Boyle D S, Khattar M M, Addinall S G, Lutkenhaus J, Donachie W D
Institute of Cell and Molecular Biology, University of Edinburgh, UK.
Mol Microbiol. 1997 Jun;24(6):1263-73. doi: 10.1046/j.1365-2958.1997.4091773.x.
In the absence of exogenous promoters, plasmid-mediated complementation of the temperature-sensitive ftsW201 allele requires the presence of the full coding sequence of ftsW plus upstream DNA encompassing the C-terminus of mraY and the full coding sequence of murD. We used molecular and genetic techniques to introduce an insertional inactivation into the chromosomal copy of ftsW, in the presence of the plasmid-borne wild-type ftsW gene under the control of P(BAD). In the absence of arabinose, the ftsW-null strain is not viable, and a shift from arabinose- to glucose-containing liquid medium resulted in a block in division, followed by cell lysis. Immunofluorescence microscopy revealed that in ftsW-null filaments, the FtsZ ring is absent in 50-60% of filaments, whilst between one and three Z-rings per filament can be detected in the remainder of the population, with the majority of these containing only one Z-ring per filament. We also demonstrated that the expression of only ftsWS (the smaller of two ftsW open reading frames) from P(BAD) is sufficient for complementation of the ftsW-null allele. We conclude that FtsW is an essential cell-division protein in Escherichia coli, and that it plays a role in the stabilization of the FtsZ ring during cell division.
在没有外源启动子的情况下,质粒介导的对温度敏感的ftsW201等位基因的互补作用需要ftsW完整编码序列以及包含mraY C末端的上游DNA和murD完整编码序列的存在。我们使用分子和遗传技术,在受P(BAD)控制的质粒携带的野生型ftsW基因存在的情况下,对ftsW的染色体拷贝进行插入失活。在没有阿拉伯糖的情况下,ftsW缺失菌株无法存活,从含阿拉伯糖的液体培养基转换为含葡萄糖的液体培养基会导致分裂受阻,随后细胞裂解。免疫荧光显微镜检查显示,在ftsW缺失的丝状体中,50%-60%的丝状体中不存在FtsZ环,而在其余群体中,每根丝状体可检测到一到三个Z环,其中大多数每根丝状体仅含有一个Z环。我们还证明,仅从P(BAD)表达ftsWS(ftsW两个开放阅读框中较小的一个)就足以互补ftsW缺失等位基因。我们得出结论,FtsW是大肠杆菌中一种必需的细胞分裂蛋白,并且它在细胞分裂过程中FtsZ环的稳定中发挥作用。