Regulatory role of RamA and RamR in AcrAB-TolC-mediated carbapenem resistance in carbapenem-resistant hypervirulent Klebsiella pneumoniae
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2026
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Abstract
OBJECTIVE: Carbapenem-resistant hypervirulent Klebsiella pneumoniae (CR-hvKP), which possesses both high pathogenicity and drug resistance, has become a major clinical challenge. The role of AcrAB-TolC efflux pump in its resistance mechanism has attracted increasing attention. This study aimed to clarify the role of the AcrAB-TolC efflux pump in carbapenem resistance of CR-hvKP, confirm the role of AcrB in the function of the AcrAB-TolC efflux pump, and reveal the regulatory mechanism of RamA and RamR. This study provides a theoretical basis for the treatment and prevention of CR-hvKP infections. METHODS: Thirty-eight non-repetitive clinical CR-hvKP isolates were analyzed. Minimum inhibitory concentrations (MICs) were determined using broth microdilution. Efflux pump activity was evaluated by comparing MICs before and after addition of CCCP. The Carbapenemase Nordmann-Poirel (Carb NP) test excluded carbapenemase-mediated resistance. PCR was used to detect virulence (iucA, rmpA, rmpA2, iroB, Peg344) and efflux pump (acrA, acrB, tolC) genes. Bioinformatics tools (KaKs_Calculator and Datamonkey) and RT-qPCR identified and quantified acrB expression. Gene knockout and overexpression, combined with MIC testing and a Galleria mellonella infection model, assessed effects on resistance and virulence. RT-qPCR evaluated RamA and RamR expression. RESULTS: acrB was identified as a major contributor to efflux-mediated resistance. Its expression decreased by 65% in the ramA knockout strain and increased by 40% in the ramR knockout strain. AcrB-positive strains significantly reduced larval survival. CONCLUSION: AcrB appears to be a major component of the AcrAB-TolC efflux system contributing to carbapenem resistance and potentially influencing virulence in CR-hvKP. RamA and RamR significantly regulate AcrB expression and modulate efflux-associated resistance.
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openalex_W7171626772
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| Authors | Ruixuan Yu, Jiaming Tian, Chunrong Hu, Ying Chen, Yong Wang, Shuyue Zhang, Xinran Zhou, Yuehan Gu, Yanjun Fu |
| Journal | Journal of applied microbiology |
| Year | 2026 |
| DOI |
10.1093/jambio/lxag191
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| URL | |
| Keywords | Keywords not found |
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