A protein-based EM label for RNA identifies the location of exons in spliceosomes
A protein-based EM label for RNA identifies the location of exons in spliceosomes
复制标题
DOI：
10.1038/nsmb.1378
复制
发表时间：
2008-02-01
影响因子：
16.8
通讯作者：
Jurica, Melissa S.
中科院分区：
文献类型：
--
作者：
Alcid, Eric A.;Jurica, Melissa S.
为了通过EM定位剪接体结构中的关键RNA特征，我们将序列特异性RNA结合蛋白与具有独特甜甜圈状结构的蛋白融合。我们使用这种融合来标记剪接体，剪接体组装在外显子中含有靶序列的前体mRNA上。该标签在剪接体的EM图像中清晰可见，随后的平均图像处理显示外显子在复合体中彼此靠近。这种标记策略将作为分析含RNA大分子复合物结构的通用工具。
To locate key RNA features in the structure of the spliceosome by EM, we fused a sequence-specific RNA binding protein to a protein with a distinct donut-shaped structure. We used this fusion to label spliceosomes assembled on a pre-mRNA that contained the target sequence in the exons. The label is clearly visible in EM images of the spliceosome, and subsequent image processing with averaging shows that the exons sit close to each other in the complex. This labeling strategy will serve as a general tool for analyzing the structures of RNA-containing macromolecular complexes.