Specific interactions of steroids, arylhydrocarbons and flavonoids with progesterone receptors from the cytosol of the fungus Rhizopus nigricans

Abstract

Rhizopus nigricans ( R. nigricans) transforms fungitoxic progesterone into the less toxic 11α-hydroxyprogesterone which is then able to exit the mycelia into the surrounding water. Hydroxylation of progesterone is an inducible process in which cytosolic progesterone receptors could be involved. In the present study, we characterised receptors with respect to ligand specificity and to their involvement in progesterone induction of hydroxylase. EC 50 values of different ligands (steroids, xenobiotic arylhydrocarbons and natural flavonoids) were determined by competition studies using 40 nM ( 3H)progesterone. C21 and C19 3-oxo-4-ene steroids were good competitors (EC 50 of progesterone 2.3 ± 0.1 × 10 −7 M, EC 50 of androsten-3,17-dione 24 ± 2 × 10 −7 M). The presence of hydroxyl groups in steroids significantly decreased the affinity for receptors. The arylhydrocarbons α-naphthoflavone and ketoconazole exhibited EC 50 values of 0.3 ± 0.01 × 10 −7 M and 27 ± 5 × 10 −7 M, respectively, whereas β-naphthoflavone and benzo(a)pyrene were not able to displace labelled progesterone completely. The competition curves obtained by natural flavonoids also did not reach the bottom level of non-labelled progesterone, indicating the interaction at some allosteric binding site(s) of progesterone receptors. All ligands were examined for their involvement in progesterone-hydroxylase induction. Steroid agonists induced the enzyme in a dose-dependent manner in accordance with their affinity for receptors, whereas arylhydrocarbons and natural flavonoids did not induce the enzyme. The agonistic action of steroids, together with the antagonistic action of α-naphthoflavone, strongly suggests the involvement of progesterone receptors in progesterone signalling resulting in the induction of progesterone-hydroxylase.