Dual Mutations in MSMEG_0965 and MSMEG_1380 Confer High-Level Resistance to Bortezomib and Linezolid by Both Reducing Drug Intake and Increasing Efflux in Mycobacterium smegmatis DOI Open Access
Han Zhang, Cuiting Fang, Buhari Yusuf

et al.

International Journal of Molecular Sciences, Journal Year: 2025, Volume and Issue: 26(8), P. 3779 - 3779

Published: April 17, 2025

The emergence of multidrug-resistant and extensively drug-resistant Mycobacterium tuberculosis strains poses serious challenges to global control, highlighting the urgent need elucidate mechanisms underlying multidrug resistance. In this study, we screened for spontaneous bortezomib (BTZ)-resistant smegmatis (Msm) mutants identified a strain, Msm-R1-2, exhibiting 16- 64-fold increases in minimum inhibitory concentrations (MICs) BTZ linezolid (LZD), respectively, compared parental strain. Whole-genome sequencing revealed resistance-associated mutations two functionally distinct genes: MSMEG_1380, encoding transcriptional regulator involved efflux pump expression, MSMEG_0965, porin protein. CRISPR-Cpf1-assisted gene knockout editing experiments confirmed that single either MSMEG_1380 or MSMEG_0965 caused low-level resistance (4-fold MIC increase) LZD, while dual conferred levels comparable with MICs respectively. An ethidium bromide accumulation assay demonstrated reduce cell wall permeability, contributing Furthermore, quantitative real-time PCR showed upregulate mmpS5-mmpL5 system. Together, these function synergistically: restricted drug entry combined enhanced confers robust These findings provide novel insights into evolutionary mycobacteria.

Language: Английский

Dual Mutations in MSMEG_0965 and MSMEG_1380 Confer High-Level Resistance to Bortezomib and Linezolid by Both Reducing Drug Intake and Increasing Efflux in Mycobacterium smegmatis DOI Open Access
Han Zhang, Cuiting Fang, Buhari Yusuf

et al.

International Journal of Molecular Sciences, Journal Year: 2025, Volume and Issue: 26(8), P. 3779 - 3779

Published: April 17, 2025

The emergence of multidrug-resistant and extensively drug-resistant Mycobacterium tuberculosis strains poses serious challenges to global control, highlighting the urgent need elucidate mechanisms underlying multidrug resistance. In this study, we screened for spontaneous bortezomib (BTZ)-resistant smegmatis (Msm) mutants identified a strain, Msm-R1-2, exhibiting 16- 64-fold increases in minimum inhibitory concentrations (MICs) BTZ linezolid (LZD), respectively, compared parental strain. Whole-genome sequencing revealed resistance-associated mutations two functionally distinct genes: MSMEG_1380, encoding transcriptional regulator involved efflux pump expression, MSMEG_0965, porin protein. CRISPR-Cpf1-assisted gene knockout editing experiments confirmed that single either MSMEG_1380 or MSMEG_0965 caused low-level resistance (4-fold MIC increase) LZD, while dual conferred levels comparable with MICs respectively. An ethidium bromide accumulation assay demonstrated reduce cell wall permeability, contributing Furthermore, quantitative real-time PCR showed upregulate mmpS5-mmpL5 system. Together, these function synergistically: restricted drug entry combined enhanced confers robust These findings provide novel insights into evolutionary mycobacteria.

Language: Английский

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