Proteomic analysis of Escherichia coli detergent-resistant membranes (DRM)

Guzmán-Flores, José E.; Steinemann-Hernández, Lidia; Vara, Luis E. González de la; Gavilanes‐Ruíz, Marina; Romeo, Tony; Álvarez, Adrián F.; Georgellis, Dimitris

doi:10.1371/journal.pone.0223794

Cited by 11 publications

(25 citation statements)

References 51 publications

(77 reference statements)

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“…It is conceivable that under stress conditions, for example the exposure to high temperature when the lapC mRNA is made more, the LapB/FtsH activity is repressed when LapC is more. Supporting our results of increased accumulation of LapC at high temperature are independent proteomic analysis data of total E. coli proteome from high temperature or other stress conditions [63,64]. In line with such arguments are findings that LpxC is more rapidly degraded at lower temperatures than at higher temperatures correlating it with doubling time of E. coli [56].…”

Section: Discussionsupporting

confidence: 86%

Regulation of the First Committed Step in Lipopolysaccharide Biosynthesis Catalyzed by LpxC Requires the Essential Protein LapC (YejM) and HslVU Protease

Biernacka

Gorzelak

Klein

et al. 2020

IJMS

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We previously showed that lipopolysaccharide (LPS) assembly requires the essential LapB protein to regulate FtsH-mediated proteolysis of LpxC protein that catalyzes the first committed step in the LPS synthesis. To further understand the essential function of LapB and its role in LpxC turnover, multicopy suppressors of ΔlapB revealed that overproduction of HslV protease subunit prevents its lethality by proteolytic degradation of LpxC, providing the first alternative pathway of LpxC degradation. Isolation and characterization of an extragenic suppressor mutation that prevents lethality of ΔlapB by restoration of normal LPS synthesis identified a frame-shift mutation after 377 aa in the essential gene designated lapC, suggesting LapB and LapC act antagonistically. The same lapC gene was identified during selection for mutations that induce transcription from LPS defects-responsive rpoEP3 promoter, confer sensitivity to LpxC inhibitor CHIR090 and a temperature-sensitive phenotype. Suppressors of lapC mutants that restored growth at elevated temperatures mapped to lapA/lapB, lpxC and ftsH genes. Such suppressor mutations restored normal levels of LPS and prevented proteolysis of LpxC in lapC mutants. Interestingly, a lapC deletion could be constructed in strains either overproducing LpxC or in the absence of LapB, revealing that FtsH, LapB and LapC together regulate LPS synthesis by controlling LpxC amounts.

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Section: Discussionsupporting

confidence: 86%

Regulation of the First Committed Step in Lipopolysaccharide Biosynthesis Catalyzed by LpxC Requires the Essential Protein LapC (YejM) and HslVU Protease

Biernacka

Gorzelak

Klein

et al. 2020

IJMS

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“…We first tested AcrA, the efflux pump involved in the transport of a wide range of substrates including aminoglycosides (45) and YidC, an essential inner membrane protein required for proper insertion of integral inner membrane proteins (46). Both proteins were indeed previously identified in E. coli inner-membrane DRM fractions often biochemically associated with FMM and shown to display HflC-type pattern of polar localization when fused to fluorescent proteins and expressed from plasmids (35,47). However, when expressed from their native chromosomal context, we could not detect any distinct AcrA-GFP nor YidC-GFP localization nor any significant co-localisation with HflC in exponential or stationary phase conditions (Sup.…”

Section: Yajc Interacts and Colocalizes With Hflc And Contributes To ...mentioning

confidence: 99%

“…HflC and QmcA are detected in E. coli membrane fractions resistant to solubilization by non-ionic detergents (detergent-resistant membranes or DRM) that are often used as - debatable - proxies for FMMs (31–34). Moreover, fluorescent microscopy showed that E. coli SPFH proteins HflC and QmcA are localized in discrete polar or lateral membrane foci (35). This raised the possibility that E. coli SPFH proteins could localize in inner-membrane FMMs and regulate important cellular processes (36).…”

Section: Introductionmentioning

confidence: 99%

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Escherichia coliSPFH membrane microdomain proteins HflKC contribute to aminoglycoside and oxidative stress tolerance

Wessel

Yoshii

Reder³

et al. 2022

Preprint

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Eukaryotic cells segregate many membrane-dependent functions into membrane microdomains also known as lipid rafts. These domains are enriched in polyisoprenoid lipids and in scaffolding proteins belonging to the Stomatin, Prohibitin, Flotillin, and HflK/C (SPFH) protein superfamily, which are also found in prokaryotes. Whereas Gram-positive bacteria were also shown to possess functional membrane microdomains (FMMs) structurally and functionally similar to eukaryotic lipid rafts, little is still known about Gram-negative bacteria FMMs. Escherichia coli K12 possesses 4 SPFH proteins, YqiK, QmcA, HflK, and HflC, previously shown to localize in discrete polar or lateral inner-membrane locations, raising the possibility that E. coli SPFH proteins could contribute to the assembly of inner-membrane FMMs regulating cellular processes. Here we studied the determinants of the native, chromosomal QmcA and HflC cell localization using a domain swap analysis and fluorescent and super-resolution microscopy. We showed that full QmcA and HflC protein is required for achieving their native inner-membrane localization and that impairing the synthesis of cardiolipin and isoprenoid lipids known to associate with FMMs alters QmcA and HflC localization pattern. Finally, using Biolog phenotypic arrays, we showed that a mutant lacking all SPFH genes displayed increased sensitivity to aminoglycosides and oxidative stress. This phenotype is exclusively due to the absence of HflKC and a cross-linking and mass spectrometry analysis showed that YajC, a SecDF translocon accessory protein, interacts with HflC and also contributes to E. coli stress tolerance. Our study therefore provides insights into the function and interactions associated with SPFH proteins in E. coli FMMs.

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“…2b, Table S2). Apart from phage-related genes, those in the chromosome were involved in metabolism (acetolactate synthase, ABC transporter substrate-binding protein), DNA replication (DNA primase and ATP-dependent helicase), signalling (hybrid sensor histidine kinase (rpfC), two-component system response regulator (rpfG) [58] and SPFH/Band 7/PHB domain protein [59,60]), gene expression (helix-turn-helix domain containing protein, transcriptional regulator Crp), cell wall growth/lysis [61] (lysozyme and UDP-N-acetylmuramoyl-l-alanyl-d-glutamate-−2, 6-diaminopimelate ligase), peptidase, ATP synthase and pathogenesis (zonula occludens toxin, Fig. 2c, Table S2).…”

Section: Whole Genome Comparison Between Costa Rican and Apulian Isolatesmentioning

confidence: 99%

Introduction and adaptation of an emerging pathogen to olive trees in Italy

et al. 2021

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The invasive plant pathogen Xylella fastidiosa currently threatens European flora through the loss of economically and culturally important host plants. This emerging vector-borne bacterium, native to the Americas, causes several important diseases in a wide range of plants including crops, ornamentals, and trees. Previously absent from Europe, and considered a quarantine pathogen, X. fastidiosa was first detected in Apulia, Italy in 2013 associated with a devastating disease of olive trees (Olive Quick Decline Syndrome, OQDS). OQDS has led to significant economic, environmental, cultural, as well as political crises. Although the biology of X. fastidiosa diseases have been studied for over a century, there is still no information on the determinants of specificity between bacterial genotypes and host plant species, which is particularly relevant today as X. fastidiosa is expanding in the naive European landscape. We analysed the genomes of 79 X . fastidiosa samples from diseased olive trees across the affected area in Italy as well as genomes of the most genetically closely related strains from Central America. We provided insights into the ecological and evolutionary emergence of this pathogen in Italy. We first showed that the outbreak in Apulia is due to a single introduction from Central America that we estimated to have occurred in 2008 [95 % HPD: 1930–2016]. By using a combination of population genomic approaches and evolutionary genomics methods, we further identified a short list of genes that could play a major role in the adaptation of X. fastidiosa to this new environment. We finally provided experimental evidence for the adaptation of the strain to this new environment.

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Proteomic analysis of Escherichia coli detergent-resistant membranes (DRM)

Cited by 11 publications

References 51 publications

Regulation of the First Committed Step in Lipopolysaccharide Biosynthesis Catalyzed by LpxC Requires the Essential Protein LapC (YejM) and HslVU Protease

Regulation of the First Committed Step in Lipopolysaccharide Biosynthesis Catalyzed by LpxC Requires the Essential Protein LapC (YejM) and HslVU Protease

Escherichia coliSPFH membrane microdomain proteins HflKC contribute to aminoglycoside and oxidative stress tolerance

Introduction and adaptation of an emerging pathogen to olive trees in Italy

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