Novel Insights into the Regulation of LexA in the Cyanobacterium Synechocystis sp. Strain PCC 6803

Abstract

ABSTRACT The transcription factor LexA in the cyanobacterium Synechocystis sp. strain PCC 6803 has been shown to regulate genes that are not directly involved in DNA repair but instead in several different metabolic pathways. However, the signal transduction pathways remain largely uncharacterized. The present work gives novel insights into the regulation of LexA in this unicellular cyanobacterium. A combination of Northern and Western blotting, using specific antibodies against the cyanobacterial LexA, was employed to show that this transcription regulator is under posttranscriptional control, in addition to the classical and already-described transcriptional regulation. Moreover, detailed two-dimensional (2D) electrophoresis analyses of the protein revealed that LexA undergoes posttranslational modifications. Finally, a fully segregated LexA::GFP (green fluorescent protein) fusion-modified strain was produced to image LexA's spatial distribution in live cells. The fusion protein retains DNA binding capabilities, and the GFP fluorescence indicates that LexA is localized in the innermost region of the cytoplasm, decorating the DNA in an evenly distributed pattern. The implications of these findings for the overall role of LexA in Synechocystis sp. strain PCC 6803 are further discussed.