Kizawa Ayumi, Osanai Takashi
Department of Agricultural Chemistry, School of Agriculture, Meiji University.
J Gen Appl Microbiol. 2020 Jun 17;66(2):121-128. doi: 10.2323/jgam.2020.01.004. Epub 2020 Mar 13.
In photosynthetic microorganisms, cell cycle progression depends on day and night cycles; however, how cell division is regulated in response to these environmental changes is poorly understood. RpaA has been implicated in the signal output from both circadian clocks and light/dark conditions in the unicellular spherical-celled cyanobacterium Synechocystis sp. PCC 6803. In the present study, we investigated the involvement of a two-component response regulator RpaA in cell division regulation. Firstly, we examined the effects of rpaA overexpression on cell morphology and the expression levels of cell division genes. We observed an increase in the volume of non-dividing cells and a high proportion of dividing cells in rpaA-overexpressing strains by light microscopy. The expression levels of selected cell division-related genes were higher in the rpaA-overexpressing strain than in the wild type, including minD of the Min system; cdv3 and zipN, which encode two divisome components; and murB, murC, and pbp2, which are involved in peptidoglycan (PG) synthesis. Moreover, in the rpaA-overexpressing strain, the outer membrane and cell wall PG layer were not smooth, and the outer membrane was not clearly visible by transmission electron microscopy. These results demonstrated that rpaA overexpression causes an impaired cell division, which is accompanied by transcriptional activation of cell division genes and morphological changes in the PG layer and outer membrane.
在光合微生物中,细胞周期进程取决于昼夜循环;然而,人们对细胞分裂如何响应这些环境变化进行调控却知之甚少。在单细胞球形蓝藻集胞藻PCC 6803中,RpaA参与了生物钟以及光/暗条件的信号输出。在本研究中,我们调查了双组分响应调节因子RpaA在细胞分裂调控中的作用。首先,我们检测了rpaA过表达对细胞形态以及细胞分裂相关基因表达水平的影响。通过光学显微镜观察,我们发现rpaA过表达菌株中未分裂细胞的体积增大,且分裂细胞的比例较高。在rpaA过表达菌株中,所选细胞分裂相关基因的表达水平高于野生型,包括Min系统的minD;编码两个分裂体组分的cdv3和zipN;以及参与肽聚糖(PG)合成的murB、murC和pbp2。此外,在rpaA过表达菌株中,外膜和细胞壁PG层不光滑,透射电子显微镜下外膜不清晰可见。这些结果表明,rpaA过表达导致细胞分裂受损,同时伴随着细胞分裂相关基因的转录激活以及PG层和外膜的形态变化。