Structure-based insights into self-cleavage by a four-way junctional twister-sister ribozyme.

Structure-based insights into self-cleavage by a four-way junctional twister-sister ribozyme.
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基于结构的四路连接扭转姐妹核酶自裂解的见解

DOI:
10.1038/s41467-017-01276-y
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发表时间:
2017-10-30
影响因子:
16.6
通讯作者:
Ren A
Ren A
中科院分区:
综合性期刊1区
文献类型:
--
作者:
Zheng L;Mairhofer E;Teplova M;Zhang Y;Ma J;Patel DJ;Micura R;Ren A

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在这里,我们报告的晶体结构和切割试验的四路连接扭转姐妹自切割核酶。值得注意的是,11个保守的空间上分离的环核苷酸被带到接近的核酶核心通过长距离的相互作用介导的水合镁2+阳离子。裂解位点处的C62-A63步骤采用分开的方向,柔性C62向外定向,而A63向内定向并通过堆叠和氢键相互作用锚定。结构指导的研究关键基地,糖,磷酸盐突变的扭转姐妹核酶,建议贡献的切割化学之间的相互作用鸟嘌呤在活性位点和非桥接氧的易裂磷酸盐,以前发现的功能也为相关的扭转核酶。我们的四路交界前催化结构显着不同的对齐在切割步骤(splayed-apart与基地堆叠)和周围的残基和水合Mg 2+离子相对于报告的三路交界前催化结构的扭转姐妹核酶。
Here we report on the crystal structure and cleavage assays of a four-way junctional twister-sister self-cleaving ribozyme. Notably, 11 conserved spatially separated loop nucleotides are brought into close proximity at the ribozyme core through long-range interactions mediated by hydrated Mg2+cations. The C62–A63 step at the cleavage site adopts a splayed-apart orientation, with flexible C62 directed outwards, whereas A63 is directed inwards and anchored by stacking and hydrogen-bonding interactions. Structure-guided studies of key base, sugar, and phosphate mutations in the twister-sister ribozyme, suggest contributions to the cleavage chemistry from interactions between a guanine at the active site and the non-bridging oxygen of the scissile phosphate, a feature found previously also for the related twister ribozyme. Our four-way junctional pre-catalytic structure differs significantly in the alignment at the cleavage step (splayed-apart vs. base-stacked) and surrounding residues and hydrated Mg2+ions relative to a reported three-way junctional pre-catalytic structure of the twister-sister ribozyme.
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发表时间: 2017-03
期刊: RNA (New York, N.Y.)
影响因子: --
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发表时间: 2014-09-09
影响因子: 11.1
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