Unique structural and stabilizing roles for the individual pseudouridine residues in the 1920 region of Escherichia coli 23S rRNA
Unique structural and stabilizing roles for the individual pseudouridine residues in the 1920 region of Escherichia coli 23S rRNA
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DOI：
10.1093/nar/28.10.2075
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发表时间：
2000-05-15
影响因子：
14.9
通讯作者：
Chow, CS
中科院分区：
文献类型：
--
作者：
Meroueh, M;Grohar, PJ;Chow, CS
报道了 5'-O-BzH-2'-O-ACE 保护的假尿苷亚磷酰胺的合成 [BzH，二苯甲基氧基-双（三甲基甲硅烷氧基）甲硅烷基； ACE，双(2-乙酰氧基乙氧基)甲基]。亚磷酰胺的可用性允许可靠且高效地合成在茎或环区域含有单个或多个假尿苷修饰的发夹RNA。合成了代表大肠杆菌 23S rRNA 的 1920 环区域 (G(1906)-C-1924) 的 5 个 19-nt 发夹 RNA，其中假尿苷残基位于位置 1911、1915 和 1917，提供了所有 5 个 RNA 的热力学参数、圆二色光谱和 NMR 数据。总体而言，检查了假尿苷残基的三种不同结构背景与未修饰的 RNA 相比，我们的主要发现是假尿苷修饰对 RNA 稳定性和结构表现出一系列影响，具体取决于其位置。更具体地说，模型 RNA 的单链环区域中的假尿苷稍微不稳定，而茎环连接处的假尿苷则具有稳定作用。此外，当存在多个假尿苷残基时，观察到的对稳定性的影响近似相加。讨论了这些结果与 RNA 功能的可能关系。
The synthesis of a 5'-O-BzH-2'-O-ACE-protected pseudouridine phosphoramidite is reported [BzH, benzhydryloxy-bis(trimethylsilyloxy)sily; ACE, bis(2-acetoxyethoxy)methyl]. The availability of the phosphoramidite allows for reliable and efficient syntheses of hairpin RNAs containing single or multiple pseudouridine modifications in the stem or loop regions. Five 19-nt hairpin RNAs representing the 1920-loop region (G(1906)-C-1924) of Escherichia coli 23S rRNA were synthesized with pseudouridine residues located at positions 1911, 1915 and 1917, Thermodynamic parameters, circular dichroism spectra and NMR data are presented for all five RNAs, Overall, three different structural contexts for the pseudouridine residues were examined and compared with the unmodified RNA, Our main findings are that pseudouridine modifications exhibit a range of effects on RNA stability and structure, depending on their locations. More specifically, pseudouridines in the single-stranded loop regions of the model RNAs are slightly destabilizing, whereas a pseudouridine at the stem-loop junction is stabilizing. Furthermore, the observed effects on stability are approximately additive when multiple pseudouridine residues are present. The possible relationship of these results to RNA function is discussed.