Molecular Determinants for 23S rRNA Recognition and Modification by the E-coli Pseudouridine Synthase RIuE

JOURNAL OF MOLECULAR BIOLOGY

卷:430

Article

Tillault, Anne-Sophie¹  Schultz, Sarah K.¹  Wieden, Hans-Joachim¹  Kothe, Ute¹ 

[1] Univ Lethbridge, Alberta RNA Res & Training Inst, Dept Chem & Biochem, Lethbridge, AB T1K 3M4, Canada

关键词: ribosomal RNA;    peptidyltransferase center;    RNA modification;    pseudouridine;    RNA-protein interaction;   

DOI  :  10.1016/j.jmb.2018.03.011

来源: Elsevier

【 摘 要 】

The isomerization of uridine to pseudouridine is the most common type of RNA modification found in RNAs across all domains of life and is performed by RNA-dependent and RNA-independent enzymes. The Escherichia coli pseudouridine synthase RIuE acts as a stand-alone, highly specific enzyme forming the universally conserved pseudouridine at position 2457, located in helix 89 (H89) of the 23S rRNA in the peptidyltransferase center. Here, we conduct a detailed structure function analysis to determine the structural elements both in RIuE and in 23S rRNA required for RNA protein interaction and pseudouridine formation. We determined that RIuE recognizes a large part of 23S rRNA comprising both H89 and the single-stranded flanking regions which explains the high substrate specificity of RIuE. Within RIuE, the target RNA is recognized through sequence-specific contacts with loop L7-8 as well as interactions with loop L1-2 and the flexible N-terminal region. We demonstrate that RIuE is a faster pseudouridine synthase than other enzymes which likely enables it to act in the early stages of ribosome formation. In summary, our biochemical characterization of RIuE provides detailed insight into the molecular mechanism of RIuE forming a highly conserved pseudouridine during ribosome biogenesis. (C) 2018 The Authors. Published by Elsevier Ltd.

【 授权许可】

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