LexA, a transcription regulator binding in the promoter region of the bidirectional hydrogenase in the cyanobacterium Synechocystis sp. PCC 6803

The unicellular cyanobacterium Synechocystis PCC 6803 contains a single pentameric bidirectional hydrogenase encoded by hoxEFUYH. Transcriptional experiments demonstrated that the five hox genes are part of a single transcript together with three ORFs with unknown functions. The transcription start point was localized by 5′ RACE to 168bp upstream the hoxE ATG start codon. DNA affinity assays demonstrated a specific interaction between the hox regulatory promoter region and a protein which, using mass spectrometry, was identified to be LexA. Overexpressed His-tagged Synechocystis LexA and EMSA showed a specific binding to the promoter region of the hox operon. Increasing concentrations of the purified LexA resulted in two retarded LexA-DNA complexes, in agreement with the presence of two putative LexA binding sites upstream the determined TSP.