Transposon Mutagenesis Identifies Novel Genes Associated with Staphylococcus aureus Persister Formation

Frontiers in Microbiology
Wenjie WangYing Zhang

Abstract

Pathogenic bacterial persisters are responsible for the recalcitrance of chronic and persistent infections to antimicrobial therapy. Although the mechanisms of persister formation and survival have been widely studied in Escherichia coli, persistence mechanisms in Staphylococcus aureus remain largely unknown. Here, we screened a transposon mutant library of a clinical methicillin-resistant Staphylococcus aureus(MRSA)strain, USA500 (ST8), under antibiotic pressure and identified 13 genes whose insertion mutations resulted in a defect in persistence. These candidate genes were further confirmed by evaluating the survival of the mutants upon exposure to levofloxacin and several other stress conditions. We found 13 insertion mutants with significantly lower persister numbers under several stress conditions, including sdhA, sdhB, ureG, mnhG1, fbaA, ctaB, clpX, parE, HOU_0223, HOU_0587, HOU_2091, HOU_2315, and HOU_2346, which mapped into pathways of oxidative phosphorylation, TCA cycle, glycolysis, cell cycle, and ABC transporters, suggesting that these genes and pathways may play an important role in persister formation and survival. The newly constructed knockout strains of ureG, sdhA and sdhB and their complemented strains were al...Continue Reading

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Citations

Feb 22, 2018·Antimicrobial Agents and Chemotherapy·Laura Fernández-GarcíaMaría Tomás
Dec 17, 2016·Science·Alexander HarmsKenn Gerdes
Mar 5, 2017·Essays in Biochemistry·Long M G BuiStephen P Kidd
Dec 21, 2019·ACS Synthetic Biology·Heather S DeterNicholas C Butzin

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Methods Mentioned

BETA
PCR
gene knockout

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