Isolation and Analysis of the Acetate Regulatory Gene, facB, from Aspergillus nidulans

Abstract

The facB gene of Aspergillus nidulans is thought to be involved in acetate induction of enzymes required for acetate utilization and of the acetamidase encoded by the multiply regulated amdS gene. In addition, some evidence suggests that the facB gene has a structural as well as a regulatory role in acetate metabolism. The facB gene was cloned from a cosmid library by complementation of the facB 101 loss-of-function mutation. Transformants receiving multiple copies of facB displayed stronger growth on acetamide media, indicating increased amdS expression, while growth on acetate was inhibited in these multicopy transformants. A 3.1-kilobase acetate-inducible facB transcript was detected by Northern (RNA) blot analysis. Examination of message levels in wild-type and mutant strains indicated that the facB gene is subject to carbon catabolite repression. Previous work has indicated that the presence of multiple copies of the 5′ end of the amdS gene can result in titration of regulatory proteins. Additional copies of the facB gene were shown to specifically overcome the effect of facB product titration.