Phosphatidylinositol binding of Saccharomyces cerevisiae Pdr16p represents an essential feature of this lipid transfer protein to provide protection against azole antifungals

Roman Holič; Zuzana Simová; Tim Ashlin; Vladimír Pevala; Katarína Poloncová; Dana Tahotná; Eva Kutejová; Shamshad Cockcroft; Peter Griač

doi:10.1016/j.bbalip.2014.07.014

Phosphatidylinositol binding of Saccharomyces cerevisiae Pdr16p represents an essential feature of this lipid transfer protein to provide protection against azole antifungals

Biochim Biophys Acta. 2014 Oct;1842(10):1483-90. doi: 10.1016/j.bbalip.2014.07.014. Epub 2014 Jul 25.

Authors

Roman Holič¹, Zuzana Simová¹, Tim Ashlin², Vladimír Pevala³, Katarína Poloncová¹, Dana Tahotná¹, Eva Kutejová³, Shamshad Cockcroft², Peter Griač⁴

Affiliations

¹ Department of Membrane Biochemistry, Institute of Animal Biochemistry and Genetics, Slovak Academy of Sciences, 900 28 Ivanka pri Dunaji, Slovakia.
² Department of Neuroscience, Physiology and Pharmacology, Division of Biosciences, University College London, London WC1E 6JJ, United Kingdom.
³ Department of Biochemistry and Structural Biology, Institute of Molecular Biology, Slovak Academy of Sciences, 845 51 Bratislava, Slovakia.
⁴ Department of Membrane Biochemistry, Institute of Animal Biochemistry and Genetics, Slovak Academy of Sciences, 900 28 Ivanka pri Dunaji, Slovakia. Electronic address: Peter.Griac@savba.sk.

Abstract

Pdr16p is considered a factor of clinical azole resistance in fungal pathogens. The most distinct phenotype of yeast cells lacking Pdr16p is their increased susceptibility to azole and morpholine antifungals. Pdr16p (also known as Sfh3p) of Saccharomyces cerevisiae belongs to the Sec14 family of phosphatidylinositol transfer proteins. It facilitates transfer of phosphatidylinositol (PI) between membrane compartments in in vitro systems. We generated Pdr16p(E235A, K267A) mutant defective in PI binding. This PI binding deficient mutant is not able to fulfill the role of Pdr16p in protection against azole and morpholine antifungals, providing evidence that PI binding is critical for Pdr16 function in modulation of sterol metabolism in response to these two types of antifungal drugs. A novel feature of Pdr16p, and especially of Pdr16p(E235A, K267A) mutant, to bind sterol molecules, is observed.

Keywords: Azole resistance; Lipid binding; SFH3; Saccharomyces cerevisiae; Sterol metabolism.