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

点击翻译按钮获取中文摘要

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.