Developmental regulation of laccase levels in Aspergillus nidulans

Abstract

Asexual spores (conidia) of Aspergillus nidulans contain a dark green pigment which is not present in other cell types. Synthesis of this pigment is catalyzed, in part, by a developmentally controlled p-diphenol oxidase, or laccase, encoded at the gamma A genetic locus (A. J. Clutterbuck, J. Gen. Microbiol. 70:423-435, 1972). We have investigated the mechanisms regulating expression of the gamma A gene of A. nidulans. Vegetative hyphae grown in submerged culture lacked detectable laccase enzyme activity and neither contained nor synthesized immunoprecipitable laccase protein. When such cultures were induced to conidiate by harvesting the cells onto filter papers and aerating them, laccase levels began to increase after 10 to 16 h, reached a peak at 20 to 36 h, and then declined slowly. Immunological assays showed that increases in laccase enzyme activity were (i) proceded by a transient rise in the relative rate of laccase protein synthesis and (ii) closely paralleled by increases in the amount of laccase protein. Addition of cycloheximide to cultures at any time after inducing conidiation inhibited further accumulation of laccase enzyme activity. These data are most consistent with increases in laccase levels being due to regulated, de novo synthesis of laccase protein. Addition of inhibitors of ribonucleic acid synthesis to conidiating cultures also inhibited further accumulation of laccase, suggesting that laccase expression is regulated by alterations in the transcriptional activity of the gamma A locus.