Mycobacterium tuberculosis Rv0191 is an efflux pump of major facilitator superfamily transporter regulated by Rv1353c

ABSTRACT Manogepix is a broad-spectrum antifungal agent that inhibits glycosylphosphatidylinositol (GPI) anchor biosynthesis. Using whole-genome sequencing, we characterized two efflux-mediated mechanisms in the fungal pathogens Candida albicans and Candida parapsilosis that resulted in decreased manogepix susceptibility. In C. albicans, a gain-of-function mutation in the transcription factor gene ZCF29 activated expression of ATP-binding cassette transporter genes CDR11 and SNQ2. In C. parapsilosis, a mitochondrial deletion activated expression of the major facilitator superfamily transporter gene MDR1.

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Efflux Pumps of Mycobacterium tuberculosis Play a Significant Role in Antituberculosis Activity of Potential Drug Candidates

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.06003-11 ◽

2012 ◽

Vol 56 (5) ◽

pp. 2643-2651 ◽

Cited By ~ 87

Author(s):

Meenakshi Balganesh ◽

Neela Dinesh ◽

Sreevalli Sharma ◽

Sanjana Kuruppath ◽

Anju V. Nair ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Efflux Pump ◽

Efflux Pumps ◽

Vital Role ◽

Major Facilitator Superfamily ◽

Content Type ◽

Active Efflux ◽

Drug Candidates ◽

Small Multidrug Resistance ◽

Major Facilitator

ABSTRACTActive efflux of drugs mediated by efflux pumps that confer drug resistance is one of the mechanisms developed by bacteria to counter the adverse effects of antibiotics and chemicals. To understand these efflux mechanisms inMycobacterium tuberculosis, we generated knockout (KO) mutants of four efflux pumps of the pathogen belonging to different classes. We measured the MICs and kill values of two different compound classes on the wild type (WT) and the efflux pump (EP) KO mutants in the presence and absence of the efflux inhibitors verapamil andl-phenylalanyl-l-arginyl-β-naphthylamide (PAβN). Among the pumps studied, the efflux pumps belonging to the ABC (ATP-binding cassette) class, encoded byRv1218c, and the SMR (small multidrug resistance) class, encoded byRv3065, appear to play important roles in mediating the efflux of different chemical classes and antibiotics. Efflux pumps encoded byRv0849andRv1258calso mediate the efflux of these compounds, but to a lesser extent. Increased killing is observed in WTM. tuberculosiscells by these compounds in the presence of either verapamil or PAβN. The efflux pump KO mutants were more susceptible to these compounds in the presence of efflux inhibitors. We have shown that these four efflux pumps ofM. tuberculosisplay a vital role in mediating efflux of different chemical scaffolds. Inhibitors of one or several of these efflux pumps could have a significant impact in the treatment of tuberculosis. The identification and characterization ofRv0849, a new efflux pump belonging to the MFS (major facilitator superfamily) class, are reported.

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An Uncharacterized Major Facilitator Superfamily Transporter From Planococcus maritimus Exhibits Dual Functions as a Na+(Li+, K+)/H+ Antiporter and a Multidrug Efflux Pump

Frontiers in Microbiology ◽

10.3389/fmicb.2018.01601 ◽

2018 ◽

Vol 9 ◽

Cited By ~ 7

Author(s):

Heba Abdel-Motaal ◽

Lin Meng ◽

Zhenglai Zhang ◽

Amro H. Abdelazez ◽

Li Shao ◽

...

Keyword(s):

Efflux Pump ◽

Major Facilitator Superfamily ◽

Multidrug Efflux ◽

Multidrug Efflux Pump ◽

Major Facilitator Superfamily Transporter ◽

Major Facilitator ◽

Dual Functions

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Two Distinct Major Facilitator Superfamily Drug Efflux Pumps Mediate Chloramphenicol Resistance in Streptomyces coelicolor

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00853-09 ◽

2009 ◽

Vol 53 (11) ◽

pp. 4673-4677 ◽

Cited By ~ 24

Author(s):

James J. Vecchione ◽

Blair Alexander ◽

Jason K. Sello

Keyword(s):

Efflux Pump ◽

Streptomyces Coelicolor ◽

Efflux Pumps ◽

Major Facilitator Superfamily ◽

Close Relative ◽

Gram Positive ◽

Antibacterial Drugs ◽

Chloramphenicol Resistance ◽

Drug Activity ◽

Major Facilitator

ABSTRACT Chloramphenicol, florfenicol, and thiamphenicol are used as antibacterial drugs in clinical and veterinary medicine. Two efflux pumps of the major facilitator superfamily encoded by the cmlR1 and cmlR2 genes mediate resistance to these antibiotics in Streptomyces coelicolor, a close relative of Mycobacterium tuberculosis. The transcription of both genes was observed by reverse transcription-PCR. Disruption of cmlR1 decreased the chloramphenicol MIC 1.6-fold, while disruption of cmlR2 lowered the MIC 16-fold. The chloramphenicol MIC of wild-type S. coelicolor decreased fourfold and eightfold in the presence of reserpine and Phe-Arg-β-naphthylamide, respectively. These compounds are known to potentiate the activity of some antibacterial drugs via efflux pump inhibition. While reserpine is known to potentiate drug activity against gram-positive bacteria, this is the first time that Phe-Arg-β-naphthylamide has been shown to potentiate drug activity against a gram-positive bacterium.

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Structural and Functional Study of the Phenicol-Specific Efflux Pump FloR Belonging to the Major Facilitator Superfamily

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.49.7.2965-2971.2005 ◽

2005 ◽

Vol 49 (7) ◽

pp. 2965-2971 ◽

Cited By ~ 27

Author(s):

Martine Braibant ◽

Jacqueline Chevalier ◽

Elisabeth Chaslus-Dancla ◽

Jean-Marie Pagès ◽

Axel Cloeckaert

Keyword(s):

Efflux Pump ◽

Major Facilitator Superfamily ◽

Site Directed Mutagenesis ◽

Functional Study ◽

Efflux Transporters ◽

Chloramphenicol Resistance ◽

Active Efflux ◽

Transmembrane Segments ◽

Efflux Mechanism ◽

Major Facilitator

ABSTRACT The florfenicol-chloramphenicol resistance gene floR from Salmonella enterica was previously identified and postulated to belong to the major facilitator (MF) superfamily of drug exporters. Here, we confirmed a computer-predicted transmembrane topological model of FloR, using the phoA gene fusion method, and classified this protein in the DHA12 family (containing 12 transmembrane domains) of MF efflux transporters. We also showed that FloR is a transporter specific for structurally associated phenicol drugs (chloramphenicol, florfenicol, thiamphenicol) which utilizes the proton motive force to energize an active efflux mechanism. By site-directed mutagenesis of specific charged residues belonging to putative transmembrane segments (TMS), two residues essential for active efflux function, D23 in TMS1 and R109 in TMS4, were identified. Of these, the acidic residue D23 seems to participate directly in the affinity pocket involved in phenicol derivative recognition. A third residue, E283 in TMS9, seems to be necessary for correct membrane folding of the transporter.

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A Major Facilitator Superfamily Transporter Regulated by the Stress-Responsive Transcription Factor Yap1 Is Required for Resistance to Fungicides, Xenobiotics, and Oxidants and Full Virulence in Alternaria alternata

Frontiers in Microbiology ◽

10.3389/fmicb.2018.02229 ◽

2018 ◽

Vol 9 ◽

Cited By ~ 10

Author(s):

Hsien-Che Lin ◽

Pei-Ling Yu ◽

Li-Hung Chen ◽

Hsieh-Chin Tsai ◽

Kuang-Ren Chung

Keyword(s):

Transcription Factor ◽

Alternaria Alternata ◽

Major Facilitator Superfamily ◽

Major Facilitator Superfamily Transporter ◽

Major Facilitator

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The mycobacterial efflux pump EfpA can induce high drug tolerance to many anti-tuberculosis drugs, including moxifloxacin, in Mycobacterium smegmatis

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00262-21 ◽

2021 ◽

Author(s):

Deepika Rai ◽

Sarika Mehra

Keyword(s):

Mycobacterium Smegmatis ◽

Efflux Pump ◽

Drug Tolerance ◽

Inducible Promoter ◽

Major Facilitator Superfamily ◽

Expression Level ◽

Efflux Pump Inhibitor ◽

Gene Encoding ◽

Active Efflux ◽

Major Facilitator

Active efflux of drugs across the membrane is a major survival strategy of bacteria against many drugs. In this work, we characterize an efflux pump EfpA, from the major facilitator superfamily, that is highly conserved among both slow growing and fast-growing mycobacterium species and has been found to be upregulated in many clinical isolates of Mycobacterium tuberculosis . The gene encoding EfpA from Mycobacterium smegmatis was over-expressed under both constitutive and an inducible promoter. Expression of efpA gene under both the promoters resulted in greater than 32-fold increased drug tolerance of M. smegmatis cells to many first-line (rifampicin, isoniazid and streptomycin) and second-line (amikacin) anti-tuberculosis drugs. Notably, drug tolerance of M. smegmatis cells to moxifloxacin increased by more than 180-fold when efpA was over-expressed. The increase in minimum inhibitory concentration (MIC) correlated with the decreased uptake of drugs including norfloxacin, moxifloxacin and ethidium bromide and the high MIC could be reversed in the presence of an efflux pump inhibitor. A correlation was observed between the MIC of drugs and the efflux pump expression level, suggesting that the latter could be modulated by varying the expression level of the efflux pump. The expression of high levels of efpA did not impact the fitness of the cells when supplemented with glucose.The efpA gene is conserved across both pathogenic and non-pathogenic mycobacteria. The efpA gene from the Mycobacterium bovis BCG/ M. tuberculosis , which is 80% identical to efpA from M. smegmatis , also led to decreased antimicrobial efficacy to many drugs, although the fold-change was lower. When over-expressed in M. bovis BCG, an 8-fold higher drug tolerance to moxifloxacin was observed . This is the first report of an efflux pump from mycobacterium species that leads to higher drug tolerance to moxifloxacin, a promising new drug for the treatment of tuberculosis.

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