Sequence-specific RNA binding mediated by the RNase PH domain of components of the exosome
Sequence-specific RNA binding mediated by the RNase PH domain of components of the exosome
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DOI：
10.1261/rna.144606
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发表时间：
2006-10-01
期刊：
影响因子：
4.5
通讯作者：
Wilusz, Jeffrey
中科院分区：
文献类型：
--
作者：
Anderson, John R.;Mukherjee, Devi;Wilusz, Jeffrey
我们先前已经证明，PM-SCL-75是人类外体复合体的一种成分，参与RNA的成熟和mRNA的衰退，可以与含有富含AU的不稳定元件的RNA特异地相互作用。通过对一系列缺失突变体的分析，我们现在已经表明，代表RNase PH结构域的266个氨基酸片段负责与富含AU的元素的序列特异性结合。此外，我们发现来自另外两个外体成分OIP2和RRP41的RNase PH结构域以及来自大肠杆菌多核苷酸磷酸化酶的RNase PH结构域都能够与含有类似亲和力的富含AU元件的RNA特异性地相互作用。最后，我们证明PM-SCL-75的RNase PH结构域的相互作用很容易被聚(U)竞争，但只有在其他同聚RNA的作用下才有效。这些数据表明，RNase PH结构域通常对富含U和AU的序列具有亲和力，并拓宽了包含这些结构域的蛋白质在RNA生物学中的潜在作用。
We have previously demonstrated that PM-Scl-75, a component of the human exosome complex involved in RNA maturation and mRNA decay, can specifically interact with RNAs containing an AU-rich instability element. Through the analysis of a series of deletion mutants, we have now shown that a 266 amino acid fragment representing the RNase PH domain is responsible for the sequence-specific binding to AU-rich elements. Furthermore, we found that the RNase PH domains from two other exosomal components, OIP2 and RRP41, as well as from Escherichia coli polynucleotide phosphorylase, are all capable of specifically interacting with RNAs containing an AU-rich element with similar affinities. Finally, we demonstrate that the interaction of the RNase PH domain of PM-Scl-75 is readily competed by poly( U), but only inefficiently using other homopolymeric RNAs. These data demonstrate that RNase PH domains in general have an affinity for U- and AU-rich sequences, and broaden the potential role in RNA biology of proteins containing these domains.