Sulphur and nitrogen regulation of the protease-encoding ACP1 gene in the fungus Botrytis cinerea: correlation with a phospholipase D activity.

Sulphur and nitrogen catabolic repressions are regulations that have long been recognized in fungi, but whose molecular bases remain largely elusive. This paper shows that catabolic repression of a protease-encoding gene correlates with the modulation of a phosphatidylethanolamine (PE)-specific phospholipase D (PLD) activity in the pathogenic fungus Botrytis cinerea. Our results first demonstrate that the ACP1 gene is subject to sulphur catabolic repression, with sulphate and cysteine inhibiting its expression. Sulphate and cysteine also cause a decrease of the total cellular PLD activity and, reciprocally, the two PLD inhibitors AEBSF [4-(2-aminoethyl)benzenesulphonyl fluoride] and curcumin negatively affect ACP1 expression in vivo. Cysteine moreover inhibits the PE-specific PLD activity in cell extracts. ACP1 is regulated by nitrogen, but here we show that this regulation does not rely on the proximal AREA binding site in its promoter, and that glutamine does not play a particular role in the process. A decrease in the total cellular PLD activity is also observed when the cells are fed ammonia, but this effect is smaller than that produced by sulphur. RNA-interference experiments finally suggest that the enzyme responsible for the PE-specific PLD activity is encoded by a gene that does not belong to the known HKD gene family of PLDs.