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LexA Binds to Transcription Regulatory Site of Cell Division Gene ftsZ in Toxic Cyanobacterium Microcystis aeruginosa

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Abstract

Previously, we showed that DNA replication and cell division in toxic cyanobacterium Microcystis aeruginosa are coordinated by transcriptional regulation of cell division gene ftsZ and that an unknown protein specifically bound upstream of ftsZ (BpFz; DNA-binding protein to an upstream site of ftsZ) during successful DNA replication and cell division. Here, we purified BpFz from M. aeruginosa strain NIES-298 using DNA-affinity chromatography and gel-slicing combined with gel electrophoresis mobility shift assay (EMSA). The N-terminal amino acid sequence of BpFz was identified as TNLESLTQ, which was identical to that of transcription repressor LexA from NIES-843. EMSA analysis using mutant probes showed that the sequence GTACTAN3GTGTTC was important in LexA binding. Comparison of the upstream regions of lexA in the genomes of closely related cyanobacteria suggested that the sequence TASTRNNNNTGTWC could be a putative LexA recognition sequence (LexA box). Searches for TASTRNNNNTGTWC as a transcriptional regulatory site (TRS) in the genome of M. aeruginosa NIES-843 showed that it was present in genes involved in cell division, photosynthesis, and extracellular polysaccharide biosynthesis. Considering that BpFz binds to the TRS of ftsZ during normal cell division, LexA may function as a transcriptional activator of genes related to cell reproduction in M. aeruginosa, including ftsZ. This may be an example of informality in the control of bacterial cell division.

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Funding

This study was partly supported by a Grant-in-Aid for Science Research B (No. 20310045, No. 23310056 and No. 17H03850) from The Japan Society for the Promotion of Science (JSPS).

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Correspondence to Takashi Yoshida.

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Table S1

Putative LexA box and LexA-regulated genes in Microcystis. aTranscription units were predicted using FGENESB (http://www.softberry.com/berry) (Solovyev and Salamov 2011). bGene number from CyanoBase (http://genome.microbedb.jp/cyanobase). cDescription of gene function as per the CyanoBase. dThe LexA box is indicated in uppercase letters. eThe position of the LexA box is indicated upstream (−) or downstream (+) of the start codon (+1). fThe direction of the LexA box is indicated in relation to the gene direction (forward, D; opposite, R). (XLSX 13 kb).

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Honda, T., Morimoto, D., Sako, Y. et al. LexA Binds to Transcription Regulatory Site of Cell Division Gene ftsZ in Toxic Cyanobacterium Microcystis aeruginosa. Mar Biotechnol 20, 549–556 (2018). https://doi.org/10.1007/s10126-018-9826-4

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