Skip to main content

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

Publication ,  Journal Article
Xu, F; Hinenoya, A; Zeng, X; Li, X-P; Guan, Z; Lin, J
Published in: Microorganisms
March 22, 2021

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.

Duke Scholars

Published In

Microorganisms

DOI

ISSN

2076-2607

Publication Date

March 22, 2021

Volume

9

Issue

3

Location

Switzerland

Related Subject Headings

  • 3207 Medical microbiology
  • 3107 Microbiology
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Xu, F., Hinenoya, A., Zeng, X., Li, X.-P., Guan, Z., & Lin, J. (2021). Critical Role of 3'-Downstream Region of pmrB in Polymyxin Resistance in Escherichia coli BL21(DE3). Microorganisms, 9(3). https://doi.org/10.3390/microorganisms9030655
Xu, Fuzhou, Atsushi Hinenoya, Ximin Zeng, Xing-Ping Li, Ziqiang Guan, and Jun Lin. “Critical Role of 3'-Downstream Region of pmrB in Polymyxin Resistance in Escherichia coli BL21(DE3).Microorganisms 9, no. 3 (March 22, 2021). https://doi.org/10.3390/microorganisms9030655.
Xu F, Hinenoya A, Zeng X, Li X-P, Guan Z, Lin J. Critical Role of 3'-Downstream Region of pmrB in Polymyxin Resistance in Escherichia coli BL21(DE3). Microorganisms. 2021 Mar 22;9(3).
Xu, Fuzhou, et al. “Critical Role of 3'-Downstream Region of pmrB in Polymyxin Resistance in Escherichia coli BL21(DE3).Microorganisms, vol. 9, no. 3, Mar. 2021. Pubmed, doi:10.3390/microorganisms9030655.
Xu F, Hinenoya A, Zeng X, Li X-P, Guan Z, Lin J. Critical Role of 3'-Downstream Region of pmrB in Polymyxin Resistance in Escherichia coli BL21(DE3). Microorganisms. 2021 Mar 22;9(3).

Published In

Microorganisms

DOI

ISSN

2076-2607

Publication Date

March 22, 2021

Volume

9

Issue

3

Location

Switzerland

Related Subject Headings

  • 3207 Medical microbiology
  • 3107 Microbiology