Huntington's Disease Protein Huntingtin Associates with its own mRNA.
Huntington's Disease Protein Huntingtin Associates with its own mRNA.
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DOI：
10.3233/jhd-150177
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Tanese N
中科院分区：
文献类型：
--
作者：
Culver BP;DeClercq J;Dolgalev I;Yu MS;Ma B;Heguy A;Tanese N
背景：亨廷顿病（HD）蛋白亨廷顿蛋白（Htt）在多种细胞途径中发挥作用。突变的Htt蛋白对这些途径中的一个或多个通路的解除管制已被认为有助于疾病的发病机制。我们最近基于发现的蛋白质组学研究发现了与从小鼠大脑中纯化的内源性Htt蛋白相关的RNA结合蛋白和翻译因子，这表明Htt在RNA转运和翻译中的潜在新作用。目的：研究Htt对RNA代谢的影响，纯化并分析与Htt相关的RNA。方法：从免疫纯化的含htt蛋白复合物中提取RNA，采用微阵列和RNA- seq分析。结果：令人惊讶的是，与Htt共纯化的最富集的mRNA是Htt mRNA本身。Htt蛋白和Htt mRNA的关联独立于完整的核糖体被检测到，这表明它不是一种正在进行翻译的RNA。此外，我们在含有突变Htt蛋白的免疫沉淀中发现了最近报道的错误剪接的Htt mRNA，该mRNA编码由外显子1和部分下游内含子组成的截断蛋白。我们发现Htt蛋白与Htt mRNA共定位，并且野生型Htt减少了包含Htt 3 ' UTR的报告结构的表达。结论：HD蛋白与其自身的mRNA和RNA结合蛋白及翻译因子形成复合物。Htt可能通过转录后通路调节其表达。Htt可能具有与其他神经退行性疾病相关的RNA结合蛋白（如TDP-43和FUS）相似的机制特性。
Background: The Huntington’s disease (HD) protein huntingtin (Htt) plays a role in multiple cellular pathways. Deregulation of one or more of these pathways by the mutant Htt protein has been suggested to contribute to the disease pathogenesis. Our recent discovery-based proteomics studies have uncovered RNA binding proteins and translation factors associated with the endogenous Htt protein purified from mouse brains, suggesting a potential new role for Htt in RNA transport and translation. Objective: To investigate how Htt might affect RNA metabolism we set out to purify and analyze RNA associated with Htt. Methods: RNA was extracted from immunopurified Htt-containing protein complexes and analyzed by microarrays and RNA-Seq. Results: Surprisingly, the most enriched mRNA that co-purified with Htt was Htt mRNA itself. The association of Htt protein and Htt mRNA was detected independent of intact ribosomes suggesting that it is not an RNA undergoing translation. Furthermore, we identified the recently reported mis-spliced Htt mRNA encoding a truncated protein comprised of exon 1 and a portion of the downstream intron in the immunoprecipitates containing mutant Htt protein. We show that Htt protein co-localizes with Htt mRNA and that wild-type Htt reduces expression of a reporter construct harboring the Htt 3’ UTR. Conclusions: HD protein is found in a complex with its own mRNA and RNA binding proteins and translation factors. Htt may be involved in modulating its expression through post-transcriptional pathways. It is possible that Htt shares mechanistic properties similar to RNA binding proteins such as TDP-43 and FUS implicated in other neurodegenerative diseases.