PDR5, a novel yeast multidrug resistance conferring transporter controlled by the transcription regulator PDR1.

Balzi, Elisabetta; Wang, M; Leterme, S.; Dyck, L Van; Goffeau, André

doi:10.1016/s0021-9258(17)42155-7

Cited by 400 publications

(39 citation statements)

References 74 publications

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“…An in vivo study focusing on treatment of melanoma brain metastases using dabrafenib revealed that P-glycoprotein and breast cancer resistance protein efficiently pump dabrafenib to the extracellular matrix [64]. Since P-glycoprotein belongs to an ATP-binding cassette (ABC) sub-family [65], we speculate that yeast ABC family proteins, such as PDR-encoded multidrug resistance transporter [66], may pump dabrafenib out of yeast cells, limiting its intracellular drug concentration in yeast. We are not aware of studies investigating whether membrane transporters can pump GSK'872 out of cells, so we cannot speculate on the likelihood that efflux of GSK'872 from yeast accounts for its lack of activity in this model.…”

Section: Discussionmentioning

confidence: 93%

Reconstitution of Human Necrosome Interactions in Saccharomyces cerevisiae

Ward

Hawkins

2021

Biomolecules

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The necrosome is a large-molecular-weight complex in which the terminal effector of the necroptotic pathway, Mixed Lineage Kinase Domain-Like protein (MLKL), is activated to induce necroptotic cell death. The precise mechanism of MLKL activation by the upstream kinase, Receptor Interacting Serine/Threonine Protein Kinase 3 (RIPK3) and the role of Receptor Interacting Serine/Threonine Protein Kinase 1 (RIPK1) in mediating MLKL activation remain incompletely understood. Here, we reconstituted human necrosome interactions in yeast by inducible expression of these necrosome effectors. Functional interactions were reflected by the detection of phosphorylated MLKL, plasma membrane permeabilization, and reduced proliferative potential. Following overexpression of human necrosome effectors in yeast, MLKL aggregated in the periphery of the cell, permeabilized the plasma membrane and compromised clonogenic potential. RIPK1 had little impact on RIPK3/MLKL-mediated yeast lethality; however, it exacerbated the toxicity provoked by co-expression of MLKL with a RIPK3 variant bearing a mutated RHIM-domain. Small molecule necroptotic inhibitors necrostatin-1 and TC13172, and viral inhibitors M45 (residues 1–90) and BAV_Rmil, abated the yeast toxicity triggered by the reconstituted necrosome. This yeast model provides a convenient tool to study necrosome protein interactions and to screen for and characterize potential necroptotic inhibitors.

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Section: Discussionmentioning

confidence: 93%

Reconstitution of Human Necrosome Interactions in Saccharomyces cerevisiae

Ward

Hawkins

2021

Biomolecules

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“…We next examined Pdr5, another integral PM protein whose transcript is a Puf1 and Puf2 target [ 15 , 17 ]. Pdr5 is a large (1511 residues) multi-drug transporter in the ABC-family with 12 transmembrane segments [ 69 ]. To monitor Pdr5, we utilized a rabbit polyclonal antibody against native Pdr5, generously provided by Prof. Karl Kuchler (Medical University of Vienna, Austria).…”

Section: Resultsmentioning

confidence: 99%

Phosphorylation of mRNA-Binding Proteins Puf1 and Puf2 by TORC2-Activated Protein Kinase Ypk1 Alleviates Their Repressive Effects

et al. 2021

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Members of the Puf family of RNA-binding proteins typically associate via their Pumilio homology domain with specific short motifs in the 3’-UTR of an mRNA and thereby influence the stability, localization and/or efficiency of translation of the bound transcript. In our prior unbiased proteome-wide screen for targets of the TORC2-stimulated protein kinase Ypk1, we identified the paralogs Puf1/Jsn1 and Puf2 as high-confidence substrates. Earlier work by others had demonstrated that Puf1 and Puf2 exhibit a marked preference for interaction with mRNAs encoding plasma membrane-associated proteins, consistent with our previous studies documenting that a primary physiological role of TORC2-Ypk1 signaling is maintenance of plasma membrane homeostasis. Here, we show, first, that both Puf1 and Puf2 are authentic Ypk1 substrates both in vitro and in vivo. Fluorescently tagged Puf1 localizes constitutively in cortical puncta closely apposed to the plasma membrane, whereas Puf2 does so in the absence of its Ypk1 phosphorylation, but is dispersed in the cytosol when phosphorylated. We further demonstrate that Ypk1-mediated phosphorylation of Puf1 and Puf2 upregulates production of the protein products of the transcripts to which they bind, with a concomitant increase in the level of the cognate mRNAs. Thus, Ypk1 phosphorylation relieves Puf1- and Puf2-mediated post-transcriptional repression mainly by counteracting their negative effect on transcript stability. Using a heterologous protein-RNA tethering and fluorescent protein reporter assay, the consequence of Ypk1 phosphorylation in vivo was recapitulated for full-length Puf1 and even for N-terminal fragments (residues 1-340 and 143-295) corresponding to the region upstream of its dimerization domain (an RNA-recognition motif fold) encompassing its two Ypk1 phosphorylation sites (both also conserved in Puf2). This latter result suggests that alleviation of Puf1-imposed transcript destabilization does not obligatorily require dissociation of Ypk1-phosphorylated Puf1 from a transcript. Our findings add new insight about how the TORC2-Ypk1 signaling axis regulates the content of plasma membrane-associated proteins to promote maintenance of the integrity of the cell envelope.

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“…The zinc cluster transcription factors Pdr1 (Balzi et al, 1994) and Tac1 (Coste et al, 2004;Liu et al, 2018) are responsible for the upregulation of the prototype fungal PDR transporter genes S. cerevisiae PDR5 and C. albicans CDR1, respectively. Zinc cluster transcription factors are only found in fungi.…”

Section: Discussionmentioning

confidence: 99%

PDR Transporter ABC1 Is Involved in the Innate Azole Resistance of the Human Fungal Pathogen Fusarium keratoplasticum

et al. 2021

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Fusarium keratoplasticum is arguably the most common Fusarium solani species complex (FSSC) species associated with human infections. Invasive fusariosis is a life-threatening fungal infection that is difficult to treat with conventional azole antifungals. Azole drug resistance is often caused by the increased expression of pleiotropic drug resistance (PDR) ATP-binding cassette (ABC) transporters of the ABCG sub-family. Most investigations of Fusarium ABC transporters associated with azole antifungal drug resistance are limited to plant pathogens. Through the manual curation of the entire ABCG protein family of four FSSC species including the fully annotated genome of the plant pathogen Nectria haematococca we identified PDR transporters ABC1 and ABC2 as the efflux pump candidates most likely to be associated with the innate azole resistance phenotype of Fusarium keratoplasticum. An initial investigation of the transcriptional response of logarithmic phase F. keratoplasticum cells to 16 mg/L voriconazole confirmed strong upregulation (372-fold) of ABC1 while ABC2 mRNA levels were unaffected by voriconazole exposure over a 4 h time-period. Overexpression of F. keratoplasticum ABC1 and ABC2 in the genetically modified Saccharomyces cerevisiae host ADΔΔ caused up to ∼1,024-fold increased resistance to a number of xenobiotics, including azole antifungals. Although ABC1 and ABC2 were only moderately (20% and 10%, respectively) expressed compared to the Candida albicans multidrug efflux pump CDR1, overexpression of F. keratoplasticum ABC1 caused even higher resistance levels to certain xenobiotics (e.g., rhodamine 6G and nigericin) than CDR1. Our investigations suggest an important role for ABC1 orthologues in the innate azole resistance phenotype of FSSC species.

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PDR5, a novel yeast multidrug resistance conferring transporter controlled by the transcription regulator PDR1.

Cited by 400 publications

References 74 publications

Reconstitution of Human Necrosome Interactions in Saccharomyces cerevisiae

Reconstitution of Human Necrosome Interactions in Saccharomyces cerevisiae

Phosphorylation of mRNA-Binding Proteins Puf1 and Puf2 by TORC2-Activated Protein Kinase Ypk1 Alleviates Their Repressive Effects

PDR Transporter ABC1 Is Involved in the Innate Azole Resistance of the Human Fungal Pathogen Fusarium keratoplasticum

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