柯萨奇病毒B3三叶草叶形RNA茎环D亚结构域的结构及其与蛋白酶3C的相互作用。

The structure of the stemloop D subdomain of coxsackievirus B3 cloverleaf RNA and its interaction with the proteinase 3C.

作者信息

Ohlenschläger Oliver, Wöhnert Jens, Bucci Enrico, Seitz Simone, Häfner Sabine, Ramachandran Ramadurai, Zell Roland, Görlach Matthias

机构信息

Institut für Molekulare Biotechnologie eV, Bentenbergstr 100813, D-07745 Jena, Germany.

出版信息

Structure. 2004 Feb;12(2):237-48. doi: 10.1016/j.str.2004.01.014.

DOI:10.1016/j.str.2004.01.014

PMID:14962384

Abstract

Stemloop D (SLD) of the 5' cloverleaf RNA is the cognate ligand of the coxsackievirus B3 (CVB3) 3C proteinase (3Cpro). Both are indispensable components of the viral replication initiation complex. SLD is a structurally autonomous subunit of the 5' cloverleaf. The SLD structure was solved by NMR spectroscopy to an rms deviation of 0.66 A (all heavy atoms). SLD contains a novel triple pyrimidine mismatch motif with a central Watson-Crick type C:U pair. SLD is capped by an apical uCACGg tetraloop adopting a structure highly similar to stable cUNCGg tetraloops. Binding of CVB3 3Cpro induces changes in NMR spectra for nucleotides adjacent to the triple pyrimidine mismatch and of the tetraloop implying them as sites of specific SLD:3Cpro interaction. The binding of 3Cpro to SLD requires the integrity of those structural elements, strongly suggesting that 3Cpro recognizes a structural motif instead of a specific sequence.

摘要

5' 三叶草RNA的茎环D（SLD）是柯萨奇病毒B3（CVB3）3C蛋白酶（3Cpro）的同源配体。两者都是病毒复制起始复合物的不可或缺的组成部分。SLD是5' 三叶草的一个结构自主亚基。通过核磁共振光谱法解析了SLD的结构，其均方根偏差为0.66 Å（所有重原子）。SLD包含一个新型的三联嘧啶错配基序，中心为沃森-克里克型C:U碱基对。SLD由顶端的uCACGg四环封闭，其结构与稳定的cUNCGg四环高度相似。CVB3 3Cpro的结合会导致三联嘧啶错配附近核苷酸以及四环的核磁共振光谱发生变化，这意味着它们是SLD与3Cpro特异性相互作用的位点。3Cpro与SLD的结合需要这些结构元件的完整性，这强烈表明3Cpro识别的是一个结构基序而非特定序列。