Hgt1p, a high affinity glutathione transporter from the yeast Saccharomyces cerevisiae

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 hgt1Δ disruptants were non-viable in a glutathione biosynthetic mutant (gsh1Δ) background. The glutathione repressible transport activity observed in wild type cells was also absent in the hgt1Δ strains. The transporter was cloned and kinetic studies indicated that Hgt1p had a high affinity for glutathione (Km = 54 μM)) 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

Evidence for the Bifunctional Nature of Mitochondrial Phosphatidylserine Decarboxylase: Role in Pdr3-Dependent Retrograde Regulation of PDR5 Expression▿

Multidrug resistance in the yeast Saccharomyces cerevisiae is sensitive to the mitochondrial genome status of cells. Cells that lose their organellar genome ([rho0] cells) dramatically induce transcription of multiple or pleiotropic drug resistance genes via increased expression of a zinc cluster-containing transcription factor designated Pdr3. A major Pdr3 target gene is the ATP-binding cassette transporter-encoding gene PDR5. Pdr5 has been demonstrated to act as a phospholipid floppase catalyzing the net outward movement of phosphatidylethanolamine (PE). Since the mitochondrially localized Psd1 enzyme provides a major route of PE biosynthesis, we evaluated the potential linkage between Psd1 function and PDR5 regulation. Overproduction of Psd1 in wild-type ([rho+]) cells was found to induce PDR5 transcription and drug resistance in a Pdr3-dependent manner. Loss of the PSD1 gene from [rho0] cells prevented the normal activation of PDR5 expression. Surprisingly, expression of a catalytically inactive form of Psd1 still supported PDR5 transcriptional activation, suggesting that PE levels were not the signal triggering PDR5 induction. Expression of green fluorescent protein fusions mapped the region required to induce PDR5 expression to the noncatalytic amino-terminal portion of Psd1. Psd1 is a novel bifunctional protein required both for PE biosynthesis and regulation of multidrug resistance

A lysosome targetable fluorescent probe for endogenous imaging of hydrogen peroxide in living cells

A lysosome targetable naphthalimide based fluorescent probe (LyNC) has been designed and synthesized which detects hydrogen peroxide (H<SUB>2</SUB>O<SUB>2</SUB>) with high selectivity and sensitivity in brain tissues and in living nematodes among various ROS/RNS tested. Further, the probe LyNC was successfully employed in exogenous and endogenous imaging of H<SUB>2</SUB>O<SUB>2</SUB> in living cell lines

Compartment-specific Synthesis of Phosphatidylethanolamine Is Required for Normal Heavy Metal Resistance

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