PDR3, a new yeast regulatory gene, is homologous toPDR1 and controls the multidrug resistance phenomenon

Delaveau, Thierry; Delahodde, Agnès; Carvajal, Elvira; Šubík, Július; Jacq, Claude

doi:10.1007/bf00583901

Cited by 203 publications

(194 citation statements)

References 56 publications

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“…5, Pdr1p more strongly influences oligomycin resistance than its homolog Pdr3p (compare the FY1679/EC (pdr1⌬) and FY1679/TD (pdr3⌬) strains). This is in agreement with previous observations (12) even though no difference in oligomycin resistance between pdr1⌬ and pdr3⌬ strains was observed by Delaveau et al (44). As also shown by Fig.…”

Section: Resultssupporting

confidence: 94%

ATPase and Multidrug Transport Activities of the Overexpressed Yeast ABC Protein Yor1p

Decottignies¹,

Grant²,

Nichols³

et al. 1998

Journal of Biological Chemistry

363

334

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The Saccharomyces cerevisiae genome encodes 15 fullsize ATP binding cassette transporters (ABC), of which PDR5, SNQ2, and YOR1 are known to be regulated by the transcription factors Pdr1p and Pdr3p (pleiotropic drug resistance). We have identified two new ABC transporter-encoding genes, PDR10 and PDR15, which were upregulated by the PDR1-3 mutation. These genes, as well as four other ABC transporter-encoding genes, were deleted in order to study the properties of Yor1p. The PDR1-3 gain-of-function mutant was then used to overproduce Yor1p up to 10% of the total plasma membrane proteins. Despite their different topologies, both Yor1p and Pdr5p mediated the ATP-dependent translocation of similar drugs and phospholipids across the yeast cell membrane. Both ABC transporters exhibit ATP hydrolysis in vitro, but Pdr5p ATPase activity is about 15 times higher than that of Yor1p, which may indicate mechanistic or regulatory differences between the two enzymes. The yeast YOR11 gene confers oligomycin resistance on overexpression in a 2-m plasmid (1). Its nucleotide sequence reveals an ORF of 1477 amino acids encoding an ABC protein highly homologous to mammalian transporters such as the multidrug resistance-conferring enzyme MRP (BLAST (see Ref. 2) sequence homology score: p ϭ e Ϫ228 ), the organic anion transporter cMOAT (p ϭ e Ϫ216 ), the sulfonylurea receptor (p ϭ e Ϫ164 ), and the cystic fibrosis transmembrane conductance regulator CFTR (p ϭ e Ϫ132 ). Yor1p is a "full-size" ABC transporter with the topology (TM-NBF) 2 (3, 4). It consists of two homologous halves, with each containing a putative ATP-binding domain (NBF) and a transmembrane domain of six membrane spans (TM). Cui et al. (5) showed that Yor1p confers resistance to a series of drugs including reveromycin A and suggested that Yor1p may be involved in the cellular efflux of organic anions including the fluorescent dye rhodamine B. They also showed that incubation with reveromycin A increases the YOR1 mRNA level. The transcription of YOR1 is controlled by the homologous pair of transcription factors Pdr1p/Pdr3p. The level of YOR1 transcription is decreased by the deletion of either PDR1 or PDR3 and increased in the presence of the gain-of-function PDR1 alleles (1).In this paper, we have investigated the transport activity of Yor1p. Building on previous studies, which indicated that the (TM-NBF) 2 -type Yor1p, together with the (NBF-TM) 2 -type Pdr5p and Snq2p ABC transporters, are overexpressed in the PDR1-3 mutant plasma membrane (6 -8), the PDR1-3 mutant has been used as a tool that enhances the Yor1p protein level. As another investigative tool, we constructed a set of isogenic strains, in the PDR1-3 mutant, with multiple deletions of homologous ABC genes since, in situations where two or more proteins located in the same subcellular compartment share a common substrate, a clear phenotype is only seen when all the corresponding genes are deleted, as illustrated by the work of Mahé et al. (9), who showed that Pdr5p and Snq2p have an overlapping transport ...

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Section: Resultssupporting

confidence: 94%

ATPase and Multidrug Transport Activities of the Overexpressed Yeast ABC Protein Yor1p

Decottignies¹,

Grant²,

Nichols³

et al. 1998

Journal of Biological Chemistry

363

334

View full text Add to dashboard Cite

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“…The socalled MHR is found in a large subset of the family but not War1 (43). Notably, mutations within this putative MHR cause hyperactivity of Leu3 and Pdr1/Pdr3 (16,42,44). Likewise, our data demonstrate that loss of the central region leads to constitutive War1 activity.…”

Section: Conserved Internal Domains Have Inhibitory Functions-mentioning

confidence: 57%

“…One highly interesting allele is truncated, encoding a translational stop instead of the conserved tryptophan residue 936. Truncation of Put3 and Pdr1/Pdr3 also leads to constitutively active forms (41,42). A similar effect was achieved by introducing a hemagglutinin tag at the very C terminus of Put3 (36), and a tandem affinity purification tag in the Pdr3 regulator.…”

Section: Conserved Internal Domains Have Inhibitory Functions-mentioning

confidence: 86%

Weak Organic Acids Trigger Conformational Changes of the Yeast Transcription Factor War1 in Vivo to Elicit Stress Adaptation

Gregori

Schüller

Frohner

et al. 2008

Journal of Biological Chemistry

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The Saccharomyces cerevisiae zinc cluster regulator War1 mediates an essential transcriptional and adaptive response to weak organic acid stress. Here we investigate the mechanism of War1 activation upon weak acid stress. We identified several gain-of-function WAR1 alleles mapping to the central War1 region. These mutations constitutively increase levels of the plasma membrane ABC transporter Pdr12, the main War1 target mediating stress adaptation. Functional analysis of War1 reveals that the central region and its C-terminal activation domain are required for function. Notably, the native DNAbinding and dimerization domains appear dispensable for War1 activity, because they can be replaced by a LexA DNA-binding domain. Chromatin immunoprecipitation demonstrates elevated promoter affinity of activated War1, because its PDR12 promoter association increases upon stress. Hyperactive WAR1 alleles have constitutively high PDR12 promoter association. Furthermore, fluorescence resonance energy transfer of functional CFP-War1-YFP proteins also demonstrates conformational changes of stress-activated War1 in vivo. Our results suggest a mechanism whereby War1 activation is accompanied by conformational changes enhancing promoter association, thus initiating the adaptation process.Weak organic acids, such as potassium sorbate, calcium propionate, and sodium benzoate, are commonly used to preserve foods and beverages (1). In Saccharomyces cerevisiae, uncharged weak acids diffuse across the plasma membrane and dissociate inside the cell, causing intracellular acidification, free radical production, and oxidative stress (1), as well as inhibition of several metabolic processes (2). The yeast ABC transporter Pdr12 confers resistance to monocarboxylic acids with chain lengths up to C 7 (3, 4). Only organic acids but not other stresses cause a dramatic induction of PDR12 transcription and a concomitant increase of Pdr12 (4, 5). A physiological role of Pdr12 is suggested from its proposed ability to extrude amino acid catabolism by-products (6). Weak acid regulation of PDR12 occurs at the level of transcription and requires the Zn(II) 2 Cys 6 zinc cluster transcription factor War1, which recognizes and binds response elements within the PDR12 promoter (7). War1 is functionally conserved among several yeast species, including fungal pathogens. An orthologue mediating sorbate tolerance was identified in the distantly related human fungal pathogen Candida albicans (8), but the mechanism by which weak organic acids cause War1 activation is currently an open question.In S. cerevisiae, some 55 members of the binuclear zinc cluster regulator family exist (for a review see Ref. 9). Most exhibit a similar molecular architecture and domain organization. The N-terminal DNA-binding domain is followed by a coiled-coil dimerization domain, by an extended region of limited homology referred to as the "middle homology region" (MHR), 3 and finally by a short acidic stretch encompassing the C-terminal activation domain (10). Fungal zinc cluster pro...

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“…[17][18][19][20][21][22][23][24][25][26][27][28] C. albicans Fcr1p displays significant sequence homology with S. cerevisiae Pdr1p and Pdr3p, and it could confer azole resistance in S. cerevisiae pdr1 pdr3 mutant by regulating PDR5. 16) We hypothesized that Fcr1p maybe involved in the development of C. albicans azole resistance as well as Pdr1p and Pdr3p in S. cerevisiae.…”

mentioning

confidence: 99%

Fcr1p Inhibits Development of Fluconazole Resistance in Candida albicans by Abolishing CDR1 Induction

Shen

Wang

et al. 2007

Biological & Pharmaceutical Bulletin

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Candida albicans (C. albicans), the major opportunistic fungal pathogen in humans, causes diseases varying from superficial mucosal complaints to life-threatening systemic disorders. [1][2][3] Coincident with the increased use of antifungal drugs, the incidences of drug resistance have also increased. [4][5][6] In recent years, the development of multidrug resistance in clinical isolates has challenged effective treatment of the infections. Specifically, the extensive and repetitive use of antifungal azole derivatives such as fluconazole (FLC) has allowed C. albicans to utilize many mechanisms of resistance in order to ensure its survival. 7,8) This situation highlights the need for elucidating the mechanism of drug resistance in C. albicans to develop new antifungal agents.Overexpression of efflux pumps, either ATP-binding cassette (ABC) or major facilitator superfamily (MFS) transporters, has been shown to be one of the major mechanisms of drug resistance in clinical isolates because if causes active extrusion of the drug out of the cell. 9-11) The Candida drug resistance 1 (CDR1) gene, which encodes an ABC efflux pump, is identified by complementation of the pleiotropic drug resistance 5 (pdr5) mutant, which is hypersensitive to cycloheximide, chloramphenicol, and azole drugs, in Saccharomyces cerevisiae (S. cerevisiae). 12) C. albicans cdr1 mutant resulted in increasing susceptibilities to azole drugs, 13) which is consistent with the observation that overexpression of CDR1 contributes to the drug resistance of clinical isolates of C. albicans. 6,14) Furthermore, the existence of trans-regulatory factors of CDR1 has also been suggested. 15) However, the molecular mechanism and the gene network regulating the expression of CDR1 and drug resistance are poorly understood.C. albicans Fcr1p (for fluconazole resistance 1 protein), a member of the family of zinc cluster proteins, is characterized by a highly conserved Zn(II) 2 Cys 6 zinc finger motif within the N-terminal DNA binding domain (DBD). 16) Well known Pdr1p and Pdr3p (pleiotropic drug resistance protein) zinc cluster proteins regulate the expression of several multidrug ABC transporter genes including PDR5, SNQ2, and YOR1, causing azole resistance in S. cerevisiae. 17-28) C. albicans Fcr1p displays significant sequence homology with S. cerevisiae Pdr1p and Pdr3p, and it could confer azole resistance in S. cerevisiae pdr1 pdr3 mutant by regulating PDR5. 16) We hypothesized that Fcr1p maybe involved in the development of C. albicans azole resistance as well as Pdr1p and Pdr3p in S. cerevisiae.In the present study we investigated the role of Fcr1p in the development of C. albicans azole resistance and possible mechanisms in vitro and in vivo. The results showed that Fcr1p inhibited development of fluconazole resistance in C. albicans through abolishing the induction of CDR1 expression by FLC, and in contrast FLC resistance development was accelerated resulting from the deletion of FCR1. (Table 1). MATERIALS AND METHODS Strains and MediaThe strains were cul...

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PDR3, a new yeast regulatory gene, is homologous toPDR1 and controls the multidrug resistance phenomenon

Cited by 203 publications

References 56 publications

ATPase and Multidrug Transport Activities of the Overexpressed Yeast ABC Protein Yor1p

ATPase and Multidrug Transport Activities of the Overexpressed Yeast ABC Protein Yor1p

Weak Organic Acids Trigger Conformational Changes of the Yeast Transcription Factor War1 in Vivo to Elicit Stress Adaptation

Fcr1p Inhibits Development of Fluconazole Resistance in Candida albicans by Abolishing CDR1 Induction

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