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Critical Role of 3 '-Downstream Region of pmrB in Polymyxin Resistance in Escherichia coli BL21(DE3)

文献类型： 外文期刊

作者： Xu, Fuzhou 1 ; Hinenoya, Atsushi 1 ; Zeng, Ximin 1 ; Li, Xing-Ping 1 ; Guan, Ziqiang 6 ; Lin, Jun 1 ;

作者机构： 1.Univ Tennessee, Dept Anim Sci, Knoxville, TN 37996 USA

2.Beijing Acad Agr & Forestry Sci, Inst Anim Husb & Vet Med, Beijing 100097, Peoples R China

3.Osaka Prefecture Univ, Grad Sch Life & Environm Sci, Osaka 5998531, Japan

4.Osaka Prefecture Univ, Asian Hlth Sci Inst, Osaka 5998531, Japan

5.Henan Univ Sci & Technol, Coll Anim Sci & Technol, Luoyang 471000, Peoples R China

6.Duke Univ, Med Ctr, Dept Biochem, Durham, NC 27708 USA

关键词： polymyxin resistance; two-component regulatory system; mRNA decay; lipid A modification

期刊名称：MICROORGANISMS （ 影响因子：4.152； ）

ISSN：

年卷期： 2021 年 9 卷 3 期

页码：

收录情况： SCI

摘要： Polymyxins, such as colistin and polymyxin B, are the drugs used as a last resort to treat multidrug-resistant Gram-negative bacterial infections in humans. Increasing colistin resistance has posed a serious threat to human health, warranting in-depth mechanistic research. In this study, using a functional cloning approach, we examined the molecular basis of colistin resistance in Escherichia coli BL21(DE3). Five transformants with inserts ranging from 3.8 to 10.7 kb displayed significantly increased colistin resistance, three of which containing pmrB locus and two containing pmrD locus. Stepwise subcloning indicated that both the pmrB with a single G361A mutation and at least a 103 bp downstream region of pmrB are essential for conferring colistin resistance. Analysis of the mRNA level and stability showed that the length of the downstream region drastically affected the pmrB mRNA level but not its half-life. Lipid A analysis, by mass spectrometry, revealed that the constructs containing pmrB with a longer downstream region (103 or 126 bp) have charge-altering l-4-aminoarabinose (Ara4N) and phosphoethanolamine (pEtN) modifications in lipid A, which were not observed in both vector control and the construct containing pmrB with an 86 bp downstream region. Together, the findings from this study indicate that the 3 '-downstream region of pmrB is critical for the PmrB-mediated lipid A modifications and colistin resistance in E. coli BL21(DE3), suggesting a novel regulatory mechanism of PmrB-mediated colistin resistance in E. coli.

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