Farewell A, Diez A A, DiRusso C C, Nyström T
Department of Microbiology, Lund University, Sweden.
J Bacteriol. 1996 Nov;178(22):6443-50. doi: 10.1128/jb.178.22.6443-6450.1996.
The increased expression of the uspA gene of Escherichia coli is an essential part of the cell's response to growth arrest. We demonstrate that stationary-phase activation of the uspA promoter is in part dependent on growth phase-dependent inactivation or repression of the FadR regulator. Transcription of uspA is derepressed during exponential growth in fadR null mutants or by including the fatty acid oleate in the growth medium of FadR+ cells. The results of DNA footprinting analysis show that FadR binds downstream of the uspA promoter in the noncoding region. Thus, uspA is a member of the fadR regulon. All the fad-lacZ fusions examined (fadBA, fadL, and fadD) are increasingly expressed in stationary phase with kinetics similar to that of the increased expression of uspA. In contrast, beta-galactosidase levels decrease during stationary phase in a fabA-lacZ lysogen, consistent with the role of FadR as an activator of fabA. The growth phase-dependent increased and decreased transcription of fad genes and fabA, respectively, is dependent on the status of the fadR gene. Cells carrying a mutation in the FadR gene (fadRS219N) that makes it nonderepressible exhibit a weak stationary-phase induction of uspA and fad genes. In addition, cells carrying fadRS219N survive long-term stasis poorly, indicating that FadR-dependent alterations in fatty acid metabolism are an integral and important part of the adaptation to stationary phase.
大肠杆菌uspA基因表达的增加是细胞对生长停滞反应的重要组成部分。我们证明,uspA启动子的稳定期激活部分依赖于FadR调节因子的生长阶段依赖性失活或抑制。在fadR缺失突变体的指数生长期间,或通过在FadR+细胞的生长培养基中加入脂肪酸油酸,uspA的转录被去抑制。DNA足迹分析结果表明,FadR在非编码区的uspA启动子下游结合。因此,uspA是fadR调控子的成员。所有检测的fad-lacZ融合体(fadBA、fadL和fadD)在稳定期的表达都增加,其动力学与uspA表达增加的动力学相似。相比之下,在fabA-lacZ溶原菌中,β-半乳糖苷酶水平在稳定期下降,这与FadR作为fabA激活剂的作用一致。fad基因和fabA的生长阶段依赖性转录增加和减少分别取决于fadR基因的状态。携带FadR基因(fadRS219N)突变使其不可抑制的细胞,uspA和fad基因的稳定期诱导较弱。此外,携带fadRS219N的细胞在长期停滞中存活能力较差,这表明脂肪酸代谢中依赖FadR的改变是适应稳定期不可或缺的重要部分。