Lu Zuolei, Takeuchi Michio, Sato Tsutomu
Department of Applied Biological Science, Tokyo University of Agriculture and Technology, Fuchu, Tokyo 183-8509, Japan.
J Bacteriol. 2007 Aug;189(15):5642-51. doi: 10.1128/JB.00467-07. Epub 2007 May 25.
We have carried out a functional analysis of LysR family transcriptional regulators in Bacillus subtilis. The cell density of cultures of a yofA insertion mutant declined sharply after the end of exponential growth, as measured by optical density at 600 nm. Complementation in trans and analysis of isopropyl-beta-d-thiogalactopyranoside (IPTG)-dependent growth of an inducible yofA strain confirmed that YofA contributes to the cell density of a culture after the end of exponential growth. Microscopic observation suggested that cell division is inhibited or delayed in the yofA mutant during entry into stationary phase. Analysis of the transcription of cell division genes revealed that the expression of ftsW is inhibited in yofA mutants, and overexpression of yofA, driven by a multiple-copy plasmid, enhances the induction of ftsW expression. These results suggest that YofA is required for the final round of cell division before entry into stationary phase and that YofA positively regulates ftsW expression. The defects caused by mutation of yofA were suppressed in strains carrying P(spac)-ftsW in the presence of IPTG. Furthermore, maximal expression of yofA was observed at the onset of stationary phase, which coincided with the maximal ftsW expression. Our data indicate that YofA is involved in cell division through positive regulation of the expression of ftsW in B. subtilis.
我们对枯草芽孢杆菌中的LysR家族转录调节因子进行了功能分析。通过在600nm处的光密度测量,yofA插入突变体培养物的细胞密度在指数生长结束后急剧下降。反式互补以及对可诱导yofA菌株的异丙基-β-D-硫代半乳糖苷(IPTG)依赖性生长的分析证实,YofA在指数生长结束后对培养物的细胞密度有贡献。显微镜观察表明,yofA突变体在进入稳定期时细胞分裂受到抑制或延迟。对细胞分裂基因转录的分析表明,yofA突变体中ftsW的表达受到抑制,由多拷贝质粒驱动的yofA过表达增强了ftsW表达的诱导。这些结果表明,YofA是进入稳定期前最后一轮细胞分裂所必需的,并且YofA正向调节ftsW的表达。在IPTG存在的情况下,携带P(spac)-ftsW的菌株中yofA突变引起的缺陷得到了抑制。此外,在稳定期开始时观察到yofA的最大表达,这与ftsW的最大表达一致。我们的数据表明,YofA通过正向调节枯草芽孢杆菌中ftsW的表达参与细胞分裂。