Deletion Mutagenesis of the 5′ psbA2 Region in Synechocystis 6803: Identification of a Putative cis Element Involved in Photoregulation

Abstract

Three Synechocystis 6803 mutants, with deletions in the upstream region of the light-regulated psbA2 gene, were constructed. All three mutants grew photoautotrophically but they were affected in psbA2 expression. In one of the mutants, ΔU80, psbA2 transcription levels were not suppressed under low light conditions and production of psbA2 transcripts occurred also in darkness, in contrast to the situation in the control strain. Comparative sequence analyses of the mutants identified a hexanucleotide repeat as possible cis element in psbA2 light regulation. Electrophoresis mobility shift assays showed that protein binding to the hexanucleotide repeat decreased with light intensity, consistent with a hypothesis that the repeat harbors a binding site for a repressor protein. Repeats similar to that in psbA2 were found also in the upstream region of other light-regulated genes in Synechocystsis 6803. We refer to these hexanucleotide repeats as High Light Regulatory 1 (HLR1) sequences.