Balke V L, Gralla J D
Department of Chemistry and Biochemistry, University of California, Los Angeles 90024.
J Bacteriol. 1987 Oct;169(10):4499-506. doi: 10.1128/jb.169.10.4499-4506.1987.
The supercoiling levels of plasmid DNA were determined from Escherichia coli which was grown in ways that are known to alter global patterns of gene expression and metabolism. Changes in DNA supercoiling were shown to occur during several types of these nutrient upshifts and downshifts. The most dramatic change in supercoiling was seen in starved cells, in which two populations of differentially relaxed plasmids were shown to coexist. Thus, some changes in the external nutritional environment that cause the cells to reorganize their global metabolism also cause accompanying changes in DNA supercoiling. Results of experiments with dinitrophenol suggested that the observed relaxations were probably not due to reduced pools of ATP. When rifampin was used to release supercoils restrained by RNA polymerase, the cellular topoisomerases responded by removing these new, unrestrained supercoils. We interpret these results as implying that the cellular topological machinery maintains a constant superhelical energy in the DNA except during certain growth transitions, when changes in metabolism and gene expression are accompanied by changes in DNA supercoiling.
从以已知会改变基因表达和代谢全局模式的方式生长的大肠杆菌中测定质粒DNA的超螺旋水平。在几种类型的营养物质上调和下调过程中,DNA超螺旋发生了变化。在饥饿细胞中观察到超螺旋的最显著变化,其中显示有两种不同程度松弛的质粒群体共存。因此,一些导致细胞重新组织其全局代谢的外部营养环境变化也会引起DNA超螺旋的相应变化。二硝基苯酚的实验结果表明,观察到的松弛可能不是由于ATP池减少所致。当使用利福平释放受RNA聚合酶限制的超螺旋时,细胞拓扑异构酶会通过去除这些新的、不受限制的超螺旋做出反应。我们将这些结果解释为意味着细胞拓扑机制在DNA中维持恒定的超螺旋能量,除非在某些生长转变期间,此时代谢和基因表达的变化伴随着DNA超螺旋的变化。