The antidepressant sertraline targets intracellular vesiculogenic membranes in yeast.
Numerous studies have shown that the clinical antidepressant sertraline (Zoloft) is biologically active in model systems, including fungi, which do not express its putative protein target, the serotonin/5-HT transporter, thus demonstrating the existence of one or more secondary targets. Here we show that in the absence of its putative protein target, sertraline targets phospholipid membranes that comprise the acidic organelles of the intracellular vesicle transport system by a mechanism consistent with the bilayer couple hypothesis. On the basis of a combination of drug-resistance selection and chemical-genomic screening, we hypothesize that loss of vacuolar ATPase activity reduces uptake of sertraline into cells, whereas dysregulation of clathrin function reduces the affinity of membranes for sertraline. Remarkably, sublethal doses of sertraline stimulate growth of mutants with impaired clathrin function. Ultrastructural studies of sertraline-treated cells revealed a phenotype that resembles phospholipidosis induced by cationic amphiphilic drugs in mammalian cells. Using reconstituted enzyme assays, we also demonstrated that sertraline inhibits phospholipase A(1) and phospholipase D, exhibits mixed effects on phospholipase C, and activates phospholipase A(2). Overall, our study identifies two evolutionarily conserved membrane--active processes-vacuolar acidification and clathrin-coat formation--as modulators of sertraline's action at membranes.
Rainey
Meredith M
MM
Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey 08544, USA.
Korostyshevsky
Daniel
D
Lee
Sean
S
Perlstein
Ethan O
EO
D000928:Antidepressive Agents / Q000494:pharmacology; D002966:Clathrin / Q000235:genetics / Q000378:metabolism; D022162:Cytoplasmic Vesicles / Q000187:drug effects* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004351:Drug Resistance / Q000235:genetics; D007425:Intracellular Membranes / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D008563:Membrane Lipids / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D009154:Mutation; D010740:Phospholipases / Q000037:antagonists & inhibitors / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D020280:Sertraline / Q000494:pharmacology*; D025262:Vacuolar Proton-Translocating ATPases / Q000235:genetics / Q000378:metabolism; D014617:Vacuoles / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure
D000928:Antidepressive Agents; D002966:Clathrin; D008563:Membrane Lipids; D010743:Phospholipids; D029701:Saccharomyces cerevisiae Proteins; D010740:Phospholipases; D025262:Vacuolar Proton-Translocating ATPases; D020280:Sertraline
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