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Related Experiment Videos

Bacteria lacking a multidrug pump: a sensitive tool for drug discovery

P C Hsieh1, S A Siegel, B Rogers

  • 1Biotechnology Center, Tufts University, 4 Colby Street, Medford, MA 02155, USA.

Proceedings of the National Academy of Sciences of the United States of America
|June 17, 1998
PubMed
Summary

Using multidrug resistance (MDR) pump mutants, like NorA in Staphylococcus aureus, enhances antibiotic discovery. These mutants show significantly increased sensitivity to various antimicrobials, aiding natural product screening.

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Area of Science:

  • Microbiology
  • Biochemistry
  • Pharmacology

Background:

  • Microorganisms utilize multidrug resistance pumps (MDRs) to evade antibiotics, posing challenges for drug discovery.
  • The NorA MDR in Staphylococcus aureus is a key efflux pump contributing to antimicrobial resistance.

Purpose of the Study:

  • To investigate the utility of MDR-deficient mutants for overcoming antibiotic resistance in microbial drug discovery.
  • To evaluate the impact of NorA deficiency and alkaline pH on antimicrobial sensitivity in Staphylococcus aureus.

Main Methods:

  • Generated a Staphylococcus aureus mutant with a disrupted NorA MDR gene.
  • Assessed antimicrobial sensitivity to quinolones and amphipathic cations in wild-type and mutant strains.
  • Investigated the effect of alkaline pH on antimicrobial sensitivity and its synergy with the norA mutation.

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Main Results:

  • The norA mutant exhibited a 5- to 30-fold increase in sensitivity to various antimicrobials.
  • Alkaline pH increased wild-type cell sensitivity by 15- to 60-fold, synergizing with the norA mutation for up to a 1,000-fold increase.
  • The norA- strain showed enhanced sensitivity to plant alkaloid antimicrobials, suggesting NorA's role in their efflux.

Conclusions:

  • MDR-deficient mutants, such as the norA mutant, are valuable tools for enhancing antibiotic discovery and screening.
  • Environmental factors like alkaline pH can significantly modulate antimicrobial sensitivity, particularly in conjunction with MDR mutations.
  • This approach facilitates the identification of novel antimicrobial compounds, including natural products that may be substrates for MDR pumps.