Unravelling key cellular machinery for spliceosome regulation

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Precursor-messenger RNA (pre-mRNA) splicing is an essential and tightly regulated process in eukaryotic cells. It allows, during the transcription process, to reveal the information to make proteins, by removing the not coding regions and rearranging sections, as way to make the instructions readable, much like decrypting a code. By doing so, mRNA splicing does not only provide a way to regulate gene expression, but also contributes to increase the diversity of our proteome by creating different variation for the same gene to give rise to different proteins, for example, expressed in different tissues or protein variants with different function. Therefore, splicing plays fundamental role in our health but can lead to disease when it goes awry. For example, >15,000 of cancer-specific mis-splicing events have been identified. Splicing is becoming an interesting target for the development of therapeutics, reflected by the emergence of many biotech companies targeting RNA directly, and many pharmaceutical companies interested in this area. However, targeting RNA with small molecules has been proving very challenging and more understanding about this complex system is needed to enable the development of future medicine. Splicing is carried out by a multicomponent protein complex called the spliceosome. Some components of the spliceosome need to be post-translational catalytic cleavage in smaller fragments to be incorporated in the spliceosome complex. However, not much is known about what enzymes cleave these components, how this process is regulated, and its role in splicing. This project aims to shine light on this unexplored and very exciting regulatory mechanism by employing a highly multidisciplinary approach, with techniques including protein engineering, structural biology, quantitative proteomics, and cell biology to help to identify and characterize at the molecular level the protease(s) responsible for proteolytic processing of splicing components and the downstream effects of such events. This could reveal novel druggable targets and an alternative approach to modulate splicing indirectly, independent from targeting the RNA.