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A nuclear receptor-like pathway regulating multidrug resistance in fungi

Abstract

Multidrug resistance (MDR) is a serious complication during treatment of opportunistic fungal infections that frequently afflict immunocompromised individuals, such as transplant recipients and cancer patients undergoing cytotoxic chemotherapy. Improved knowledge of the molecular pathways controlling MDR in pathogenic fungi should facilitate the development of novel therapies to combat these intransigent infections. MDR is often caused by upregulation of drug efflux pumps by members of the fungal zinc-cluster transcription-factor family (for example Pdr1p orthologues). However, the molecular mechanisms are poorly understood. Here we show that Pdr1p family members in Saccharomyces cerevisiae and the human pathogen Candida glabrata directly bind to structurally diverse drugs and xenobiotics, resulting in stimulated expression of drug efflux pumps and induction of MDR. Notably, this is mechanistically similar to regulation of MDR in vertebrates by the PXR nuclear receptor, revealing an unexpected functional analogy of fungal and metazoan regulators of MDR. We have also uncovered a critical and specific role of the Gal11p/MED15 subunit of the Mediator co-activator and its activator-targeted KIX domain in antifungal/xenobiotic-dependent regulation of MDR. This detailed mechanistic understanding of a fungal nuclear receptor-like gene regulatory pathway provides novel therapeutic targets for the treatment of multidrug-resistant fungal infections.

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Figure 1: Pdr1p is a xenobiotic receptor.
Figure 2: Requirement for the Gal11p Mediator subunit and its KIX domain in Pdr1p/Pdr3p-dependent transcription of target genes and MDR.
Figure 3: Structure of the Gal11p KIX domain and structural/mutational analysis of the Pdr1pAD-Gal11p KIX interface.
Figure 4: Dissection of the molecular mechanism of drug resistance in C. glabrata.

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Protein Data Bank

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The NMR structure is deposited in the Protein Data Bank under accession number 2k0n.

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Acknowledgements

We thank C. Wivagg and M. Franzmann for their help with the NMR assignment and fluorescence studies; and N. Dyson, S. Buratowski, A. Walker and H. Najafi-Shoushtari for comments on the manuscript. We were unable to cite many original papers owing to space constraints. This work was supported by grants from the National Institutes of Health: GM071449 (A.M.N.), CA127990 (G.W./A.M.N.), A1046223 (B.P.C.), GM49825 (W.S.M.-R.) and GM30186 (K.S.). The NMR facility used for this research was supported by grants GM47467 and EB2026. J.K.T. was supported by a fellowship from the MGH Fund for Medical Discovery.

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Correspondence to Anders M. Näär.

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Supplementary Information

The file contains Supplementary Figures 1-11 with Legends and Supplementary Tables 1-3 The Supplementary Figures describe data in support of the main findings presented in the print version of the paper. The Supplementary Table 1 shows the fungal strains used in the paper, the Supplementary Table 2 lists the primers used for quantitative real-time RT-PCR, and the Supplementary Table 3 outlines the structural NMR data and parameters. (PDF 2504 kb)

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Thakur, J., Arthanari, H., Yang, F. et al. A nuclear receptor-like pathway regulating multidrug resistance in fungi. Nature 452, 604–609 (2008). https://doi.org/10.1038/nature06836

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