A conformational switch controls hepatitis delta virus ribozyme catalysis

被引:215
作者
Ke, AL
Zhou, KH
Ding, F
Cate, JHD
Doudna, JA [1 ]
机构
[1] Univ Calif Berkeley, Dept Mol & Cell Biol, Berkeley, CA 94705 USA
[2] Univ Calif Berkeley, Dept Chem, Berkeley, CA 94705 USA
[3] Univ Calif Berkeley, Howard Hughes Med Inst, Berkeley, CA 94705 USA
基金
美国国家卫生研究院;
关键词
D O I
10.1038/nature02522
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Ribozymes enhance chemical reaction rates using many of the same catalytic strategies as protein enzymes. In the hepatitis delta virus (HDV) ribozyme, site-specific self-cleavage of the viral RNA phosphodiester backbone(1-3) requires both divalent cations and a cytidine nucleotide(4-6). General acid - base catalysis(7-12), substrate destabilization(1,13) and global and local conformational changes(14,15) have all been proposed to contribute to the ribozyme catalytic mechanism. Here we report ten crystal structures of the HDV ribozyme in its pre-cleaved state, showing that cytidine is positioned to activate the 2'-OH nucleophile in the precursor structure. This observation supports its proposed role as a general base in the reaction mechanism. Comparison of crystal structures of the ribozyme in the pre- and post-cleavage states reveals a significant conformational change in the RNA after cleavage and that a catalytically critical divalent metal ion from the active site is ejected. The HDV ribozyme has remarkable chemical similarity to protein ribonucleases and to zymogens for which conformational dynamics are integral to biological activity. This finding implies that RNA structural rearrangements control the reactivity of ribozymes and ribonucleoprotein enzymes.
引用
收藏
页码:201 / 205
页数:5
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