Structural and biochemical characterisation of human SWI/SNF-related chromatin remodelling complexes

真核生物的基因组以染色质的形式存在。染色质的组织提供了一种调节潜在遗传信息的方法。真核生物调节染色质构象的一种方式是通过染色质重塑马达蛋白的作用。SWI/SNF重塑复合物的人类形式是由大约15个部分重叠的亚基组成的一系列染色质重塑ATP酶。这些复合物主要在增强子处起作用，并且已经观察到在一系列人类癌症中以高速率突变。这些复合物的活性的生物化学和结构表征是有限的。该提议旨在使用从昆虫细胞纯化的重组表达蛋白来解决这一问题。冷冻电子显微镜将被用来表征复合物的结构。将使用在模型DNA模板上组装的核小体进行生化测定。深入了解这些复合物的结构和功能将有助于识别癌细胞的脆弱性，其中增殖是由有缺陷的复合物驱动的。项目提供跨学科技能培训。

The genomes of eukaryotes exist as chromatin. The organisation of chromatin provides a means of regulating access to the underlying genetic information. One way in which eukaryotes regulate chromatin conformation is through the action of chromatin remodelling motor proteins. Human forms of the SWI/SNF remodelling complex are a series of chromatin remodelling ATPases comprised of some 15 partially overlapping subunits. These complexes function predominantly at enhancers and have been observed to be mutated at high rates in a range of human cancers. Biochemical and structural characterisation of the activities of these complexes is limited. This proposal aims to address this using recombinant expressed proteins purified from insect cells. Cryo-electron microscopy will be used to characterise the structure of the complexes. Biochemical assays will be performed using nucleosomes assembled on model DNA templates. Insight into the structure and function of these complexes will help identify vulnerabilities in cancer cells where proliferation is driven by defective complexes. Project provides training in Interdisciplinary Skills.