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人类U4/U6.U5三snRNP的分子结构

Dmitry E Agafonov¹, Berthold Kastner¹, Olexandr Dybkov¹

¹Department of Cellular Biochemistry, Max Planck Institute for Biophysical Chemistry, D-37077 Göttingen, Germany.

Science (New York, N.Y.)

|February 26, 2016

概括

这项研究揭示了人体三种snRNP的3D结构, 该结构解释了 Brr2 酶是如何定位的,以防止过早的RNA解,这对结合体组装至关重要.

科学领域:

分子生物学
结构生物学
生物化学

背景情况:

结合体对基因表达至关重要,它介导了mRNA前结合.
U4/U6.U5三重小核核核蛋白 (tri-snRNP) 是结合体的一个基本组成部分.
了解tri-snRNP的结构对于阐明spliceosome组装和功能至关重要.

研究的目的:

确定人类三维RNP的高分辨率结构.
调查三-snRNP中的Brr2酶对RNA解的结构基础.
将人体的三核核核核蛋白结构与酵母中的相关结构进行比较,以了解结合体激活过程中的形状变化.

主要方法:

使用单粒子冷电子显微镜 (cryo-EM) 来获得1. 8兆的人类三snRNP的3D结构.
在EM密度图中安装了高分辨率的组件结构.
使用蛋白质交叉链接来验证模型.

主要成果:

确定了人类tri-snRNP的7安格斯特罗姆分辨率3D结构.
该模型阐明了 Brr2 RNA 螺旋酶的定位,防止过早的 U4/U6 RNA 解.
确定Sad1蛋白是Brr2在预激活状态中的可能结合物.

结论:

通过控制RNA解,确定结构提供了对结合体组合的调节的见解.
通过与酵母结构进行比较,推断出了Spliceosome激活过程中Brr2的形态变化和Prp8的重新排列.
这项工作为了解分子层面的结合体动力学和调节奠定了基础.

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