Popova Anna M, Jain Nikhil, Dong Xiyu, Abdollah-Nia Farshad, Britton Robert A, Williamson James R
Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, TX 77030, USA, Center for Metagenomics and Microbiome Research, Baylor College of Medicine, Houston, TX 77030, USA.
bioRxiv. 2024 May 11:2024.05.10.593627. doi: 10.1101/2024.05.10.593627.
Ribosomal RNA modifications in prokaryotes have been sporadically studied, but there is a lack of a comprehensive picture of modification sites across bacterial phylogeny. is a preeminent model organism for gram-positive bacteria, with a well-annotated and editable genome, convenient for fundamental studies and industrial use. Yet remarkably, there has been no complete characterization of its rRNA modification inventory. By expanding modern MS tools for the discovery of RNA modifications, we found a total of 25 modification sites in 16S and 23S rRNA of including the chemical identity of the modified nucleosides and their precise sequence location. Furthermore, by perturbing large subunit biogenesis using depletion of an essential factor RbgA and measuring the completion of 23S modifications in the accumulated intermediate, we provide a first look at the order of modification steps during the late stages of assembly in . While our work expands the knowledge of bacterial rRNA modification patterns, adding to the list of fully annotated species after and in a broader context, it provides the experimental framework for discovery and functional profiling of rRNA modifications to ultimately elucidate their role in ribosome biogenesis and translation.
原核生物中的核糖体RNA修饰已得到零星研究，但缺乏对细菌系统发育中修饰位点的全面了解。 是革兰氏阳性菌的卓越模式生物，具有注释完善且可编辑的基因组，便于进行基础研究和工业应用。然而，值得注意的是，其rRNA修饰清单尚未得到完整表征。通过扩展用于发现RNA修饰的现代质谱工具，我们在 的16S和23S rRNA中总共发现了25个修饰位点，包括修饰核苷的化学特性及其精确的序列位置。此外，通过利用必需因子RbgA的缺失干扰大亚基生物合成，并测量积累中间体中23S修饰的完成情况，我们首次了解了 在组装后期修饰步骤的顺序。虽然我们的工作扩展了对细菌rRNA修饰模式的认识，在更广泛的背景下将 添加到经过全面注释的物种列表中，但它为rRNA修饰的发现和功能分析提供了实验框架，以最终阐明它们在核糖体生物合成和翻译中的作用。
