Graduate school of Agriculture, Kyoto University, Kitashirakawa-Oiwake, Sakyo-ku, Kyoto, 606-8502, Japan.
Mar Biotechnol (NY). 2018 Aug;20(4):549-556. doi: 10.1007/s10126-018-9826-4. Epub 2018 May 17.
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 GTACTANGTGTTC 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.
先前,我们表明,有毒蓝藻微囊藻中 DNA 复制和细胞分裂是通过细胞分裂基因 ftsZ 的转录调控协调的,并且在成功的 DNA 复制和细胞分裂期间,一种未知的蛋白质(BpFz;ftsZ 上游位点的 DNA 结合蛋白)特异性地结合在 ftsZ 的上游。在这里,我们使用 DNA 亲和层析和凝胶切片结合凝胶电泳迁移率变动分析(EMSA)从 M. aeruginosa 株 NIES-298 中纯化 BpFz。BpFz 的 N 末端氨基酸序列被鉴定为 TNLESLTQ,与 NIES-843 中的转录阻遏物 LexA 相同。使用突变探针的 EMSA 分析表明,序列 GTACTANGTGTTC 对 LexA 结合很重要。比较密切相关的蓝藻基因组中 lexA 的上游区域表明,序列 TASTRNNNNTGTWC 可能是一个推定的 LexA 识别序列(LexA 盒)。在 M. aeruginosa NIES-843 的基因组中搜索 TASTRNNNNTGTWC 作为转录调控位点(TRS)的结果表明,它存在于参与细胞分裂、光合作用和细胞外多糖生物合成的基因中。考虑到 BpFz 在正常细胞分裂期间结合 ftsZ 的 TRS,LexA 可能作为 M. aeruginosa 中与细胞繁殖相关的基因(包括 ftsZ)的转录激活物发挥作用。这可能是细菌细胞分裂控制不规范的一个例子。