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P. aeruginosa/S. aureus wt, fhuG mutant, fhuD1/D2 double mutant interaction

ABSTRACT: Manuscript Title: Pyochelin biotransformation shapes bacterial competition LC-MS/MS analysis of P. aeruginosa interaction with S. aureus wildtype, fhuG mutant and fhuD1/D2 double mutant.

INSTRUMENT(S): impact HD

ORGANISM(S): Staphylococcus Aureus Lac (ncbitaxon:867914) Pseudomonas Aeruginosa Pao1 (ncbitaxon:208964)

SUBMITTER: Christian Jenul

PROVIDER: MSV000089231 | MassIVE | Sun Apr 10 18:32:00 BST 2022

REPOSITORIES: MassIVE

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Project description:Transcriptional profiles of Escherichia coli MG1655 in mixed culture with Pseudomonas aeruginosa PAO1 showed a number of E. coli genes to be upregulated including purA-F and other genes associated with purine synthesis. In contrast, genes associated with pyrimidine synthesis were unaffected. Competition experiments in both planktonic and biofilm cultures, using three purine synthesis mutants, purD, purH, and purT showed little difference in E. coli survival from the parent strain. As purines are components of the cell signals, cAMP and c-di-GMP, we conducted competition experiments with E. coli mutants lacking adenylate cyclase (cyaA), cAMP phosphodiesterase (cpdA), and the catabolite receptor protein (crp), as well as ydeH, an uncharacterized gene that has been associated with c-di-GMP synthesis. Survival of the cyaA and crp mutants during co-culture were significantly less than the parent strain. Supplementation of the media with 1mM cAMP could restore survival of the cyaA mutant but not the crp mutant. In contrast, survival of the cpdA mutant was similar to the parent strain. Survival of the ydeH mutant was moderately less than the parent, suggesting that cAMP has more impact on E. coli mixed culture growth than c-di-GMP. Addition of 1 mM indole restored the survival of both the cyaA and crp mutations. Mutants in genes for tryptophan synthesis (trpE) and indole production (tnaA) showed a loss of competition and recovery through indole supplementation, comparable to the cyaA and crp mutants. Overall, these results suggest indole and cAMP as major contributing factors to E. coli growth in mixed culture. Two replicates of LB grown E. coli ZK126 and P. aeruginosa PAO1 were grown in pure culture at 37C to an OD of 0.3 (log phase) and then mixed 1:1 and incubated for an additional 45 min. RNA was extracted, purified, reverse transcribed to cDNA and then analyzed on E. coli and P. aeruginosa chips from Affymetrix. Expression profiles were compared to pure culture E. coli and P. aeruginosa grown on LB.

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P. aeruginosa/S. aureus wt, fhuG mutant, fhuD1/D2 double mutant interaction

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