Screening of the PA14NR Transposon Mutant Library Identifies Genes Involved in Resistance to Bacteriophage Infection in Pseudomomas aeruginosa
Name
ijms-25-07009-v2.pdf
Size
4.33 MB
Format
Adobe PDF
Checksum (MD5)
9fd495f951c4204813433be0c26bd992
Author(s)
Ho, Peiying
Dam, Linh Chi
Koh, Wei Ren Ryanna
Nai, Rui Si
Nah, Qian Hui
Rajaie Fizla, Faeqa Binte Muhammad
Chan, Chia Ching
Aung, Thet Tun
Goh, Shin Giek
Fang, You
Date Issued
June 26, 2024
Journal
International Journal of Molecular Sciences
Publisher
MDPI AG
Citation
Ho, P.; Dam, L.C.; Koh, W.R.R.; Nai, R.S.; Nah, Q.H.; Rajaie Fizla, F.B.M.; Chan, C.C.; Aung, T.T.; Goh, S.G.; Fang, Y.; et al. Screening of the PA14NR Transposon Mutant Library Identifies Genes Involved in Resistance to Bacteriophage Infection in Pseudomomas aeruginosa. Int. J. Mol. Sci. 2024, 25, 7009.
Version
Final published version
Abstract
Multidrug-resistant P. aeruginosa infections pose a serious public health threat due to the rise in antimicrobial resistance. Phage therapy has emerged as a promising alternative. However, P. aeruginosa has evolved various mechanisms to thwart phage attacks, making it crucial to decipher these resistance mechanisms to develop effective therapeutic strategies. In this study, we conducted a forward-genetic screen of the P. aeruginosa PA14 non-redundant transposon library (PA14NR) to identify dominant-negative mutants displaying phage-resistant phenotypes. Our screening process revealed 78 mutants capable of thriving in the presence of phages, with 23 of them carrying insertions in genes associated with membrane composition. Six mutants exhibited total resistance to phage infection. Transposon insertions were found in genes known to be linked to phage-resistance such as galU and a glycosyl transferase gene, as well as novel genes such as mexB, lasB, and two hypothetical proteins. Functional experiments demonstrated that these genes played pivotal roles in phage adsorption and biofilm formation, indicating that altering the bacterial membrane composition commonly leads to phage resistance in P. aeruginosa. Importantly, these mutants displayed phenotypic trade-offs, as their resistance to phages inversely affected antibiotic resistance and hindered biofilm formation, shedding light on the complex interplay between phage susceptibility and bacterial fitness. This study highlights the potential of transposon mutant libraries and forward-genetic screens in identifying key genes involved in phage-host interactions and resistance mechanisms. These findings support the development of innovative strategies for combating antibiotic-resistant pathogens.
MIT Department
Singapore-MIT Alliance in Research and Technology (SMART)
Terms of Use
Creative Commons Attribution
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.3390/ijms25137009
