Unravelling key cellular machinery for spliceosome regulation

前体信使RNA（pre-mRNA）剪接是真核细胞中一个重要的和严格调控的过程。在转录过程中，它允许通过去除非编码区域并重新排列部分来揭示制造蛋白质的信息，作为使指令可读的方式，就像解密代码一样。通过这样做，mRNA剪接不仅提供了一种调节基因表达的方法，而且还有助于增加我们蛋白质组的多样性，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，在不同组织中表达或具有不同功能的蛋白质变体。因此，剪接在我们的健康中起着重要作用，但当它出错时可能导致疾病。例如，已经鉴定了> 15，000个癌症特异性错误剪接事件。 剪接正在成为治疗药物开发的一个有趣的靶点，这反映在许多直接靶向RNA的生物技术公司的出现，以及许多对这一领域感兴趣的制药公司。然而，用小分子靶向RNA已经被证明是非常具有挑战性的，需要对这个复杂系统有更多的了解，以促进未来医学的发展。剪接是由称为剪接体的多组分蛋白质复合物进行的。剪接体的一些组分需要翻译后催化裂解成较小的片段以掺入剪接体复合物中。然而，关于什么酶切割这些成分，这个过程是如何调节的，以及它在剪接中的作用，我们知之甚少。该项目旨在通过采用高度多学科的方法来揭示这种未探索和非常令人兴奋的调控机制，包括蛋白质工程，结构生物学，定量蛋白质组学和细胞生物学技术，以帮助在分子水平上识别和表征负责剪接组分蛋白水解加工的蛋白酶以及此类事件的下游效应。这可能揭示新的可药用靶点和间接调节剪接的替代方法，而不依赖于靶向RNA。

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.