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RNA-binding landscape of Rrm4, a key RNA-binding protein mediating mRNA transport in Ustilago maydis

Rrm4 的 RNA 结合景观，Rrm4 是玉米黑粉菌中介导 mRNA 运输的关键 RNA 结合蛋白

基本信息

批准号：
242463178
负责人：
Professor Dr. Michael Feldbrügge
金额：
--
依托单位：
Institut für Mikrobiologie
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2014
资助国家：
德国
起止时间：
2013-12-31 至 2017-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/242463178?language=en
关键词：
RNA binding landscape Rrm4 key

项目摘要

RNA-binding proteins are key factors of posttranscriptional gene regulation. Commonly, they contain more than one RNA-binding domain for interaction with target mRNAs. A key process mediated by RNA-binding proteins is long-distance transport of mRNAs along microtubules. This is particularly important in highly polarized cells such as mammalian neurons or fungal hyphae. An important approach to uncover the precise function of these RNA-binding proteins is the identification of target mRNAs and binding sites at the transcriptome-wide level. Recently, we showed that the RNA-binding protein Rrm4 containing three RNA recognition motifs is the major factor regulating microtubule-dependent mRNA transport in infectious hyphae of the plant pathogenic fungus Ustilago maydis. Here, we apply an improved iCLIP approach, in vivo UV crosslinking and immune precipitation, to identify target mRNAs and binding sites transcriptome-wide and at single nucleotide resolution. Key findings will be verified under non-native conditions using heterologous systems or in vitro approaches. Furthermore, we will address whether Rrm4 carries out global functions such as regulating mRNA processing, stability or translation. The obtained results will provide new insights into the interaction of eukaryotic RNA-binding proteins regulating mRNA transport.

RNA结合蛋白是转录后基因调控的关键因子。通常，它们含有一个以上的RNA结合结构域，用于与靶mRNA相互作用。由RNA结合蛋白介导的一个关键过程是mRNA沿沿着微管的长距离运输。这在高度极化的细胞中特别重要，例如哺乳动物神经元或真菌菌丝。揭示这些RNA结合蛋白的精确功能的一个重要方法是在转录组水平上鉴定靶mRNA和结合位点。最近，我们发现，RNA结合蛋白Rrm4含有三个RNA识别基序是调控微管依赖的mRNA运输的植物病原真菌玉米黑粉菌的感染性菌丝的主要因素。在这里，我们应用一种改进的iCLIP方法，体内UV交联和免疫沉淀，以确定目标mRNA和结合位点的转录组范围和单核苷酸分辨率。关键发现将在非天然条件下使用异源系统或体外方法进行验证。此外，我们将讨论Rrm4是否执行全局功能，如调节mRNA加工，稳定性或翻译。所获得的结果将提供新的见解的相互作用的真核RNA结合蛋白调节mRNA的运输。