scholarly journals Multiple cis-Acting Sequences Mediate Upregulation of the MDR1 Efflux Pump in a Fluconazole-Resistant Clinical Candida albicans Isolate

2006 ◽  
Vol 50 (7) ◽  
pp. 2300-2308 ◽  
Author(s):  
Davina Hiller ◽  
Stephanie Stahl ◽  
Joachim Morschhäuser
Keyword(s):  
Candida Albicans ◽  
Transcription Factors ◽  
Efflux Pump ◽  
Regulatory Region ◽  
Laboratory Strain ◽  
Major Facilitator Superfamily ◽  
Metabolic Inhibitors ◽  
Multidrug Efflux Pump ◽  
Promoter Deletion Analysis ◽  
Major Facilitator

ABSTRACT Overexpression of the MDR1 gene, which encodes a multidrug efflux pump of the major facilitator superfamily, is a frequent cause of resistance to the antimycotic agent fluconazole and other metabolic inhibitors in clinical Candida albicans strains. Constitutive MDR1 overexpression in such strains is caused by mutations in as yet unknown trans-regulatory factors. In order to identify the cis-acting sequences in the MDR1 regulatory region that mediate constitutive MDR1 upregulation, we performed a promoter deletion analysis in the genetic background of an MDR1-overexpressing clinical C. albicans isolate. We found that several different regions in the MDR1 promoter can mediate MDR1 overexpression in this isolate. In contrast, deletion of one of these regions abolished benomyl-induced MDR1 expression in a C. albicans laboratory strain. These results suggest that multiple transcription factors control expression of the MDR1 efflux pump in C. albicans and that the mutation(s) that causes constitutive MDR1 overexpression and drug resistance in clinical C. albicans isolates affects the activities of several of these transcription factors.

scholarly journals Berberine reverses multidrug resistance in Candida albicans by hijacking the drug efflux pump Mdr1p

2020 ◽  
Author(s):  
Yaojun Tong ◽  
Nuo Sun ◽  
Xiangming Wang ◽  
Qi Wei ◽  
Yu Zhang ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Multidrug Resistance ◽  
Efflux Pump ◽  
Efflux Pumps ◽  
Multidrug Resistant ◽  
Major Facilitator Superfamily ◽  
Drug Efflux ◽  
Multidrug Efflux Pump ◽  
Drug Efflux Pumps ◽  
Major Facilitator

AbstractClinical use of antimicrobials faces great challenges from the emergence of multidrug resistant (MDR) pathogens. The overexpression of drug efflux pumps is one of the major contributors to MDR. It is considered as a promising approach to overcome MDR by reversing the function of drug efflux pumps. In the life-threatening fungal pathogen Candida albicans, the major facilitator superfamily (MFS) transporter Mdr1p can excrete many structurally unrelated antifungals, leading to multidrug resistance. Here we report a counterintuitive case of reversing multidrug resistance in C. albicans by using a natural product berberine to hijack the overexpressed Mdr1p for its own importation. Moreover, we illustrate that the imported berberine accumulates in mitochondria, and compromises the mitochondrial function by impairing mitochondrial membrane potential and mitochondrial Complex I. It results in the selective elimination of Mdr1p overexpressed C. albicans cells. Furthermore, we show that berberine treatment can prolong the mean survival time (MST) of mice with a blood-borne dissemination of Mdr1p overexpressed multidrug resistant candidiasis. This study provided a potential direction of novel anti-MDR drug discovery by screening for multidrug efflux pump converters.

scholarly journals Differential Requirement of the Transcription Factor Mcm1 for Activation of the Candida albicans Multidrug Efflux PumpMDR1by Its Regulators Mrr1 and Cap1

2011 ◽  
Vol 55 (5) ◽  
pp. 2061-2066 ◽  
Author(s):  
Selene Mogavero ◽  
Arianna Tavanti ◽  
Sonia Senesi ◽  
P. David Rogers ◽  
Joachim Morschhäuser
Keyword(s):  
Transcription Factor ◽  
Candida Albicans ◽  
Transcription Factors ◽  
Molecular Mechanisms ◽  
Efflux Pump ◽  
Multidrug Efflux ◽  
Multidrug Efflux Pump ◽  
Gain Of Function ◽  
Pathogenic Yeast ◽  
Content Type

ABSTRACTOverexpression of the multidrug efflux pump Mdr1 causes increased fluconazole resistance in the pathogenic yeastCandida albicans. The transcription factors Mrr1 and Cap1 mediateMDR1upregulation in response to inducing stimuli, and gain-of-function mutations in Mrr1 or Cap1, which render the transcription factors hyperactive, result in constitutiveMDR1overexpression. The essential MADS box transcription factor Mcm1 also binds to theMDR1promoter, but its role in inducible or constitutiveMDR1upregulation is unknown. Using a conditional mutant in which Mcm1 can be depleted from the cells, we investigated the importance of Mcm1 forMDR1expression. We found that Mcm1 was dispensable forMDR1upregulation by H2O2but was required for fullMDR1induction by benomyl. A C-terminally truncated, hyperactive Cap1 could upregulateMDR1expression both in the presence and in the absence of Mcm1. In contrast, a hyperactive Mrr1 containing a gain-of-function mutation depended on Mcm1 to causeMDR1overexpression. These results demonstrate a differential requirement for the coregulator Mcm1 for Cap1- and Mrr1-mediatedMDR1upregulation. When activated by oxidative stress or a gain-of-function mutation, Cap1 can induceMDR1expression independently of Mcm1, whereas Mrr1 requires either Mcm1 or an active Cap1 to cause overexpression of theMDR1efflux pump. Our findings provide more detailed insight into the molecular mechanisms of drug resistance in this important human fungal pathogen.

scholarly journals Candida albicans Zn Cluster Transcription Factors Tac1 and Znc1 Are Activated by Farnesol To Upregulate a Transcriptional Program Including the Multidrug Efflux Pump CDR1

2018 ◽  
Vol 62 (11) ◽  
Author(s):  
Zhongle Liu ◽  
John M. Rossi ◽  
Lawrence C. Myers
Keyword(s):  
Candida Albicans ◽  
Transcription Factors ◽  
Quorum Sensing ◽  
Efflux Pump ◽  
Close Relative ◽  
Multidrug Efflux ◽  
Multidrug Efflux Pump ◽  
Content Type ◽  
Dna Binding Specificity ◽  
Hyphal Formation

ABSTRACT Farnesol, a quorum-sensing molecule, inhibits Candida albicans hyphal formation, affects its biofilm formation and dispersal, and impacts its stress response. Several aspects of farnesol's mechanism of action remain incompletely uncharacterized. Among these are a thorough accounting of the cellular receptors and transporters for farnesol. This work suggests these processes are linked through the Zn cluster transcription factors Tac1 and Znc1 and their induction of the multidrug efflux pump Cdr1. Specifically, we have demonstrated that Tac1 and Znc1 are functionally activated by farnesol through a mechanism that mimics other means of hyperactivation of Zn cluster transcription factors. This is consistent with our observation that many genes acutely induced by farnesol are dependent on TAC1, ZNC1, or both. A related molecule, 1-dodecanol, invokes a similar TAC1-ZNC1 response, while several other proposed C. albicans quorum-sensing molecules do not. Tac1 and Znc1 both bind to and upregulate the CDR1 promoter in response to farnesol. Differences in inducer and DNA binding specificity lead to Tac1 and Znc1 having overlapping, but nonidentical, regulons. Induction of genes by farnesol via Tac1 and Znc1 was inversely related to the level of CDR1 present in the cell, suggesting a model in which induction of CDR1 by Tac1 and Znc1 leads to an increase in farnesol efflux. Consistent with this premise, our results show that CDR1 expression, and its regulation by TAC1 and ZNC1, facilitates growth in the presence of high farnesol concentrations in C. albicans and in certain strains of its close relative, C. dubliniensis.

scholarly journals Inducible and Constitutive Activation of Two Polymorphic Promoter Alleles of the Candida albicans Multidrug Efflux PumpMDR1

2012 ◽  
Vol 56 (8) ◽  
pp. 4490-4494 ◽  
Author(s):  
Christoph Sasse ◽  
Rebecca Schillig ◽  
Alexandra Reimund ◽  
Julia Merk ◽  
Joachim Morschhäuser
Keyword(s):  
Candida Albicans ◽  
Transcription Factors ◽  
Loss Of Heterozygosity ◽  
Efflux Pump ◽  
Multidrug Efflux ◽  
Multidrug Efflux Pump ◽  
Fluconazole Resistance ◽  
Content Type ◽  
Constitutive Activation ◽  
Strain Sc5314

ABSTRACTOverexpression of the multidrug efflux pumpMDR1confers resistance to the antifungal drug fluconazole onCandida albicans. It has been reported that two types ofMDR1promoters exist inC. albicansand that homozygosity for the allele with higher activity may promote fluconazole resistance. We found that the twoMDR1promoter alleles in strain SC5314 were equally well activated by inducing chemicals or hyperactive forms of the transcription factors Mrr1 and Cap1, which controlMDR1expression. In addition, no loss of heterozygosity at theMDR1locus was observed inMDR1-overexpressing clinicalC. albicansstrains that developed fluconazole resistance during therapy.

scholarly journals Involvement of the SmeAB Multidrug Efflux Pump in Resistance to Plant Antimicrobials and Contribution to Nodulation Competitiveness in Sinorhizobium meliloti

2011 ◽  
Vol 77 (9) ◽  
pp. 2855-2862 ◽  
Author(s):  
Shima Eda ◽  
Hisayuki Mitsui ◽  
Kiwamu Minamisawa
Keyword(s):  
Antimicrobial Resistance ◽  
Sinorhizobium Meliloti ◽  
Efflux Pump ◽  
Major Facilitator Superfamily ◽  
Multidrug Efflux ◽  
Multidrug Efflux Pump ◽  
Content Type ◽  
Multidrug Efflux Pumps ◽  
Nodulation Competitiveness ◽  
Major Facilitator

ABSTRACTThe contributions of multicomponent-type multidrug efflux pumps to antimicrobial resistance and nodulation ability inSinorhizobium melilotiwere comprehensively analyzed. Computational searches identified genes in theS. melilotistrain 1021 genome encoding 1 pump from the ATP-binding cassette family, 3 pumps from the major facilitator superfamily, and 10 pumps from the resistance-nodulation-cell division family, and subsequently, these genes were deleted either individually or simultaneously. Antimicrobial susceptibility tests demonstrated that deletion of thesmeABpump genes resulted in increased susceptibility to a range of antibiotics, dyes, detergents, and plant-derived compounds and, further, that specific deletion of thesmeCDorsmeEFgenes in a ΔsmeABbackground caused a further increase in susceptibility to certain antibiotics. Competitive nodulation experiments revealed that thesmeABmutant was defective in competing with the wild-type strain for nodulation. The introduction of a plasmid carryingsmeABinto thesmeABmutant restored antimicrobial resistance and nodulation competitiveness. These findings suggest that the SmeAB pump, which is a major multidrug efflux system ofS. meliloti, plays an important role in nodulation competitiveness by mediating resistance toward antimicrobial compounds produced by the host plant.

scholarly journals LmrS Is a Multidrug Efflux Pump of the Major Facilitator Superfamily from Staphylococcus aureus

2010 ◽  
Vol 54 (12) ◽  
pp. 5406-5412 ◽  
Author(s):  
Jody L. Floyd ◽  
Kenneth P. Smith ◽  
Sanath H. Kumar ◽  
Jared T. Floyd ◽  
Manuel F. Varela
Keyword(s):  
Staphylococcus Aureus ◽  
Efflux Pump ◽  
Sodium Dodecyl ◽  
Fold Increase ◽  
Relative Increase ◽  
Major Facilitator Superfamily ◽  
Multidrug Efflux ◽  
Multidrug Efflux Pump ◽  
Major Facilitator ◽  
Lincomycin Resistance

ABSTRACT A multidrug efflux pump designated LmrS (lincomycin resistance protein of Staphylococcus aureus), belonging to the major facilitator superfamily (MFS) of transporters, was cloned, and the role of LmrS in antimicrobial efflux was evaluated. The highest relative increase in MIC, 16-fold, was observed for linezolid and tetraphenylphosphonium chloride (TPCL), followed by an 8-fold increase for sodium dodecyl sulfate (SDS), trimethoprim, and chloramphenicol. LmrS has 14 predicted membrane-spanning domains and is homologous to putative lincomycin resistance proteins of Bacillus spp., Lactobacillus spp., and Listeria spp.

scholarly journals Structure of the periplasmic adaptor protein from a major facilitator superfamily (MFS) multidrug efflux pump

FEBS Letters ◽  
2014 ◽  
Vol 588 (17) ◽  
pp. 3147-3153 ◽  
Author(s):  
Philip Hinchliffe ◽  
Nicholas P. Greene ◽  
Neil G. Paterson ◽  
Allister Crow ◽  
Colin Hughes ◽  
...  
Keyword(s):  
Efflux Pump ◽  
Adaptor Protein ◽  
Major Facilitator Superfamily ◽  
Multidrug Efflux ◽  
Multidrug Efflux Pump ◽  
Major Facilitator

scholarly journals Overexpression of the MDR1 Gene Is Sufficient To Confer Increased Resistance to Toxic Compounds in Candida albicans

2006 ◽  
Vol 50 (4) ◽  
pp. 1365-1371 ◽  
Author(s):  
Davina Hiller ◽  
Dominique Sanglard ◽  
Joachim Morschhäuser
Keyword(s):  
Drug Resistance ◽  
Candida Albicans ◽  
Efflux Pump ◽  
Brefeldin A ◽  
Major Facilitator Superfamily ◽  
Toxic Compounds ◽  
Human Fungal Pathogen ◽  
Fluconazole Resistant ◽  
Drug Resistant Phenotype ◽  
Major Facilitator

ABSTRACT Overexpression of MDR1, which encodes a membrane transport protein of the major facilitator superfamily, is one mechanism by which the human fungal pathogen Candida albicans can develop increased resistance to the antifungal drug fluconazole and other toxic compounds. In clinical C. albicans isolates, constitutive MDR1 overexpression is accompanied by the upregulation of other genes, but it is not known if these additional alterations are required for Mdr1p function and drug resistance. To investigate whether MDR1 overexpression is sufficient to confer a drug-resistant phenotype in C. albicans, we expressed the MDR1 gene from the strong ADH1 promoter in C. albicans laboratory strains that did not express the endogenous MDR1 gene as well as in a fluconazole-resistant clinical C. albicans isolate in which the endogenous MDR1 alleles had been deleted and in a matched fluconazole-susceptible isolate from the same patient. Forced MDR1 overexpression resulted in increased resistance to the putative Mdr1p substrates cerulenin and brefeldin A, and this resistance did not depend on the additional alterations which occurred during drug resistance development in the clinical isolates. In contrast, artificial expression of the MDR1 gene from the ADH1 promoter did not enhance or only slightly enhanced fluconazole resistance, presumably because Mdr1p expression levels in the transformants were considerably lower than those observed in the fluconazole-resistant clinical isolate. These results demonstrate that MDR1 overexpression in C. albicans is sufficient to confer resistance to some toxic compounds that are substrates of this efflux pump but that the degree of resistance depends on the Mdr1p expression level.

scholarly journals The Quorum-Sensing Molecule Farnesol Is a Modulator of Drug Efflux Mediated by ABC Multidrug Transporters and Synergizes with Drugs in Candida albicans

2011 ◽  
Vol 55 (10) ◽  
pp. 4834-4843 ◽  
Author(s):  
Monika Sharma ◽  
Rajendra Prasad
Keyword(s):  
Candida Albicans ◽  
Quorum Sensing ◽  
Efflux Pump ◽  
Major Facilitator Superfamily ◽  
Simultaneous Increase ◽  
Drug Efflux ◽  
Multidrug Efflux Pump ◽  
Content Type ◽  
Multidrug Transporters ◽  
Quorum Sensing Molecule

ABSTRACTOverexpression of theCaCDR1-encoded multidrug efflux pump protein CaCdr1p (Candidadrug resistance protein 1), belonging to the ATP binding cassette (ABC) superfamily of transporters, is one of the most prominent contributors of multidrug resistance (MDR) inCandida albicans. Thus, blocking or modulating the function of the drug efflux pumps represents an attractive approach in combating MDR. In the present study, we provide first evidence that the quorum-sensing molecule farnesol (FAR) is a specific modulator of efflux mediated by ABC multidrug transporters, such as CaCdr1p and CaCdr2p ofC. albicansand ScPdr5p ofSaccharomyces cerevisiae. Interestingly, FAR did not modulate the efflux mediated by the multidrug extrusion pump protein CaMdr1p, belonging to the major facilitator superfamily (MFS). Kinetic data revealed that FAR competitively inhibited rhodamine 6G efflux in CaCdr1p-overexpressing cells, with a simultaneous increase in an apparentKmwithout affecting theVmaxvalues and the ATPase activity. We also observed that when used in combination, FAR at a nontoxic concentration synergized with the drugs at their respective nonlethal concentrations, as was evident from their <0.5 fractional inhibitory concentration index (FICI) values and from the drop of 14- to 64-fold in the MIC80values in the wild-type strain and in azole-resistant clinical isolates ofC. albicans. Our biochemical experiments revealed that the synergistic interaction of FAR with the drugs led to reactive oxygen species accumulation, which triggered early apoptosis, and that both could be partly reversed by the addition of an antioxidant. Collectively, FAR modulates drug extrusion mediated exclusively by ABC proteins and is synergistic to fluconazole (FLC), ketoconazole (KTC), miconazole (MCZ), and amphotericin (AMB).

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