The activation of pectin lyase (Pnl) production in subsp. strain 71 occurs upon DNA damage via a unique regulatory circuit involving and In a similar Pnl-inducible system reconstituted in , the product was found to activate the expression of , the structural gene for pectin lyase. The kinetic data presented here also show that transcription of followed that of in subsp. , indicating a temporal order of gene expression. By deletion analysis we have localized the promoter/regulatory region within a 66 bp DNA segment upstream of the transcriptional start site. This region contains the -10 consensus sequence but not the sequences corresponding to the -35 region. For DNA-binding studies, was overexpressed in and a 14 kDa polypeptide was identified as the gene product. RdgB from crude extracts or a purified preparation caused an identical gel mobility shift of a 164 bp DNA segment containing the promoter/regulatory region. Utilizing DNase I protection assay the RdgB-binding site was localized between nucleotides -29 and -56, i.e. overlapping the position of the putative -35 box. The findings reported here, taken along with our previous observation that the product is required for expression, establishes that encodes a transcriptional factor which specifically interacts with the promoter/regulatory region.


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