Genomewide expression profiling of cryptolepine-induced toxicity in Saccharomyces cerevisiae.
We have used the budding yeast Saccharomyces cerevisiae to identify genes that may confer sensitivity in vivo to the antimalarial and cytotoxic agent cryptolepine. Five S. cerevisiae strains, with different genetic backgrounds in cell permeability and DNA damage repair mechanisms, were exposed to several concentrations of cryptolepine. Cryptolepine showed a relatively mild toxicity for wild-type strains, which was augmented by either increasing cell permeability (Deltaerg6 or ISE2 strains) or disrupting DNA damage repair (Deltarad52 strains). These results are compatible with the ability of cryptolepine to intercalate into DNA and thus promote DNA lesions. The effects of low concentrations of cryptolepine (20% and 40% inhibitory concentrations [IC(20) and IC(40)]) were analyzed by comparing the gene expression profiles of treated and untreated Deltaerg6 yeast cells. Significant changes in expression levels were observed for 349 genes (117 upregulated and 232 downregulated). General stress-related genes constituted the only recognizable functional cluster whose expression was increased upon cryptolepine treatment, making up about 20% of upregulated genes. In contrast, analysis of the characteristics of downregulated genes revealed a specific effect of cryptolepine on genes related to iron transport or acid phosphatases, as well as a significant proportion of genes related to cell wall components. The effects of cryptolepine on the transcription of iron transport-related genes were consistent with a loss of function of the iron sensor Aft1p, indicating a possible disruption of iron metabolism in S. cerevisiae. Since the interference of cryptolepine with iron metabolism is considered one of its putative antimalarial targets, this finding supports the utility of S. cerevisiae in drug-developing schemes.
Rojas
Marta
M
Instituto de Biología Molecular de Barcelona, CSIC, Parc Científic de Barcelona, Barcelona, Spain.
Wright
Colin W
CW
Piña
Benjamin
B
Portugal
José
J
D000962:Antimalarials / Q000633:toxicity; D001483:Base Sequence; D002463:Cell Membrane Permeability / Q000235:genetics; D004249:DNA Damage; D017931:DNA Primers / Q000235:genetics; D004260:DNA Repair / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D025141:Drug Resistance, Fungal / Q000235:genetics; D020869:Gene Expression Profiling; D016681:Genome, Fungal; D026121:Indole Alkaloids / Q000633:toxicity*; D007364:Intercalating Agents / Q000633:toxicity; D007501:Iron / Q000378:metabolism; D008780:Methyltransferases / Q000235:genetics; D011804:Quinolines / Q000633:toxicity*; D051721:Rad52 DNA Repair and Recombination Protein / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D013312:Stress, Physiological / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
C090297:AFT1 protein, S cerevisiae; D000962:Antimalarials; D017931:DNA Primers; D004271:DNA, Fungal; D026121:Indole Alkaloids; D007364:Intercalating Agents; D011804:Quinolines; C497197:RAD52 protein, S cerevisiae; D051721:Rad52 DNA Repair and Recombination Protein; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C024015:cryptolepine; D007501:Iron; D008780:Methyltransferases; C022670:delta 24-sterol methyltransferase