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Understanding the role of U5 snRNP gene mutation in pre-messenger RNA splicing and craniofacial development

了解 U5 snRNP 基因突变在前信使 RNA 剪接和颅面发育中的作用

基本信息

批准号：
BB/N000358/1
负责人：
Raymond O'Keefe
金额：
$ 57.44万
依托单位：
University of Manchester
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2016
资助国家：
英国
起止时间：
2016 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FN000358%2F1
关键词：
Understanding role U5 snRNP gene

项目摘要

The DNA of a cell is copied into a pre-messenger RNA (pre-mRNA) that the cell uses as a template for protein production. Some of the information contained in DNA is not required for making proteins, therefore, unwanted information must be removed before a protein is made. This unwanted information is removed, or spliced, from pre-mRNA by a process similar to the editing of unwanted frames from a film. This splicing of the pre-mRNA is very important because it must occur accurately in order for functional proteins to be produced. Splicing at the wrong position could have disastrous effects on the final protein produced. Abnormal proteins generated due to mistakes in splicing could cause defects in the development of an organism or result in disease.The process of splicing is carried out by a large RNA/protein complex called the spliceosome. The spliceosome interacts with the pre-mRNA to identify and splice out the unwanted regions. At the core of the spliceosome is the U5 snRNP. The U5 snRNP contributes to the active site of the spliceosome and orients the pre-mRNA for accurate removal of the unwanted regions from the pre-mRNA which are called introns. Therefore, the U5 snRNP is essential for the function of the spliceosome. We have recently found that mutations in genes that make proteins of the U5 snRNP lead to the craniofacial disorders Burn-McKeown Syndrome (BMKS) and MandibuloFacial Dysostosis, Guion-Almeida type (MFDGA). This observation suggests that, in some situations, mutation in essential splicing factors may only influence a subset of pre-mRNAs. Because patients with these mutations only present with very specific craniofacial defects, it appears that these mutations in the U5 snRNP only influence the splicing of some pre-mRNAs required at a specific developmental stage. It is not clear how only certain pre-mRNAs are influenced by these U5 snRNP gene mutations, thus we propose to investigate this important question during this project. We will take advantage of the high similarity between the human, mouse and yeast U5 snRNP proteins to investigate the exact defects associated with these U5 snRNP gene mutations in the experimentally tractable yeast system and with mouse and human cell lines. To gain an understanding of how U5 snRNP gene mutations cause defects in craniofacial development, we will also explore splicing defects directly in cranial neural crest cells from mice but also develop mouse models of these disorders. We will find pre-mRNAs where the splicing process has occurred at incorrect positions, creating errors that cause the formation of abnormal proteins. We will search for links between these abnormal proteins and craniofacial development to gain an understanding of why craniofacial development is disrupted by the mutations in U5 snRNP genes. Because the process of splicing is critical for cellular survival, gaining a better understanding of spliceosome function through the investigation of mutants with splicing defects informs our understanding of fundamental biological processes. Additionally, this research project will provide essential information on the role of pre-mRNA splicing in development and aid in the understanding of how mutations in core splicing factors can cause disease.

细胞的DNA被复制到前信使RNA(前信使RNA)中，细胞将其用作蛋白质生产的模板。DNA中包含的一些信息对于制造蛋白质来说并不是必需的，因此，在制造蛋白质之前必须去除不需要的信息。这种不想要的信息通过类似于编辑电影中不想要的帧的过程从Pre-mRNA中移除或拼接。Pre-mRNA的这种剪接非常重要，因为它必须准确地发生，才能产生功能蛋白。在错误的位置剪接可能会对最终产生的蛋白质产生灾难性的影响。剪接错误产生的异常蛋白质可能会导致生物体的发育缺陷或导致疾病。剪接过程是由一个被称为剪接体的大型RNA/蛋白质复合体执行的。剪接体与前信使核糖核酸相互作用，识别并剪接出不想要的区域。剪接体的核心是U5SnRNP。U5SnRNP参与剪接体的活性部位，并定位Pre-mRNA，准确地从Pre-mRNA中移除不需要的区域，称为内含子。因此，U5SnRNP对于剪接体的功能是必不可少的。我们最近发现，使U5单链RNP蛋白的基因突变导致颅面部疾病烧伤-麦基翁综合征(BMKS)和下颌面部骨质疏松症，Guion-Almeida类型(MFDGA)。这一观察结果表明，在某些情况下，基本剪接因子的突变可能只影响前mRNAs的一部分。由于具有这些突变的患者只存在非常特殊的头面部缺陷，因此U5 SNRNP中的这些突变似乎只影响特定发育阶段所需的一些前-mRNAs的剪接。目前尚不清楚只有某些前mRNAs是如何受到这些U5SnRNP基因突变的影响的，因此我们建议在这个项目中研究这个重要的问题。我们将利用人、小鼠和酵母U5 SnRNP蛋白之间的高度相似性，在实验上易于处理的酵母系统以及小鼠和人类细胞系中调查与这些U5 SnRNP基因突变相关的确切缺陷。为了了解U5 SnRNP基因突变是如何导致颅面发育缺陷的，我们还将探索直接从小鼠的脑神经脊细胞中剪接缺陷，并建立这些疾病的小鼠模型。我们会发现前mRNAs的剪接过程发生在错误的位置，造成错误，导致异常蛋白质的形成。我们将寻找这些异常蛋白与颅面发育之间的联系，以了解为什么U5 SNRNP基因突变会扰乱颅面发育。由于剪接过程对细胞的生存至关重要，通过对存在剪接缺陷的突变体的研究来更好地了解剪接体的功能，有助于我们对基本生物学过程的理解。此外，这项研究项目将提供有关前mRNA剪接在发育中的作用的基本信息，并有助于理解核心剪接因子的突变如何导致疾病。