Crystal structure of a eukaryotic group II intron lariat.

Crystal structure of a eukaryotic group II intron lariat.
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DOI:
10.1038/nature13790
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发表时间:
2014-10-09
期刊:
影响因子:
64.8
通讯作者:
Toor, Navtej
Toor, Navtej
中科院分区:
综合性期刊1区
文献类型:
--
作者:
Robart, Aaron R.;Chan, Russell T.;Peters, Jessica K.;Rajashankar, Kanagalaghatta R.;Toor, Navtej

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支链RNA的形成是II族和剪接体内含子剪接反应的进化保守特征。该分支对于5 ‘剪接位点选择的保真度很重要,它由一个凸起的腺苷和内含子的5 ’端之间的2 ‘ -5 ’磷酸二酯键组成。为了深入了解这种普遍存在的分子内连锁,我们确定了3.7 Å中lariat形式的真核生物IIB组内含子的晶体结构。这表明,两种串联四环受体相互作用,η-η ‘和π-π ’,将结构域VI置于核心位置,使环键处于催化后状态。基于结构和生化数据,我们提出π-π ‘是一个动态相互作用,它介导了剪接两步之间的过渡,η-η ’起辅助作用。该结构还揭示了一个四镁离子簇参与催化和定位的5 '端。考虑到II族和核内含子之间的进化关系,这种活性位点结构很可能也存在于剪接体中。
The formation of branched lariat RNA is an evolutionarily conserved feature of splicing reactions for both group II and spliceosomal introns. The lariat is important for the fidelity of 5′ splice site selection and consists of a 2′-5′ phosphodiester bond between a bulged adenosine and the 5′ end of the intron. To gain insight into this ubiquitous intramolecular linkage, we determined the crystal structure of a eukaryotic group IIB intron in the lariat form at 3.7 Å. This revealed that two tandem tetraloop-receptor interactions, η-η’ and π-π’, place domain VI in the core to position the lariat bond in the post-catalytic state. Based on structural and biochemical data, we propose that π-π’ is a dynamic interaction that mediates the transition between the two steps of splicing, with η-η’ serving an ancillary role. The structure also reveals a four-magnesium-ion cluster involved in both catalysis and positioning of the 5′ end. Given the evolutionary relationship between group II and nuclear introns, it is likely that this active site configuration exists in the spliceosome as well.
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