A reporter gene assay for fungal sterol biosynthesis inhibitors.
Acetoacetyl-CoA thiolase (ACoAT) catalyses the condensation of two acetyl-CoA molecules, the first step in the sterol biosynthetic pathway. We constructed a yeast strain containing a fusion of the promoter of the Saccharomyces cerevisiae ACoAT gene to a reporter gene (Escherichia coli beta-galactosidase). Reporter gene activity in this strain can be induced by a variety of inhibitors of sterol biosynthesis. These results suggest that the ACoAT gene is feedback regulated at the transcriptional level by products of the sterol biosynthetic pathway. The reporter gene approach described here may be used to screen chemical collections for compounds which inhibit fungal sterol biosynthesis.
Dixon
G
G
Target Discovery and Infection Department, Zeneca Pharmaceuticals, Macclesfield, Cheshire, U.K.
Scanlon
D
D
Cooper
S
S
Broad
P
P
D000101:Acetyl-CoA C-Acetyltransferase / Q000235:genetics* / Q000378:metabolism; D001681:Biological Assay; D004926:Escherichia coli / Q000235:genetics; D017930:Genes, Reporter; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013261:Sterols / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D001616:beta-Galactosidase / Q000235:genetics
D013261:Sterols; D000101:Acetyl-CoA C-Acetyltransferase; D001616:beta-Galactosidase