Unravelling key cellular machinery for spliceosome regulation

前体信使RNA(Pre-mRNA)剪接是真核细胞中一个必不可少且受到严格调控的过程。在转录过程中，它允许通过移除非编码区和重新排列部分来揭示制造蛋白质的信息，从而使指令变得可读，就像解密代码一样。通过这样做，mRNA剪接不仅提供了一种调控基因表达的方法，而且通过为同一基因创造不同的变异来产生不同的蛋白质，例如，在不同的组织中表达或功能不同的蛋白质变体，从而增加了我们蛋白质组的多样性。因此，剪接对我们的健康起着至关重要的作用，但当剪接出错时，可能会导致疾病。例如，已经确定了15,000个癌症特异性错误剪接事件。剪接正在成为治疗学发展的一个有趣的目标，反映在许多直接针对RNA的生物技术公司的出现和许多制药公司对这一领域的兴趣。然而，事实证明，以小分子靶向RNA是非常具有挑战性的，需要对这一复杂系统有更多的了解，才能使未来的医学发展成为可能。剪接是由称为剪接体的多组分蛋白质复合体进行的。剪接体的一些成分需要翻译后催化裂解成较小的片段，才能并入剪接体复合体。然而，关于是什么酶裂解这些成分，这一过程是如何调控的，以及它在剪接中的作用，人们知道的还不多。该项目旨在通过采用高度多学科的方法，利用蛋白质工程、结构生物学、定量蛋白质组学和细胞生物学等技术，在分子水平上帮助识别和表征负责剪接成分蛋白质分解处理的蛋白酶(S)以及此类事件的下游影响，从而揭示这一未知且非常令人兴奋的调控机制。这可能揭示新的可药物靶点和一种不依赖于靶向RNA的间接调节剪接的替代方法。

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.