electrodes for proteomic assessment.

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Regulation of Energy Metabolism by the Extracytoplasmic Function (ECF) σ factors of Arcobacter butzleri

Project description:The extracytoplasmic function (ECF) σ factors are fundamental for bacterial adaptation to distinct environments and for survival under different stress conditions. The emerging pathogen Arcobacter butzleri possesses seven putative pairs of σ/anti-σ factors belonging to the ECF family. Here, we report the identification of the genes regulated by five out of the seven A. butzleri ECF σ factors. Three of the ECF σ factors play an apparent role in transport, energy generation and the maintainance of redox balance. Several genes like the nap, sox and tct genes are regulated by more than one ECF σ factor indicating that the A. butzleri ECF σ factors form a network of overlapping regulons. In contrast to other eubacteria, these A. butzleri ECF regulons appear to primarily regulate responses to changing environments in order to meet metabolic needs instead of an obvious role in stress adaptation.

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Bacillus subtilis 168, WT vs. ΔMWX, WT vs. Δ7ECF, Δ7ECF vs. ΔMWX.

Project description:Bacillus subtilis encodes seven extracytoplasmic function (ECF) sigma factors. Three (sigma M, sigma W and simga X) mediate responses to cell envelope active antibiotics. The functions of sigma Y, sigma Z, sigma V, and YlaC remain largely unknown, and strong inducers of these sigma factors and their regulons have yet to be defined. Here, we define transcriptomic and phenotypic differences under non-stress conditions between strains carrying deletions in all seven ECF sigma factor genes (Δ7ECF), a sigMWX triple mutant (∆MWX), and the parental 168 strain. Our results identify >80 genes as at least partially dependent on ECF sigma factors and, as expected, most of these are dependent on sigma M, sigma W or sigma X which are active at a significant basal level during growth. Several genes, including the eps operon encoding enzymes for exopolysaccharide (EPS) production, were decreased in expression in Δ7ECF but affected little if at all in ΔMWX. Consistent with this observation, Δ7ECF (but not ∆MWX) showed reduced biofilm formation. Extending previous observations, we also note that ∆MWX is sensitive to a variety of antibiotics and Δ7ECF is either as sensitive as, or slightly more sensitive than, the ΔMWX strain to these stressors. These findings emphasize the overlapping nature of the seven ECF s factor regulons in B. subtilis, confirm that three of these (sigma M, W or X) play the dominant role in conferring intrinsic resistance to antibiotics, and provide initial insights into the roles of the remaining ECF sigma factors.

2010-07-21 | GSE22930 | GEO

The temperature-regulation of Pseudomonas aeruginosa cmaX-cfrX-cmpX operon reveals an intriguing molecular network involving the sigma factors AlgU and SigX

Project description:Pseudomonas aeruginosa is a highly adaptable Gram-negative opportunistic pathogen, notablydue to its large number of transcription regulators. The extracytoplasmic sigma factor (ECF sigma AlgU, responsible for alginate biosynthesis, is also involved in responses to cell wall stress and heat shock via the RpoH alternative sigma factor. The SigX ECF sigma emerged as a major regulator involved in the envelope stress response via membrane remodelling, 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 ECF sigma 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.

2020-10-05 | PXD021299 | Pride

Bacillus subtilis 168, WT vs. ΔMWX, WT vs. Δ7ECF, Δ7ECF vs. ΔMWX.

2010-07-21 | E-GEOD-22930 | biostudies-arrayexpress

Expression profiling analyses of ECF sigma factor PG1660 mutant of Porphyromonas gingivalis W83

Project description:To study the expression profile of ECF sigma factor PG1660 mutant under anaerobic conditions and hydrogen peroxide stress conditions compared to the wild-type W83 by using DNA-microarray. The role of ECF sigma factor PG1660 involved in oxidative stress was published Yuetan Dou, Devon Osbourne, Rachelle McKenzie, Hansel M Fletcher. (2010) Involvement of extracytoplasmic function sigma factors in virulence regulation in Porphyromonas gingivalis W83. FEMS Microbiology Letter, 312(1):24-32.

2017-09-14 | GSE103838 | GEO

Identification of an ECF sigma factor that facilitates electroactive biofilm formation by Shewanella oneidensis MR-1

Project description:Identification of an ECF sigma factor that facilitates electroactive biofilm formation by Shewanella oneidensis MR-1

| PRJNA597846 | ENA

SigX ECF sigma factor deletion mutant expression profile in Pseudomonas aeruginosa in M9 minimal medium (M9G)

Project description:Analysis of a SigX knockout mutant of Pseudomonas aeruginosa H103 strain in minimal medium with glucose as carbon source (M9G). SigX, one of the 19 extra-cytoplasmic function sigma factors of P. aeruginosa, was only known to be involved in transcription of the gene encoding the major outer membrane protein OprF in Pseudomonas aeruginosa. Deletion of the ECF sigma factor sigX gene provide insights into the SigX role in several virulence and biofilm- related phenotypes in Pseudomonas aeruginosa.

2014-01-01 | GSE51076 | GEO

Electrochemical selection and characterization of a high current-generating Shewanella oneidensis mutant with altered cell-surface morphology and biofilm-related gene expression

Project description:Shewanella oneidensis MR-1 exhibits extracellular electron transfer (EET) activity that is influenced by various cellular components, including outer-membrane cytochromes, cell-surface polysaccharides (CPS), and regulatory proteins. Here, a random transposon-insertion mutant library of S. oneidensis MR-1 was screened after extended cultivation in electrochemical cells (ECs) with a working electrode poised at +0.2 V (vs. Ag/AgCl) to isolate mutants that adapted to electrode-respiring conditions and identify as-yet-unknown EET-related factors. Several mutants isolated from the enrichment culture exhibited rough morphology and extraordinarily large colonies on agar plates compared to wild-type MR-1. One of the isolated mutants, designated strain EC-2, produced 90% higher electric current than wild-type MR-1 in ECs and was found to have a transposon inserted in the SO_1860 (uvrY) gene, which encodes a DNA-binding response regulator of the BarA/UvrY two-component regulatory system. However, an in-frame deletion mutant of SO_1860 (∆SO_1860) did not exhibit a similar level of current generation as that of EC-2, suggesting that the enhanced current-generating capability of EC-2 was not simply due to the disruption of SO_1860. In both EC-2 and ∆SO_1860, the transcription of genes related to CPS synthesis was decreased compared to wild-type MR-1, suggesting that CPS negatively affects current generation. In addition, transcriptome analyses revealed that a number of genes, including those involved in biofilm formation, were differentially expressed in EC-2 compared to those in ∆SO_1860. The present results indicate that the altered expression of the genes related to CPS biosynthesis and biofilm formation is associated with the distinct morphotype and high current-generating capability of strain EC-2, suggesting an important role of these genes in determining the EET activity of S. oneidensis.

2014-07-14 | GSE50443 | GEO

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