Breaking through the stress barrier: the role of BolA in Gram-negative survival

Guinote, Inês Batista; Moreira, Ricardo Neves; Barahona, Susana; Freire, Patrick; Vicente, Miguel; Arraiano, Cecília M.

doi:10.1007/s11274-014-1702-4

Cited by 26 publications

(29 citation statements)

References 54 publications

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“…In Escherichia coli, overexpression of bolA leads to a rounded morphology and induction of biofilm formation (38,39). bolA is normally expressed in the stationary phase but can be induced under stress conditions; therefore, the increased expression of bolA and its regulator, rpoS, under the stress condition of antibiotic therapy is consistent with previous studies (38,39). Also potentially affecting cell morphology could be the downregulation of genes involved in the biosynthesis of peptidoglycan, including murDGP, mraYZ, uppP, and dacC.…”

Section: Discussionmentioning

confidence: 99%

Sublethal Concentrations of Carbapenems Alter Cell Morphology and Genomic Expression of Klebsiella pneumoniae Biofilms

Laar

Chen²,

You

et al. 2015

Antimicrob Agents Chemother

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b Klebsiella pneumoniae, a Gram-negative bacterium, is normally associated with pneumonia in patients with weakened immune systems. However, it is also a prevalent nosocomial infectious agent that can be found in infected surgical sites and combat wounds. Many of these clinical strains display multidrug resistance. We have worked with a clinical strain of K. pneumoniae that was initially isolated from a wound of an injured soldier. This strain demonstrated resistance to many commonly used antibiotics but sensitivity to carbapenems. This isolate was capable of forming biofilms in vitro, contributing to its increased antibiotic resistance and impaired clearance. We were interested in determining how sublethal concentrations of carbapenem treatment specifically affect K. pneumoniae biofilms both in morphology and in genomic expression. Scanning electron microscopy showed striking morphological differences between untreated and treated biofilms, including rounding, blebbing, and dimpling of treated cells. Comparative transcriptome analysis using RNA sequencing (RNA-Seq) technology identified a large number of open reading frames (ORFs) differentially regulated in response to carbapenem treatment at 2 and 24 h. ORFs upregulated with carbapenem treatment included genes involved in resistance, as well as those coding for antiporters and autoinducers. ORFs downregulated included those coding for metal transporters, membrane biosynthesis proteins, and motility proteins. Quantitative real-time PCR validated the general trend of some of these differentially regulated ORFs. Treatment of K. pneumoniae biofilms with sublethal concentrations of carbapenems induced a wide range of phenotypic and gene expression changes. This study reveals some of the mechanisms underlying how sublethal amounts of carbapenems could affect the overall fitness and pathogenic potential of K. pneumoniae biofilm cells.

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Section: Discussionmentioning

confidence: 99%

Sublethal Concentrations of Carbapenems Alter Cell Morphology and Genomic Expression of Klebsiella pneumoniae Biofilms

Laar

Chen²,

You

et al. 2015

Antimicrob Agents Chemother

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“…Starved bacterial cells display tendency towards grouping: the failure to separate after division, described in (Wainwright et al, 1999), results in filamentous growth which conforms to the reports of UspE-dependent cell aggregation (Nachin, Nannmark & Nyström, 2005), and transcriptional factor BolA-mediated biofilm formation (Guinote et al, 2014;Dressaire et al, 2015) (both of the proteins were upregulated by starvation). Our 2D annotation analysis unequivocally showed that starved cells are enriched in 'Biological adhesion' GO term and thus Formation of cellular aggregates, such as biofilms, could serve as a defense mechanism against antibiotics as described in (Nguyen et al, 2011;Bernier et al, 2013).…”

Section: Discussionsupporting

confidence: 82%

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Starvation is a complex adaptive response to insufficiency of nutrients that has been known to implicate a number of stress networks, and modulate pathogenicity and antibiotic resistance in bacteria. However, naturally occurring abrupt elimination of nutrients and prolonged periods of their complete absence, e.g. when bacteria are placed in natural or artificial water reservoirs, are qualitatively different from in-culture late stationary phase energy source diminution. Despite the obvious importance of proteomic investigation of bacteria exposed to nutrient deficiency, no comprehensive study on the subject has been published. In order to address the said shortage of knowledge, we decided to quantitatively look into the proteome-level alterations elicited by the complete lack of nutrients that constitute a viable source of carbon, i.e. carbon starvation, in the Escherichia coli HT115-derived SLE1 strain cells using the combination of label-free and SILAC-based proteomics. As a result, we obtained protein ratios for 1,757 and 1,241 protein groups for each technique respectively, 2D-annotated the quantifiable proteins present in both datasets, identified over-and underrepresented Gene Ontology terms, and isolated protein groups ≥2-fold up-and downregulated in response to carbon starvation (44 and 36 protein groups respectively). We observed upregulation of proteins implicated in various stress-related networks, most notably those that constitute the Gene Ontology term 'Biological adhesion', as well as various terms related to stress. Additionally, we identified several uncharacterized proteins, and our report is the first to ascribe them to a stress-induced proteome. Our data are available via ProteomeXchange with identifier PXD003255 and DOI:10.6019/PXD003255. PeerJ PrePrints | https://doi.org/10.7287/peerj.preprints.1572v1 | CC-BY 4.0 Open Access | rec1 2 3 4 5 6 7 8 9 PeerJ PrePrints | https://doi.org/10.7287/peerj.preprints.1572v1 | CC-BY 4.0 Open Access | rec ABSTRACTStarvation is a complex adaptive response to insufficiency of nutrients that has been known to implicate a number of stress networks, and modulate pathogenicity and antibiotic resistance in bacteria. However, naturally occurring abrupt elimination of nutrients and prolonged periods of their complete absence, e.g. when bacteria are placed in natural or artificial water reservoirs, are qualitatively different from in-culture late stationary phase energy source diminution. Despite the obvious importance of proteomic investigation of bacteria exposed to nutrient deficiency, no comprehensive study on the subject has been published. In order to address the said shortage of knowledge, we decided to quantitatively look into the proteome-level alterations elicited by the complete lack of nutrients that constitute a viable source of carbon, i.e. carbon starvation, in the Escherichia coli HT115-derived SLE1 strain cells using the combination of label-free and SILAC-based proteomics. As a result, we obtained protein ratios for 1,757 and 1,241 protein groups f...

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“…Since BolA family proteins have been shown to participate in stress response pathways (2), we explored whether the absence of ibaG altered V. cholerae survival following exposure to a range of environmental stresses. Given results from studies of E. coli IbaG, we first assessed whether V. cholerae IbaG modulates growth or survival under acidic conditions.…”

Section: Resultsmentioning

confidence: 99%

“… A . Coomassie Blue stained gel of proteins recovered after TAP purification from cell extracts of V. cholerae producing TAP-TAG only (1) or IbaG-TAP-TAG (2). Bands of interest were analyzed by mass spectrometry.…”

Section: Resultsmentioning

confidence: 99%

“…The BolA protein family is widely conserved across gram-negative bacteria and eukaryotes (1). These proteins have been linked to a range of cellular phenotypes, including cell morphology, membrane permeability, motility, and biofilm formation (2). BolA-like proteins have a class II KH fold related to that of the OsmC hyperperoxide reductase (3) that includes a helix-turn helix domain (HTH) (1).…”

Section: Introductionmentioning

confidence: 99%

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AVibrio choleraeBolA-like protein is required for proper cell shape and cell envelope integrity

Fleurie

Zoued

Álvarez

et al. 2019

Preprint

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25BolA family proteins are conserved in gram-negative bacteria and many eukaryotes. 26 While diverse cellular phenotypes have been linked to this protein family, the molecular 27 pathways through which these proteins mediate their effects are not well-described. Here, 28 we investigated the role of BolA family proteins in Vibrio cholerae, the cholera pathogen. 29 Like Escherichia coli, V. cholerae encodes two BolA proteins, BolA and IbaG. However, 30 in marked contrast to E. coli, where bolA is linked to cell shape and ibaG is not, in V. 31 cholerae, bolA mutants lack morphological defects, whereas ibaG proved critical for the 32 generation and/or maintenance of the pathogen's morphology. Notably, the bizarre-33 shaped, multi-polar, elongated and wide cells that predominated in exponential phase 34 ΔibaG V. cholerae cultures were not observed in stationary phase cultures. The V. 35 cholerae ΔibaG mutant exhibited increased sensitivity to cell envelope stressors, 36 including cell wall acting antibiotics and bile, and was defective in intestinal colonization. 37 ΔibaG V. cholerae had reduced peptidoglycan and lipid II and altered outer membrane 38 lipids, likely contributing to the mutant's morphological defects and sensitivity to envelope 39 stressors. Transposon-insertion sequencing analysis of ibaG's genetic interactions 40 suggested that ibaG is involved in several processes involved in the generation and 41 homeostasis of the cell envelope. Furthermore, co-purification studies revealed that IbaG 42 interacts with proteins containing iron-sulfur clusters or involved in their assembly. 43 Collectively, our findings suggest that V. cholerae IbaG controls cell morphology and cell 44 envelope integrity through its role in biogenesis or trafficking of iron-sulfur cluster proteins. 45 46 47 3 Importance 48BolA-like proteins are conserved across prokaryotes and eukaryotes. These proteins 49 have been linked to a variety of phenotypes, but the pathways and mechanisms through 50 which they act have not been extensively characterized. Here, we unraveled the role of 51 the BolA-like protein IbaG in the cholera pathogen Vibrio cholerae. The absence of IbaG 52 was associated with dramatic changes in cell morphology, sensitivity to envelope 53 stressors, and intestinal colonization defects. IbaG was found to be required for 54 biogenesis of several components of the V. cholerae cell envelope and to interact with 55 numerous iron-sulfur cluster containing proteins and factors involved in their assembly. 56 Thus, our findings suggest that IbaG governs V. cholerae cell shape and cell envelope 57 homeostasis through its effects on iron-sulfur proteins and associated pathways. The 58 diversity of processes involving iron-sulfur containing proteins is likely a factor underlying 59 the range of phenotypes associated with BolA family proteins. 60 61 4 62The BolA protein family is widely conserved across gram-negative bacteria and 63 eukaryotes (1). These proteins have been linked to a range ...

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Breaking through the stress barrier: the role of BolA in Gram-negative survival

Cited by 26 publications

References 54 publications

Sublethal Concentrations of Carbapenems Alter Cell Morphology and Genomic Expression of Klebsiella pneumoniae Biofilms

Sublethal Concentrations of Carbapenems Alter Cell Morphology and Genomic Expression of Klebsiella pneumoniae Biofilms

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AVibrio choleraeBolA-like protein is required for proper cell shape and cell envelope integrity

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