Dis3- and exosome subunit-responsive 3' mRNA instability elements.
Dis3- and exosome subunit-responsive 3' mRNA instability elements.
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Dis3 和外泌体亚基响应 3 mRNA 不稳定元件。
DOI：
10.1016/j.bbrc.2012.05.141
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发表时间：
2012
影响因子：
3.1
通讯作者：
Andrulis,ErikD
中科院分区：
文献类型：
--
作者：
Kiss,DanielL;Hou,Dezhi;Gross,RobertH;Andrulis,ErikD
真核生物的RNA周转在一定程度上受到外体的调节，外体是核糖核酸酶(RNase)和RNA结合蛋白的核质复合体。该复合体的主要核糖核酸酶被认为是Dis3，一种多功能的3‘-5’外切核糖核酸酶和内切核酸酶。虽然已知Dis3和核心外切体亚基被招募到转录活性基因和信使RNA(MRNA)底物上，但这种招募被认为是间接发生的。我们试图发现招募Dis3或其他外切体亚单位的顺式作用元件。使用一种名为RNA Scope的生物信息学工具从我们发表的Dis3耗竭来源的转录数据集中筛选上调转录的3‘非翻译区，我们确定了几个基序作为候选不稳定元件。使用荧光素酶报告系统进行的二次筛选显示，一个含有四种元素的盒式磁带破坏了报告记录的稳定性。基于RNAi的Dis3、RRP6、Rrp4、Rrp40或Rrp46的缺失降低了盒介导的不稳定的效果。对盒式磁带的截断分析表明，两个外切体亚单位敏感元件(ESSES)破坏了记者的稳定性。ESSE的点突变消除了这种失稳效应。对基于外切体亚单位缺失的微阵列的转录数据的检查表明，带有Esse的mRNAs存在于每个上调的mRNA数据集中，但在下调的数据集中表达不足或缺失。综上所述，我们的发现暗示了一种潜在的新的mRNA周转机制，涉及直接将Dis3和其他外体亚单位募集到和/或调控到mRNA底物上。
Eukaryotic RNA turnover is regulated in part by the exosome, a nuclear and cytoplasmic complex of ribonucleases (RNases) and RNA-binding proteins. The major RNase of the complex is thought to be Dis3, a multi-functional 3′–5′ exoribonuclease and endoribonuclease. Although it is known that Dis3 and core exosome subunits are recruited to transcriptionally active genes and to messenger RNA (mRNA) substrates, this recruitment is thought to occur indirectly. We sought to discover cis-acting elements that recruit Dis3 or other exosome subunits. Using a bioinformatic tool called RNA SCOPE to screen the 3′ untranslated regions of up-regulated transcripts from our published Dis3 depletion-derived transcriptomic data set, we identified several motifs as candidate instability elements. Secondary screening using a luciferase reporter system revealed that one cassette—harboring four elements—destabilized the reporter transcript. RNAi-based depletion of Dis3, Rrp6, Rrp4, Rrp40, or Rrp46 diminished the efficacy of cassette-mediated destabilization. Truncation analysis of the cassette showed that two exosome subunit-sensitive elements (ESSEs) destabilized the reporter. Point-directed mutagenesis of ESSE abrogated the destabilization effect. An examination of the transcriptomic data from exosome subunit depletion-based microarrays revealed that mRNAs with ESSEs are found in every up-regulated mRNA data set but are underrepresented or missing from the down-regulated data sets. Taken together, our findings imply a potentially novel mechanism of mRNA turnover that involves direct Dis3 and other exosome subunit recruitment to and/or regulation on mRNA substrates.
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影响因子：
3.3
作者：
Graham, AC;Kiss, DL;Andrulis, ED
通讯作者：
Andrulis, ED
影响因子：
4.5
作者：
Kiss, Daniel L.;Andrulis, Erik D.
通讯作者：
Andrulis, Erik D.
影响因子：
6.8
作者：
Arcus, V
通讯作者：
Arcus, V
外泌体招募 RNA 的机制
影响因子：
4.1
作者：
H. Malet;E. Lorentzen
通讯作者：
E. Lorentzen
DOI：
10.1073/pnas.0604731103
发表时间：
2006-08-08
影响因子：
11.1
作者：
Midtgaard, Soren F.;Assenholt, Jannie;Brodersen, Ditlev E.
通讯作者：
Brodersen, Ditlev E.