Relationship between chloroquine toxicity and iron acquisition in Saccharomyces cerevisiae.
Chloroquine is one of the most effective antimalarials, but resistance to it is becoming widespread. However, we do not fully understand either the drug's mode of action or the mechanism of resistance. In an effort to expand our understanding of the mechanism of action and resistance associated with chloroquine, we used Saccharomyces cerevisiae as a model eukaryotic system. To aid in the discovery of potential drug targets we applied the transcriptional profiling method to identify genes transcriptionally responsive to chloroquine treatment in S. cerevisiae. Among the genes that were differentially expressed with chloroquine treatment were a number of metal transporters involved in iron acquisition (SIT1, ARN2, ARN4, and SMF2). These genes exhibit similar expression patterns, and several are known to be regulated by AFT1, a DNA binding protein, which responds to iron levels in the cell. We investigated the role of chloroquine in iron metabolism by using a variety of approaches, including pharmacological, genetic, and biochemical techniques. For these experiments, we utilized yeast lacking the major iron uptake pathways (FET3 and FET4) and yeast deficient in SIT1, encoding the major up-regulated iron siderophore transporter. Our experiments show that yeast genetically or environmentally limited in iron availability has increased sensitivity to chloroquine in pharmacological assays and that the addition of iron rescues these cells from chloroquine killing. 55FeCl3 accumulation was inhibited in the presence of chloroquine, and kinetic analysis demonstrated that inhibition was competitive. These results are consistent with deprivation of iron as a mechanism of chloroquine killing in yeast.
Emerson
Lyndal R
LR
Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, Massachusetts 02115, USA.
Nau
Martin E
ME
Martin
Rodger K
RK
Kyle
Dennis E
DE
Vahey
Maryanne
M
Wirth
Dyann F
DF
D000962:Antimalarials / Q000633:toxicity*; D015152:Blotting, Northern; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002738:Chloroquine / Q000633:toxicity*; D019295:Computational Biology; D003470:Culture Media; D007501:Iron / Q000378:metabolism*; D007504:Iron Radioisotopes; D007700:Kinetics; D026901:Membrane Transport Proteins; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2; D012331:RNA, Fungal / Q000096:biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D017262:Siderophores / Q000235:genetics / Q000378:metabolism
D000962:Antimalarials; D002352:Carrier Proteins; D003470:Culture Media; D007504:Iron Radioisotopes; D026901:Membrane Transport Proteins; D012331:RNA, Fungal; D012333:RNA, Messenger; C118510:SIT1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D017262:Siderophores; D002738:Chloroquine; D007501:Iron; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2; C104343:SIT4 protein, S cerevisiae