Asai T, Zaporojets D, Squires C, Squires C L
Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, MA 02111, USA.
Proc Natl Acad Sci U S A. 1999 Mar 2;96(5):1971-6. doi: 10.1073/pnas.96.5.1971.
Current global phylogenies are built predominantly on rRNA sequences. However, an experimental system for studying the evolution of rRNA is not readily available, mainly because the rRNA genes are highly repeated in most experimental organisms. We have constructed an Escherichia coli strain in which all seven chromosomal rRNA operons are inactivated by deletions spanning the 16S and 23S coding regions. A single E. coli rRNA operon carried by a multicopy plasmid supplies 16S and 23S rRNA to the cell. By using this strain we have succeeded in creating microorganisms that contain only a foreign rRNA operon derived from either Salmonella typhimurium or Proteus vulgaris, microorganisms that have diverged from E. coli about 120-350 million years ago. We also were able to replace the E. coli rRNA operon with an E. coli/yeast hybrid one in which the GTPase center of E. coli 23S rRNA had been substituted by the corresponding domain from Saccharomyces cerevisiae. These results suggest that, contrary to common belief, coevolution of rRNA with many other components in the translational machinery may not completely preclude the horizontal transfer of rRNA genes.
当前的全球系统发育树主要基于rRNA序列构建。然而,用于研究rRNA进化的实验系统并不容易获得,主要原因是rRNA基因在大多数实验生物中高度重复。我们构建了一种大肠杆菌菌株,其中所有七个染色体rRNA操纵子通过跨越16S和23S编码区域的缺失而失活。由多拷贝质粒携带的单个大肠杆菌rRNA操纵子为细胞提供16S和23S rRNA。通过使用这种菌株,我们成功地创造了仅含有源自鼠伤寒沙门氏菌或普通变形杆菌的外源rRNA操纵子的微生物,这些微生物与大肠杆菌在大约1.2亿至3.5亿年前就已经分化。我们还能够用大肠杆菌/酵母杂交操纵子取代大肠杆菌rRNA操纵子,其中大肠杆菌23S rRNA的GTPase中心已被酿酒酵母的相应结构域取代。这些结果表明,与普遍看法相反,rRNA与翻译机制中许多其他成分的共同进化可能并不完全排除rRNA基因的水平转移。