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Quantitative proteomics of Pseudomonas protegens Pf-5 treated with pyoluteorin and ΔphlG strain

ABSTRACT: To acquired insights into the pyoluteorin-mediated physiological process of P. fluorescens, label-free quantitative proteomics was used to compare the expression differences between the whole proteome of Pf-5 with and without pyoluteorin. To further investigate the importance of PhlG in physiological processes of P. protegens Pf-5, we performed a quantitative proteomic profiling of wide type and ΔphlG strain.

ORGANISM(S): Pseudomonas Protegens Pf-5

SUBMITTER: Yongxing He

PROVIDER: PXD039317 | iProX | Mon Jan 09 00:00:00 GMT 2023

REPOSITORIES: iProX

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Project description:The GacS/GacA signal transduction system is a central regulator in Pseudomonas spp., including the biological control strain P. fluorescens Pf-5, in which GacS/GacA controls the production of secondary metabolites and exoenzymes that suppress plant pathogens. A whole genome oligonucleotide microarray was developed for Pf-5 and used to assess the global transcriptomic consequences of a gacA mutation in P. fluorescens Pf-5. In cultures at the transition from exponential to stationary growth phase, GacA significantly influenced transcript levels of 632 genes, representing more than 10% of the 6147 annotated genes in the Pf-5 genome. Transcripts of genes involved in the production of hydrogen cyanide, the antibiotic pyoluteorin, and the extracellular protease AprA were at a low level in the gacA mutant, whereas those functioning in siderophore production and other aspects of iron homeostasis were significantly higher in the gacA mutant than in wild-type Pf-5. Notable effects of gacA inactivation were also observed in the transcription of genes encoding components of a type VI secretion system and cytochrome C oxidase subunits. Two novel gene clusters expressed under the control of gacA were identified from transcriptome analysis, and we propose global-regulator-based genome mining as an approach to decipher the secondary metabolome of Pseudomonas spp. In this series two conditions have been analyzed. A gacA mutant of Pseudomonas fluorescens Pf-5 was harvested at early (OD 0.5) and late (OD 2.4) time points and compared to wild-type Pf-5 harvested in parallel. For each slide, an experimental RNA sample from a gacA mutant was labeled with Cy3 or Cy5 and was hybridized with a reference RNA sample from wild-type Pf-5 labeled with the other Cy dye. There are six slides per condition. Each condition is represented by three biological replicates. There are two flip-dye replicates for each biological replicate. Each slide contains three replicate spots per gene.

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Quantitative proteomics of Pseudomonas protegens Pf-5 treated with pyoluteorin and ΔphlG strain

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