Nesbø Camilla L, Doolittle W Ford
Department of Biochemistry and Molecular Biology, Dalhousie University, Halifax, NS, Canada B3H 1X5.
Proc Natl Acad Sci U S A. 2003 Sep 16;100(19):10806-11. doi: 10.1073/pnas.1434268100. Epub 2003 Aug 28.
Group I introns are common in the 23 rRNA genes of mitochondria and chloroplasts. Often, they encode "homing endonucleases," which target highly conserved gene sequences and drive interorganellar intron mobility, even across species and genus lines. Most bacterial 23S rRNA genes show these same endonuclease-sensitive target sequences. However, only two bacterial 23S rRNA genes are known to contain group I introns: that of Simkania negevensis [Everett, K. D., Kahane, S., Bush, R. M. & Friedman, M. G. (1999) J. Bacteriol. 181, 4734-4740], where the intron is not spliced and probably limits growth, and that of Coxiella burnetii [Seshadri, R., et al. (2003) Proc. Natl. Acad. Sci. USA 100, 5455-5460], where no direct evidence of splicing exists. Both bacteria are intracellular parasites and might have acquired introns from eukaryotic hosts. Here we provide direct evidence for splicing, and evolutionary evidence for mobility, of group I introns in the 23S rRNA genes of several free-living hyperthermophilic bacteria of the genus Thermotoga. These bacteria do not live closely with eukaryotes, but phylogenetic analyses suggest that their introns were also acquired from eukaryotic (probably algal) organelles. In vivo, their introns must be spliced at temperatures approaching 90 degrees C, making them the most thermostable natural ribozymes so far described. We demonstrate that at least some of these introns can also self-splice in vitro.
I 类内含子常见于线粒体和叶绿体的 23 rRNA 基因中。它们通常编码“归巢内切核酸酶”,这些酶靶向高度保守的基因序列,并驱动细胞器间内含子的移动,甚至跨越物种和属的界限。大多数细菌的 23S rRNA 基因都显示出这些相同的内切核酸酶敏感靶序列。然而,已知只有两个细菌的 23S rRNA 基因含有 I 类内含子:嗜肺军团菌(Simkania negevensis)的基因 [埃弗雷特,K. D.,卡哈内,S.,布什,R. M. & 弗里德曼,M. G.(1999 年)《细菌学杂志》181 卷,4734 - 4740 页],其中的内含子不进行剪接,可能会限制生长;以及贝氏考克斯氏体(Coxiella burnetii)的基因 [塞沙德里,R. 等人(2003 年)《美国国家科学院院刊》100 卷,5455 - 5460 页],目前尚无剪接的直接证据。这两种细菌都是细胞内寄生虫,可能从真核宿主那里获得了内含子。在此,我们提供了直接证据,证明嗜热栖热菌属(Thermotoga)几种自由生活的嗜热细菌的 23S rRNA 基因中的 I 类内含子存在剪接现象,以及其移动性的进化证据。这些细菌并不与真核生物紧密共生,但系统发育分析表明它们的内含子也是从真核(可能是藻类)细胞器获得的。在体内,它们的内含子必须在接近 90 摄氏度的温度下进行剪接,这使它们成为迄今为止所描述的最耐热的天然核酶。我们证明,这些内含子中至少有一些在体外也能自我剪接。