Jovanovich S B, Martinell M, Record M T, Burgess R R
McArdle Laboratory for Cancer Research, Madison, Wisconsin 53706.
J Bacteriol. 1988 Feb;170(2):534-9. doi: 10.1128/jb.170.2.534-539.1988.
The rapid in vivo response of both Escherichia coli and Salmonella typhimurium osmoregulated genes to an osmotic upshift was analyzed in detail by using chromosomal operon fusions. Within 10 min after the addition of 0.3 M NaCl to the culture medium, the differential rates of expression of both an S. typhimurium proU-lac fusion and a proP-lac fusion increased by 180- and 17-fold respectively, while an E. coli ompC-lac fusion increased by 3.4-fold. For all three stimulated promoters, the increased rate of expression was maintained until about 40 min after the osmotic upshift. Thereafter, proU expression continued at a steady-state rate that was 27-fold higher than that of the control, while proP and ompC expression fell to 1.4- and 2-fold of the control rates, respectively. In contrast, expression of an E. coli ompF-lac fusion decreased twofold within 2.5 min. For proU, the length of the lag phase, which preceded the onset of the rapid response, increased with the degree of osmotic upshift, above a threshold of 0.2 M NaCl; the onset of the rapid proU response also preceded the resumption of growth. The rapid response phase, which was first quantitated for proU, proP, ompC, and ompF in this study, is an important component of the osmoregulation of these promoters. The addition of the osmoprotectant glycine betaine at the time of osmotic upshift decreased both the length of the rapid response and the subsequent steady-state of expression of proU.
利用染色体操纵子融合技术,详细分析了大肠杆菌和鼠伤寒沙门氏菌渗透调节基因在体内对渗透压升高的快速反应。向培养基中添加0.3M NaCl后10分钟内,鼠伤寒沙门氏菌proU-lac融合体和proP-lac融合体的差异表达率分别增加了180倍和17倍,而大肠杆菌ompC-lac融合体增加了3.4倍。对于所有三个受刺激的启动子,表达增加率一直维持到渗透压升高后约40分钟。此后,proU表达以稳定状态持续,比对照高27倍,而proP和ompC表达分别降至对照率的1.4倍和2倍。相比之下,大肠杆菌ompF-lac融合体的表达在2.5分钟内下降了两倍。对于proU,在快速反应开始之前的延迟期长度随着渗透压升高程度的增加而增加,高于0.2M NaCl的阈值;proU的快速反应开始也先于生长的恢复。本研究中首次对proU、proP、ompC和ompF进行定量的快速反应阶段,是这些启动子渗透调节的重要组成部分。在渗透压升高时添加渗透保护剂甘氨酸甜菜碱,可缩短快速反应的长度,并降低proU随后的表达稳态。