Bespoke RNA recognition by Pumilios
Bespoke RNA recognition by Pumilios
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DOI：
10.1042/bst20150072
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发表时间：
2015-10-01
影响因子：
3.9
通讯作者：
Edwards, Thomas A.
中科院分区：
文献类型：
--
作者：
Edwards, Thomas A.
Pumilio 是一种最初在果蝇中发现的 RNA 结合蛋白，其 Puf 结构域由八个 Puf 重复序列组成，三个螺旋束排列成彩虹结构，其中每个重复序列识别 RNA 结合序列的单个碱基。因此，可以通过识别要改变的碱基的重复序列的突变来简单地修改八碱基识别序列，并且可以通过高分辨率晶体结构详细了解这一点。来自不同物种的多种同源物的结合机制也发生了改变，碱基从结合位点翻转以重新生成共有序列。因此，Pumilios 可以设计定制 RNA 识别序列，并可以与核酸酶融合、分裂 GFP 等，作为体外和细胞内的工具。
Pumilio is an RNA-binding protein originally identified in Drosophila, with a Puf domain made up of eight Puf repeats, three helix bundles arranged in a rainbow architecture, where each repeat recognizes a single base of the RNA-binding sequence. The eight-base recognition sequence can therefore be modified simply via mutation of the repeat that recognizes the base to be changed and this is understood in detail via high-resolution crystal structures. The binding mechanism is also altered in a variety of homologues from different species, with bases flipped out from the binding site to regenerate a consensus sequence. Thus Pumilios can be designed with bespoke RNA recognition sequences and can be fused to nucleases, split GFP, etc. as tools in vitro and in cells.