Dataset Information

Proteomic analyses of Pseudomonas infected with giant phage PhiKZ

ABSTRACT: Preliminary analyses of Pseudomonas aeruginosa cells infected with phage PhiKZ and a control at 20 minutes post infection.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Pseudomonas Phage Phikz

SUBMITTER: Julie Thomas

LAB HEAD: Julie Thomas

PROVIDER: PXD011520 | Pride | 2018-11-21

REPOSITORIES: Pride

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Publications

Global Proteomic Profiling of Salmonella Infection by a Giant Phage.

Weintraub Susan T ST Mohd Redzuan Nurul Humaira NH Barton Melissa K MK Md Amin Nur Amira NA Desmond Maxim I MI Adams Lily E LE Ali Bazla B Pardo Sammy S Molleur Dana D Wu Weimin W Newcomb William W WW Osier Michael V MV Black Lindsay W LW Steven Alasdair C AC Thomas Julie A JA

Journal of virology 20190219 5

The 240-kb Salmonella phage SPN3US genome encodes 264 gene products, many of which are functionally uncharacterized. We have previously used mass spectrometry to define the proteomes of wild-type and mutant forms of the SPN3US virion. In this study, we sought to determine whether this technique was suitable for the characterization of the SPN3US proteome during liquid infection. Mass spectrometry of SPN3US-infected cells identified 232 SPN3US and 1,994 Salmonella proteins. SPN3US p ...[more]

PMID: 30541839

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Project description:Biofilms have been implicated in delayed wound healing, although the mechanisms by which biofilms impair wound healing are poorly understood. Many species of bacteria produce exotoxins and exoenzymes that may inhibit healing. In addition, oxygen consumption by biofilms, as well as responding leukocytes, may impede wound healing. In this study, we used oxygen microsensors to measure oxygen transects through in vitro-cultured biofilms, biofilms formed in vivo within scabs from a diabetic (db/db) mouse model, and ex vivo human chronic wound specimens. The results show that oxygen levels within mouse scabs had steep gradients that reached minima ranging from 17-72 mmHg on live mice and 6.4-1.1 mmHg on euthanized mice. The oxygen gradients in the mouse scabs were similar to those observed for clinical isolates cultured in vitro and for human ex vivo specimens. No oxygen gradients were observed for heat-killed mouse scabs, suggesting that active metabolism by the viable bacteria and host cells contributed to the reduced oxygen partial pressure of the scabs. To characterize the metabolic activities of the bacteria in the mouse scabs, we performed transcriptomics analyses of Pseudomonas aeruginosa biofilms associated with the db/db mice wounds using Affymetrix microarrays. The results demonstrated that the bacteria expressed genes for metabolic activities associated with cell growth. Interestingly, the transcriptome results indicated that the bacteria within the wounds also experienced oxygen-limitation stress. Among the bacterial genes that were expressed in vivo were genes associated with the Anr-mediated hypoxia-stress response. Other bacterial stress response genes highly expressed in vivo were genes associated with stationary-phase growth, osmotic stress, and RpoH-mediated heat shock stress. Overall, the results support the hypothesis that bacterial biofilms in chronic wounds promote chronicity by contributing to the maintenance of localized low oxygen tensions. Transcriptional profiling of two independent biological replicates of Pseudomonas aeruginosa biofilms, as grown to 72 hours and used as inocula applied to the murine wounds, was performed. A principle components analysis (PCA) was used to provide an overview of the transcriptome data from the 28-day mouse wound scab, comparing the data to the biofilm inoculum, and to published reports of P. aeruginosa biofilm and planktonic samples. The analysis shows that the transcriptome of the mouse wound scab was distinct from the biofilm inoculum that was applied to the wound, demonstrating a shift in biofilm gene expression following 28 days of infection. We sought to characterize P. aeruginosa activity within biofilms in the mouse wound model by isolating and identifying mRNA from the biofilms used as inocula and from the wound scabs 28 days post infection.

Dataset Information

Proteomic analyses of Pseudomonas infected with giant phage PhiKZ

Publications

Global Proteomic Profiling of <i>Salmonella</i> Infection by a Giant Phage.

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