Overexpression of the flagellar motor protein MotB sensitizes Bacillus subtilis to aminoglycosides in a motility-independent manner

PLoS One. 2024 Apr 26;19(4):e0300634. doi: 10.1371/journal.pone.0300634. eCollection 2024.

Abstract

The flagellar motor proteins, MotA and MotB, form a complex that rotates the flagella by utilizing the proton motive force (PMF) at the bacterial cell membrane. Although PMF affects the susceptibility to aminoglycosides, the effect of flagellar motor proteins on the susceptibility to aminoglycosides has not been investigated. Here, we found that MotB overexpression increased susceptibility to aminoglycosides, such as kanamycin and gentamicin, in Bacillus subtilis without affecting swimming motility. MotB overexpression did not affect susceptibility to ribosome-targeting antibiotics other than aminoglycosides, cell wall-targeting antibiotics, DNA synthesis-inhibiting antibiotics, or antibiotics inhibiting RNA synthesis. Meanwhile, MotB overexpression increased the susceptibility to aminoglycosides even in the motA-deletion mutant, which lacks swimming motility. Overexpression of the MotB mutant protein carrying an amino acid substitution at the proton-binding site (D24A) resulted in the loss of the enhanced aminoglycoside-sensitive phenotype. These results suggested that MotB overexpression sensitizes B. subtilis to aminoglycosides in a motility-independent manner. Notably, the aminoglycoside-sensitive phenotype induced by MotB requires the proton-binding site but not the MotA/MotB complex formation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aminoglycosides* / pharmacology
  • Anti-Bacterial Agents* / pharmacology
  • Bacillus subtilis* / drug effects
  • Bacillus subtilis* / genetics
  • Bacillus subtilis* / metabolism
  • Bacterial Proteins* / genetics
  • Bacterial Proteins* / metabolism
  • Flagella* / drug effects
  • Flagella* / metabolism
  • Molecular Motor Proteins / genetics
  • Molecular Motor Proteins / metabolism

Substances

  • Bacterial Proteins
  • Aminoglycosides
  • MotB protein, Bacteria
  • Anti-Bacterial Agents
  • MotA protein, Bacteria
  • Molecular Motor Proteins

Grants and funding

This study was supported by JSPS Grants-in-Aid for Scientific Research (grants 22K14892, 22H02869, 22K19435, and 23K06130), the Takeda Science Foundation (CK), the Ichiro Kanehara Foundation (CK), the Ryobi Teien Memory Foundation (CK and KI), and Ohmoto Ikueikai Student Grant (RS). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.