Szalewska-Pałasz A, Wegrzyn G
Department of Molecular Biology, University of Gdańsk, Poland.
Acta Biochim Pol. 1995;42(2):233-9.
Replication of lambda plasmid DNA is halted in amino acid-starved wild type (stringent) strains whereas it proceeds in relA (relaxed) mutants. The only transcription which could be important in lambda plasmid DNA replication in amino acid-starved Escherichia coli cells is that starting from the pR promoter. Using a fusion which consists of the lacZ gene under the control of bacteriophage lambda pR promoter we found that transcription starting from this promoter was inhibited during the stringent, but not the relaxed, response in E. coli. We confirmed our conclusion by estimating the relative level of the pR transcript by RNA-DNA hybridization. We propose that decreased transcription from the pR promoter which serves as transcriptional activation of ori lambda is responsible for inhibition of lambda plasmid replication during the stringent response. The results presented in this paper, combined with our recent findings (published elsewhere), indicate that the transcriptional activation of ori lambda may be a main regulatory process controlling lambda DNA replication not only during the relaxed response but also in normal growth conditions.
在氨基酸饥饿的野生型(严谨型)菌株中,λ质粒DNA的复制会停止,而在relA(松弛型)突变体中则会继续进行。在氨基酸饥饿的大肠杆菌细胞中,唯一可能对λ质粒DNA复制重要的转录是从pR启动子开始的转录。使用一个由噬菌体λ pR启动子控制的lacZ基因组成的融合体,我们发现从这个启动子开始的转录在大肠杆菌的严谨反应期间受到抑制,但在松弛反应期间不受抑制。我们通过RNA-DNA杂交估计pR转录本的相对水平来证实了我们的结论。我们提出,作为ori λ转录激活的pR启动子转录减少是严谨反应期间λ质粒复制受到抑制的原因。本文给出的结果,结合我们最近的发现(已在其他地方发表),表明ori λ的转录激活可能是不仅在松弛反应期间而且在正常生长条件下控制λ DNA复制的主要调节过程。