State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
J Genet Genomics. 2011 Jun 20;38(6):253-9. doi: 10.1016/j.jgg.2011.05.001. Epub 2011 May 11.
Topoisomerase III (topo III), a type IA topoisomerase, is widespread in hyperthermophilic archaea. In order to interrogate the in vivo role of archaeal topo III, we constructed and characterized a topo III gene deletion mutant of Sulfolobus islandicus. The mutant was viable but grew more slowly than the wild-type strain, especially in a nutrient-poor medium. Flow cytometry analysis revealed changes of the mutant in growth cycle characteristics including an increase in proportion of cells containing either more than two genome equivalents or less than one genome equivalent in exponentially-growing cultures. As shown by fluorescence microscopy, a fraction of mutant cells in the cultures were drastically enlarged, and at least some of the enlarged cells were apparently capable of resuming cell division. The mutant also shows a different transcriptional profile from that of the wild-type strain. Our results suggest that the enzyme may serve roles in chromosomal segregation and control of the level of supercoiling in the cell.
拓扑异构酶 III(topo III)是一种广泛存在于嗜热古菌中的 I 型拓扑异构酶。为了研究古菌拓扑异构酶 III 的体内作用,我们构建并鉴定了 Sulfolobus islandicus 的拓扑异构酶 III 基因缺失突变体。该突变体具有活力,但比野生型菌株生长更缓慢,尤其是在营养贫乏的培养基中。流式细胞术分析显示突变体在生长周期特征方面发生了变化,包括在指数生长期培养物中含有两个以上基因组当量或少于一个基因组当量的细胞比例增加。荧光显微镜显示,培养物中的一部分突变体细胞明显增大,至少有一些增大的细胞显然能够恢复细胞分裂。该突变体还表现出与野生型菌株不同的转录谱。我们的结果表明,该酶可能在染色体分离和控制细胞中超螺旋水平方面发挥作用。