A DNA microarray-based approach to elucidate the effects of the immunosuppressant SR31747A on gene expression in Saccharomyces cerevisiae.
SR31747A is an immunosuppressive agent that arrests cell proliferation in the yeast Saccharomyces cerevisiae. In this microorganism, SR31747A was shown to inhibit the ERG2 gene product, namely the delta8-delta7 sterol isomerase, involved in the ergosterol biosynthesis pathway. Although previous genetic experiments pointed to this enzyme as the target for SR31747A in yeast, the existence of other potential targets could not be ruled out. To enlighten this issue, we undertook a DNA microarray-based approach in which the expression profile of SR31747A-treated wild-type cells defining the "drug signature" was compared with the "mutant signature," the expression profile of the corresponding ERG2-deleted strain. We observed that treatment of ERG2-positive cells with SR31747A resulted in the modulation of mRNA levels of numerous genes. Among them, 121 werealso affected in untreated ERG2-disrupted cells compared with wild-type cells. By contrast, drug exposure did not induce any significant transcriptional change in the ERG2 null mutant. These results were consistent with SR31747A being an inhibitor of the sterol isomerase and demonstrated the absence of any additional SR31747A target. The detailed analysis of the observed 121 modulated genes provides new insights into the cellular response to ergosterol deprivation induced by SR31747A through inhibition of the ERG2 gene product.
Cinato
Elisa
E
Immunology-Oncology Department, Sanofi-Synthelabo, 371 rue du Professeur Joseph Blayac, Montpellier, France.
Péléraux
Annick
A
Silve
Sandra
S
Galiègue
Sylvaine
S
Dhers
Christiane
C
Picard
Claudine
C
Jbilo
Omar
O
Loison
Gérard
G
Casellas
Pierre
P
D015152:Blotting, Northern; D002455:Cell Division; D003510:Cyclohexanes / Q000494:pharmacology*; D004268:DNA-Binding Proteins / Q000378:metabolism; D015536:Down-Regulation; D000072237:ERG1 Potassium Channel; D004875:Ergosterol / Q000378:metabolism; D051638:Ether-A-Go-Go Potassium Channels; D008401:Gas Chromatography-Mass Spectrometry; D015966:Gene Expression Regulation, Fungal; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D008954:Models, Biological; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger; D012441:Saccharomyces cerevisiae / Q000187:drug effects*; D015534:Trans-Activators / Q000378:metabolism; D000071230:Transcriptional Regulator ERG; D015854:Up-Regulation
D003510:Cyclohexanes; D004268:DNA-Binding Proteins; C072312:ERG protein, human; D000072237:ERG1 Potassium Channel; D051638:Ether-A-Go-Go Potassium Channels; D007166:Immunosuppressive Agents; D012333:RNA, Messenger; D015534:Trans-Activators; D000071230:Transcriptional Regulator ERG; D012313:RNA; C088447:SR 31747; D004875:Ergosterol