Yamamoto N, Droffner M L
Proc Natl Acad Sci U S A. 1985 Apr;82(7):2077-81. doi: 10.1073/pnas.82.7.2077.
We isolated mutant strains of the facultative anaerobe Salmonella typhimurium that grow either aerobically or anaerobically. Strict anaerobic mutants contained a defective DNA topoisomerase I gene (topI), while strict aerobic mutants contained a defective DNA gyrase subunit A gene (gyrA, also nalA). Topoisomerase I activity was detected in cell-free extracts of strict aerobic mutants but not of strict anaerobic mutant strains, whereas gyrase activity was detected in extracts of strict anaerobic mutants but not of strict aerobic mutants. Furthermore, extracts of wild-type cells, cultured under vigorous aerobic condition, contain topoisomerase I activity but no significant gyrase activity. In contrast, the extracts of anaerobically cultured wild-type cells contain gyrase activity but no significant topoisomerase I activity. Sucrose gradient centrifugation with ethidium bromide showed that chromosomal DNA in strict aerobic mutants and aerobically grown wild-type cells was relaxed, while the chromosomal DNA of strict anaerobic mutants and anaerobically grown wild-type cells was more supercoiled. Aerobic cultures of wild type and strict aerobic mutants produced both superoxide dismutase and catalase, whereas anaerobic cultures of wild type and strict anaerobic mutants did not. These results lead us to conclude that activity of topoisomerase I, associated with relaxation of chromosomal DNA, is necessary for expression of genes required for aerobic growth, whereas activity of gyrase, associated with supercoiling of chromosomal DNA, is necessary for expression of genes required for anaerobic growth.
我们分离出了鼠伤寒沙门氏菌这种兼性厌氧菌的突变菌株,它们能在需氧或厌氧条件下生长。严格厌氧突变体含有缺陷的DNA拓扑异构酶I基因(topI),而严格需氧突变体含有缺陷的DNA促旋酶亚基A基因(gyrA,也称为nalA)。在严格需氧突变体的无细胞提取物中检测到了拓扑异构酶I活性,但在严格厌氧突变体菌株的提取物中未检测到;而在严格厌氧突变体的提取物中检测到了促旋酶活性,但在严格需氧突变体的提取物中未检测到。此外,在剧烈需氧条件下培养的野生型细胞提取物含有拓扑异构酶I活性,但没有明显的促旋酶活性。相反,厌氧培养的野生型细胞提取物含有促旋酶活性,但没有明显的拓扑异构酶I活性。用溴化乙锭进行蔗糖梯度离心表明,严格需氧突变体和需氧生长的野生型细胞中的染色体DNA是松弛的,而严格厌氧突变体和厌氧生长的野生型细胞中的染色体DNA则超螺旋程度更高。野生型和严格需氧突变体的需氧培养物同时产生超氧化物歧化酶和过氧化氢酶,而野生型和严格厌氧突变体的厌氧培养物则不产生。这些结果使我们得出结论,与染色体DNA松弛相关的拓扑异构酶I活性是需氧生长所需基因表达所必需的,而与染色体DNA超螺旋相关的促旋酶活性是厌氧生长所需基因表达所必需的。