Slomińska M, Neubauer P, Wegrzyn G
Department of Molecular Biology, University of Gdańsk, Kladki 24, Gdańsk, 80-822, Poland.
Virology. 1999 Sep 30;262(2):431-41. doi: 10.1006/viro.1999.9907.
On infection of its host, Escherichia coli, bacteriophage lambda can follow one of two alternative developmental pathways: lytic or lysogenic. Here we demonstrate that the "lysis-versus-lysogenization" decision is influenced by guanosine tetraphosphate (ppGpp), a nucleotide that is synthesized in E. coli cells in response to amino acid or carbon source starvation. We found that the efficiency of lysogenization is the highest at ppGpp concentrations somewhat higher than the basal level; too low and too high levels of ppGpp result in less efficient lysogenization. Maintenance of the already integrated lambda prophage and phage lytic development were not significantly influenced in the host lacking ppGpp. We found that the level of HflB/FtsH protease, responsible for degradation of the CII protein, an activator of "lysogenic" promoters, depends on ppGpp concentration. The highest levels of HflB/FtsH was found in bacteria lacking ppGpp and in cells bearing increased concentrations of this nucleotide. Using lacZ fusions, we investigated the influence of ppGpp on activities of lambda promoters important at the stage of the lysis-versus-lysogenization decision. We found that each promoter is regulated differentially in response to the abundance of ppGpp. Moreover, our results suggest that the cAMP level may influence ppGpp concentration in cells. The mechanism of the ppGpp-mediated control of lambda development at the stage of the lysis-versus-lysogenization decision may be explained on the basis of differential influence of guanosine tetraphosphate on activities of p(L), p(R), p(E), p(I), and p(aQ) promoters and by dependence of HflB/FtsH protease level on ppGpp concentration.
在感染其宿主大肠杆菌时,噬菌体λ可以遵循两种不同的发育途径之一:裂解或溶原。在这里,我们证明“裂解与溶原化”的决定受四磷酸鸟苷(ppGpp)的影响,ppGpp是一种在大肠杆菌细胞中响应氨基酸或碳源饥饿而合成的核苷酸。我们发现,在ppGpp浓度略高于基础水平时,溶原化效率最高;ppGpp水平过低和过高都会导致溶原化效率降低。在缺乏ppGpp的宿主中,已整合的λ原噬菌体的维持和噬菌体裂解发育没有受到显著影响。我们发现,负责降解“溶原”启动子激活剂CII蛋白的HflB/FtsH蛋白酶水平取决于ppGpp浓度。在缺乏ppGpp的细菌和这种核苷酸浓度增加的细胞中发现了最高水平的HflB/FtsH。使用lacZ融合,我们研究了ppGpp对在裂解与溶原化决定阶段重要的λ启动子活性的影响。我们发现每个启动子对ppGpp丰度的反应都有差异调节。此外,我们的结果表明cAMP水平可能影响细胞中的ppGpp浓度。ppGpp在裂解与溶原化决定阶段对λ发育的介导控制机制可以基于四磷酸鸟苷对p(L)、p(R)、p(E)、p(I)和p(aQ)启动子活性的差异影响以及HflB/FtsH蛋白酶水平对ppGpp浓度的依赖性来解释。