Urios A, Herrera G, Aleixandre V, Blanco M
Instituto de Investigaciones Citológicas de la Caja de Ahorros de Valencia, Spain.
Mutat Res. 1990 Apr;243(4):267-72. doi: 10.1016/0165-7992(90)90142-7.
We studied the influence of DNA topological changes on Escherichia coli recA gene expression. This was monitored by measuring beta-galactosidase activity in cells containing a recA-lacZ fusion. To modulate DNA supercoiling we used mutations in the genes encoding for topoisomerase I and DNA gyrase. After either UV irradiation or treatment with the gyrase inhibitor ciprofloxacin, induction of the recA gene was reduced in topA10 mutants, this reduction being alleviated when gyrA or gyrB mutations causing DNA relaxation were present. A reduced induction of recA was also observed after incubation of cells carrying the recA441 mutation at 42 degrees C in the presence of adenine. Using bacteria deficient in the LexA repressor, we have demonstrated that the topA10 mutation reduces the constitutive expression of the recA gene. We suggest that the increase in negative supercoiling resulting from topoisomerase I deficiency interferes with transcription from the recA promoter. The reduction in the expression of the recA gene in topA10 bacteria could determine their increased UV sensitivity as well as their partial defectiveness in SOS mutability.
我们研究了DNA拓扑结构变化对大肠杆菌recA基因表达的影响。通过测量含有recA - lacZ融合基因的细胞中的β-半乳糖苷酶活性来监测这种影响。为了调节DNA超螺旋,我们利用了编码拓扑异构酶I和DNA促旋酶的基因突变。在紫外线照射或用促旋酶抑制剂环丙沙星处理后,topA10突变体中recA基因的诱导表达降低,当存在导致DNA松弛的gyrA或gyrB突变时,这种降低得到缓解。在携带recA441突变的细胞于42℃在腺嘌呤存在下培养后,也观察到recA的诱导表达降低。利用缺乏LexA阻遏物的细菌,我们证明topA10突变降低了recA基因的组成型表达。我们认为,拓扑异构酶I缺乏导致的负超螺旋增加会干扰recA启动子的转录。topA10细菌中recA基因表达的降低可能决定了它们对紫外线敏感性的增加以及它们在SOS诱变中的部分缺陷。