In vivo roles of Bm3R1 repressor in the barbiturate-mediated induction of the cytochrome P450 genes ( P450 BM-3 and P450 BM-1) of Bacillus megaterium

Abstract

We previously showed [Q. Liang, A.J. Fulco, J. Biol. Chem., 270 (1995) 18606–18614) that the binding of Bm3R1 repressor to Barbie box elements and operator sites in the 5′-flanking regions of the P450 BM-3 and P450 BM-1 ( CYP102 and CYP106) genes in Bacillus megaterium was a critical factor in their regulation at the level of transcription. We now describe experiments that delineate specific roles for Bm3R1 in the barbiturate-mediated induction of these genes. We directly demonstrate the interaction of Bm3R1 with Barbie box and operator sequences and show that high in vivo levels of Bm3R1 prevent putative positive factors from binding to Barbie box elements, strongly inhibit the expression of the P450 genes, prolong the lag phase of growth in Bacillus megaterium cultures and increase the sensitivity of the cells to the growth-inhibitory effects of barbiturates. Finally, our data suggest that there may be two forms of Bm3R1, either of which can interact with O III, the bicistronic operator sequence.