IMCB University of Tartu, Riia 23, 51010 Tartu, Estonia.
Nucleic Acids Res. 2024 Jun 24;52(11):6614-6628. doi: 10.1093/nar/gkae222.
Ribosomal RNA modifications are introduced by specific enzymes during ribosome assembly in bacteria. Deletion of individual modification enzymes has a minor effect on bacterial growth, ribosome biogenesis, and translation, which has complicated the definition of the function of the enzymes and their products. We have constructed an Escherichia coli strain lacking 10 genes encoding enzymes that modify 23S rRNA around the peptidyl-transferase center. This strain exhibits severely compromised growth and ribosome assembly, especially at lower temperatures. Re-introduction of the individual modification enzymes allows for the definition of their functions. The results demonstrate that in addition to previously known RlmE, also RlmB, RlmKL, RlmN and RluC facilitate large ribosome subunit assembly. RlmB and RlmKL have functions in ribosome assembly independent of their modification activities. While the assembly stage specificity of rRNA modification enzymes is well established, this study demonstrates that there is a mutual interdependence between the rRNA modification process and large ribosome subunit assembly.
核糖体 RNA 修饰是由细菌中核糖体组装过程中的特定酶引入的。单个修饰酶的缺失对细菌生长、核糖体生物发生和翻译的影响较小,这使得酶及其产物的功能定义变得复杂。我们构建了一种大肠杆菌菌株,该菌株缺乏 10 个编码修饰位于肽基转移酶中心周围的 23S rRNA 的酶的基因。该菌株表现出严重的生长和核糖体组装受损,特别是在较低温度下。单独修饰酶的重新引入可以定义其功能。结果表明,除了先前已知的 RlmE 外,RlmB、RlmKL、RlmN 和 RluC 也有助于大亚基核糖体的组装。RlmB 和 RlmKL 的核糖体组装功能与其修饰活性无关。虽然 rRNA 修饰酶的组装阶段特异性已经得到很好的证实,但本研究表明 rRNA 修饰过程和大亚基核糖体组装之间存在相互依存关系。