Dataset Information

Metabolic reprogramming of Vibrio cholerae impaired in respiratory NADH oxidation is accompanied with increased copper sensitivity

ABSTRACT: The electrogenic, sodium ion translocating NADH:quinone oxidoreductase (NQR) from Vibrio cholerae is frequent in pathogenic bacteria and a potential target for antibiotics. NQR couples the oxidation of NADH to the formation of a sodium motive force (SMF) and therefore drives important processes such as flagellar rotation, substrate uptake, and energy-dissipating cation-proton antiport. We performed a quantitative proteome analysis of V. cholerae O395N1 in comparison to its variant lacking the NQR using minimal medium with glucose as carbon source. We found 84 proteins (≥ regulation factor 2) to be changed in abundance. The loss of NQR resulted in a decrease in abundance of enzymes of the oxidative branch of the TCA cycle and an increase in abundance of virulence factors AcfC and TcpA. Most unexpected, copper resistance proteins CopA, CopG and CueR were decreased in the nqr deletion strain. As a consequence, the mutant exhibited diminished resistance to copper when compared to the reference strain, as confirmed in growth studies using either glucose or mixed amino acids as carbon sources. We propose that the observed adaptations of the nqr deletion strain represent a coordinated response which counteracts a drop in transmembrane voltage that challenges V. cholerae in its different habitats. The dataset also includes raw data and MaxQuant analyses of the treatment of the V. cholerae reference strain and the nqr deletion strain with different catecholamines (WT_B - WT_D and nqr_B - nqr_D). These data will be published in a forthcoming paper.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Vibrio Cholerae O395

TISSUE(S): Endospore, Cell Culture

DISEASE(S): Cholera

SUBMITTER: Jens Pfannstiel

LAB HEAD: Julia Steuber

PROVIDER: PXD009379 | Pride | 2018-06-21

REPOSITORIES: Pride

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Publications

Metabolic Reprogramming of Vibrio cholerae Impaired in Respiratory NADH Oxidation Is Accompanied by Increased Copper Sensitivity.

Toulouse Charlotte C Metesch Kristina K Pfannstiel Jens J Steuber Julia J

Journal of bacteriology 20180710 15

The electrogenic, sodium ion-translocating NADH:quinone oxidoreductase (NQR) from <i>Vibrio cholerae</i> is frequent in pathogenic bacteria and a potential target for antibiotics. NQR couples the oxidation of NADH to the formation of a sodium motive force (SMF) and therefore drives important processes, such as flagellar rotation, substrate uptake, and energy-dissipating cation-proton antiport. We performed a quantitative proteome analysis of <i>V. cholerae</i> O395N1 compared to its variant lack ...[more]

PMID: 29735761

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Project description:The cholera disease bacterium V. cholerae, can adopt planktonic or biofilm lifestyles depending on the intracellular concentration of the second messenger cyclic diguanylic acid (c-di-GMP). Biofilm formation protects Vibrios from stressful conditions and facilitates disease transmission by enhancing infectivity. The histone-like nucleoid structuring protein (H-NS) is a global regulator of genes associated with pathogenicity and responses to environmental stresses. H-NS represses the transcription of genes vpsT, vpsA and vpsL, which are required for the biosynthesis of the biofilm exopolysacchide matrix. Here we demonstrate that the c-di-GMP-binding protein VpsT disrupts H-NS nucleoprotein complexes at the vpsA and vpsL promoters and that this effect is enhanced by c-di-GMP. We used ChIP coupled with Next Generation Sequencing (ChIP-Seq) and transcriptome analysis (RNA-Seq) to identify additional loci repressed by H-NS affecting biofilm formation. This study showed that H-NS directly represses the transcription of genes encoding proteins present in the biofilm matrix such as the rbmA-F cluster, hemolysin and chitinase. Similar to vpsA and vpsL, the promoter region of vpsU, rbmA and rbmF exhibited overlapping H-NS and VpsT binding motifs. Deletion of vpsT increased H-NS occupancy at the vpsU, vpsA, vpsL, rbmA and rbmF promoters. Conversely, artificially increasing the c-di-GMP pool diminished H-NS occupancy at the above promoters. Deletion of vpsT did not affect H-NS occupancy at its own promoter. However, deletion of genes encoding the regulators AphA and VpsR significantly increased H-NS occupancy at the vpsT promoter. In sum, our study shows that c-di-GMP enhances biofilm formation by acting through VpsT to activate an H-NS anti-repression cascade. The Binding profile of V. cholerae H-NS to the genome was determined by ChIP followed by Next Generation Sequencing (ChIP-Seq) using the Illumina HiSeq2000 platform. V. cholerae C7258 cells expressing H-NS-FLAG fusion protein from the hns transcription and translation signals were collected from LB cultures grown to mid-exponential phase (OD600 0.5). An anti-FLAG Immunoprecipitation (IP) and an Input samples were used for the analysis.

			Action	DRS
	ResultsComparisionWTvsdeltaNQR.xlsx	Xlsx
	WT_A1_160312091057.raw	Raw
	WT_A2.raw	Raw
	WT_A3.raw	Raw
	WT_B1.raw	Raw