A high affinity glutathione transporter has been identified, cloned, and characterized from the yeast Saccharomyces cerevisiae. This transporter, Hgt1p, represents the first high affinity glutathione transporter to be described from any system so far. The strategy for the identification involved investigating candidate glutathione transporters from the yeast genome sequence project followed by genetic and physiological investigations. This approach revealed HGT1 (open reading frame YJL212c) as encoding a high affinity glutathione transporter. Yeast strains deleted in HGT1 did not show any detectable plasma membrane glutathione transport, and hgt1⌬ disruptants were non-viable in a glutathione biosynthetic mutant (gsh1⌬) background. The glutathione repressible transport activity observed in wild type cells was also absent in the hgt1⌬ strains. The transporter was cloned and kinetic studies indicated that Hgt1p had a high affinity for glutathione (K m ‫؍‬ 54 M)) and was not sensitive to competition by amino acids, dipeptides, or other tripeptides. Significant inhibition was observed, however, with oxidized glutathione and glutathione conjugates. The transporter reveals a novel class of transporters that has homologues in other yeasts and plants but with no apparent homologues in either Escherichia coli or in higher eukaryotes other than plants.

The enzyme Psd2 catalyzes endosomal synthesis of the phospholipid PE. While this pool of PE represents a minority of total cellular PE, function of Psd2 is required for normal activity of the vacuolar ABC transporter Ycf1. Psd2 controls vacuolar PE levels by acting at the level of the endosome.

Multidrug resistance in the yeast

Med12 is a transcriptional Mediator subunit most typically associated with negative control of gene expression. Here Med12 is demonstrated to serve as a positive regulator required for activation of multidrug resistance gene expression in yeast cells lacking their mitochondrial genome.

Summary Pathogenic fungi present a special problem in the clinic as the range of drugs that can be used to treat these types of infections is limited. This situation is further complicated by the presence of robust inducible gene networks encoding different proteins that confer tolerance to many available antifungal drugs. The transcriptional control of these multidrug resistance systems in several key fungi will be discussed. Experiments in the non-pathogenic Saccharomyces cerevisiae have provided much of our current understanding of the molecular framework on which fungal multidrug resistance is built. More recent studies on the important pathogenic Candida species, Candida albicans and Candida glabrata, have provided new insights into the organization of the multidrug resistance systems in these organisms. We will compare the circuitry of multidrug resistance networks in these three organisms and suggest that, in addition to the well-accepted drug efflux activities, the regulation of membrane composition by multidrug resistance proteins provides an important contribution to the resistant phenotypes observed.