Targets for DNA-based antimicrobials in Pseudomonas aeruginosa

Bahia, Sandeep (2021) Targets for DNA-based antimicrobials in Pseudomonas aeruginosa. Doctoral thesis, University of East Anglia.

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The increasing prevalence of antibiotic resistance is a global issue driving the need for the identification of novel target sites and development of new antimicrobials, in particular for Gram-negative bacteria. Rapid and comprehensive sequencing of microbial pathogens has identified control of bacterial transcription as a source of these needed targets. Transcription factor decoys (TFDs) are short oligonucleotides that competitively inhibit the bacterial transcription factors controlling genes essential for infection. To facilitate the delivery of TFD a delivery molecule, 12-bis-THA, has been developed. An analogue of the quaternary ammonium cation dequalinium, 12-bis-THA has been shown cause membrane disruption in order to delivery TFDs intracellularly. As this disruption results in a stress response likely to be driven by changes in transcription factor (TF) expression, an investigation into detection of these changes using proteomics was explored. In this thesis a proteomic workflow was developed to enrich for the detection of changes in TF expression in response to high and low dose 12-bis-THA treatment, with an aim to identify potential TFD targets. Initial experimentation utilised the model organism E. coli NCTC_9001. Shortcomings highlighted in these experiments informed modification in the workflow for the bacterium of interest, P. aeruginosa. These experiments were successful at identifying changes in TFs expression between treatment conditions. Analysis of the data identified two potential TFD targets, the biofilm resistance locus regulator, BrlR and the alginate and motility regulator, AmrZ. TFDs were then designed to target these TFs and their efficacy explored. The developed proteomic workflow has great potential for application to other bacterial organisms and as a tool for identification of TFD targets and the future development of the novel antimicrobial strategy.

Item Type: Thesis (Doctoral)
Faculty \ School: Faculty of Science > School of Pharmacy
Depositing User: Chris White
Date Deposited: 08 Aug 2022 11:00
Last Modified: 08 Aug 2022 11:00

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