Dataset Information

ITRAQ dataset of Pseudomonas aeruginosa PAO1 following dially disulfide treatment

ABSTRACT: To establish the effect of DADS on the P. aeruginosa proteome, 50-mL LB medium samples were inoculated at 108 CFU/mL with exponential growth phase P. aeruginosa PAO1. DADS was then added at a concentration of 0 (control) or 0.64 mg/mL, in triplicate. The six experimental groups were incubated for 5 h in a water bath shaker at 37 ºC with a shaking rate of 180 rpm. Cells from the three control groups and three DADS treatment groups were then sampled and centrifuged. The cell precipitate from each experiment group was snap-frozen at -80 ºC.

INSTRUMENT(S): TripleTOF 5600, LTQ

ORGANISM(S): Pseudomonas Aeruginosa Pao1

TISSUE(S): Whole Body

SUBMITTER: Wen-Ru Li

LAB HEAD: Wen-Ru Li

PROVIDER: PXD011144 | Pride | 2019-01-24

REPOSITORIES: Pride

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Publications

Diallyl Disulfide From Garlic Oil Inhibits Pseudomonas aeruginosa Quorum Sensing Systems and Corresponding Virulence Factors.

Li Wen-Ru WR Ma Yong-Kai YK Xie Xiao-Bao XB Shi Qing-Shan QS Wen Xia X Sun Ting-Li TL Peng Hong H

Frontiers in microbiology 20190107

Previously, we determined that diallyl disulfide (DADS) from garlic oil can inhibit Pseudomonas aeruginosa PAO1 pathogenic factors by inactivating the transcription of key genes from three quorum sensing (QS) systems (las, rhl, and pqs) based on the effects of DADS on growth, virulence factor production (elastase, pyocyanin, biofilm, and swarming motility), and RNA transcription (real-time q-PCR). To further investigate the mechanisms underlying the inhibition of the ...[more]

PMID: 30666240

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Project description:Virulence factor production and the development of biofilms in Pseudomonas aeruginosa have been shown to be regulated by two hierarchically organised quorum sensing (QS) systems via small acyl-homoserine lactone (AHL) signal molecules. Recently, a third bacterial signal molecule, the Pseudomonas quinolone signal (PQS), has been identified, which positively regulates a subset of genes dependent on the QS systems. To further dissect the various independent regulation levels for many QS induced genes and to evaluate the impact of PQS on the QS circuitry, we performed a transcriptome analysis of PAO1 cultures supplemented with PQS. The global transcriptional profile in response to PQS revealed a marked up-regulation of genes belonging to the tightly interdependent functional groups of iron acquisition and oxidative stress response. Remarkably, not only most of the differentially regulated genes but also the induction of a lacZ transcriptional fusion of rhlR could be traced back to a iron chelating effect of PQS. Nevertheless, although iron deficiency per se induced rhlR, there seems to be PQS specific effects that are independent of the PQS effect on P. aeruginosa iron homeostasis Experiment Overall Design: For RNA extraction bacteria were harvested at early logarithmic, late logarithmic and stationary phase of growth. Three independent cultures of PQS-treated and untreated PAO1 each were pooled and the RNA was immediately stabilized with RNAprotect Bacteria Reagent (Qiagen, Valencia, CA). RNA was isolated using the RNeasy kit (Qiagen), treated with DNaseI (Roche) for 30 min at 37M-BM-0C and re-purified with the RNeasy spin column. The subsequent steps of cDNA generation and Biotin-ddUTP terminal labelling were performed as described in the manufacturerM-bM-^@M-^Ys instructions for the P. aeruginosa GeneChipM-CM-^R. Fragmented and labelled cDNA was hybridised to a GeneChip at 50 M-BM-0C for 16 h. The washing steps, staining and scanning of the microarrays were performed using the Affymetrix GeneChip system. Data analysis was performed using the Affymetrix Microarray Suite Software 5.0 with Affymetrix default parameters. As expression analysis was performed in duplicate, a total of two GeneChips per culture condition was scanned at 570 nm, 3 M-BM-5m resolution in an Affymetrix GeneChip scanner. The signals were multiplied by a scaling factor to make the average signal for all the arrays equivalent. The data were imported into a Microsoft Access database capable of searching for genes, which were found in all four pairings defined by the Affymetrix Microarray Suite Software as having significant changes in their signal intensities. Data were combined with the latest annotation from the website of the P. aeruginosa PAO1 sequence and the community annotation project provided at www.pseudomonas.com.

Dataset Information

ITRAQ dataset of Pseudomonas aeruginosa PAO1 following dially disulfide treatment

Publications

Diallyl Disulfide From Garlic Oil Inhibits <i>Pseudomonas aeruginosa</i> Quorum Sensing Systems and Corresponding Virulence Factors.

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