University of Sussex
Browse
fmicb-11-575828.pdf (2.17 MB)

Characterization of the mycobacterial MSMEG-3762/63 efflux pump in Mycobacterium smegmatis drug efflux

Download (2.17 MB)
journal contribution
posted on 2023-06-09, 22:19 authored by Barbara De Siena, Nicoletta Campolattano, Gianluca D'Abrosca, Luigi Russo, Daire Cantillon, Rosangela Morasco, Lidia Muscariello, Simon WaddellSimon Waddell, Margherita Sacco
Multi-drug resistant tuberculosis (MDR-TB) represents a major health problem worldwide. Drug efflux and the activity of efflux transporters likely play important roles in the development of drug-tolerant and drug-resistant mycobacterial phenotypes. This study is focused on the action of a mycobacterial efflux pump as a mechanism of drug resistance. Previous studies demonstrated up-regulation of the TetR-like transcriptional regulator MSMEG_3765 in Mycobacterium smegmatis and its ortholog Rv1685c in Mycobacterium tuberculosis (Mtb) in acid-nitrosative stress conditions. MSMEG-3765 regulates the expression of the MSMEG_3762/63/65 operon, and of the orthologous region in Mtb (Rv1687c/86c/85c). MSMEG-3762 and Rv1687c are annotated as ATP-binding proteins, while MSMEG-3763 and Rv1686c are annotated as trans-membrane polypeptides, defining an ABC efflux pump in both M. smegmatis and Mtb. The two putative efflux systems share a high percentage of identity. To examine the role of the putative efflux system MSMEG-3762/63, we constructed and characterized a MSMEG-3763 deletion mutant in M. smegmatis (?MSMEG_3763). By comparative analysis of wild type, knockout, and complemented strains, together with structural modeling and molecular docking bioinformatics analyses of the MSMEG-3763 trans-membrane protein, we define the protein complex MSMEG-3762/63 as an efflux pump. Moreover, we demonstrate involvement of this pump in biofilm development and in the extrusion of rifampicin and ciprofloxacin (CIP), antimicrobial drugs used in first- and second-line anti-TB therapies.

History

Publication status

  • Published

File Version

  • Published version

Journal

Frontiers in Microbiology

ISSN

1664-302X

Publisher

Frontiers Media

Volume

11

Page range

1-11

Article number

a575828

Department affiliated with

  • Global Health and Infection Publications

Full text available

  • Yes

Peer reviewed?

  • Yes

Legacy Posted Date

2020-12-02

First Open Access (FOA) Date

2021-01-11

First Compliant Deposit (FCD) Date

2021-01-11

Usage metrics

    University of Sussex (Publications)

    Categories

    No categories selected

    Licence

    Exports

    RefWorks
    BibTeX
    Ref. manager
    Endnote
    DataCite
    NLM
    DC