Understanding how the NuRD complex assembles and functions in mouse embryonic stem cells (mESC's)

了解 NuRD 复合物如何在小鼠胚胎干细胞 (mESC) 中组装和发挥作用

• 批准号: MR/P019471/1
• 负责人: Ernest Laue
• 金额: $ 271.68万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2017
• 资助国家: 英国
• 起止时间: 2017 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FP019471%2F1
• 关键词: Understanding; how; NuRD; complex; assembles

The Nucleosome Remodeling and Deacetylation (NuRD) protein complex plays a key role in controlling the way our genomes are packaged inside the cell into a structure called chromatin. This packaging in turn controls whether particular genes (sequences of DNA) are/are not expressed. In particular, the NuRD complex controls gene expression as embryonic stem (ES) cells first start to differentiate into all the different types of specialised cells in the body. Without some of the NuRD complex components, ES cells cannot differentiate at all, clearly demonstrating its importance. So how does the NuRD complex form? And how does it affect gene expression in ES cells? These are the questions we hope to answer.Up until now, we (and the field as a whole) have focussed on trying to understand which components make up the NuRD complex, which regions of the genome it interacts with, and which genes are affected by it. However, we have recently shown that we can make the individual components and assemble the NuRD complex outside of a cell. This will allow us to study its structure and how it interacts with the small regions of the genome to which it binds. We have shown using cutting-edge imaging that we can track single NuRD complex components inside a cell and watch in real-time how they assemble on chromatin. We can also study how the NuRD complex affects the binding of other proteins, and ultimately gene expression. We now envisage a highly inter-disciplinary research program that combines these approaches to determine the structure of the NuRD complex, understand how NuRD complexes assemble and interact with different parts of the genome, and how they control gene expression. Our long-term goal is to use this understanding to control the differentiation of stem cells. This understanding when applied to either ES cells, or adult cells that have been induced to become stem cells (iPS cells), could have enormous potential - e.g. for providing a source of human tissue to study disease progression, or to develop drugs for personalised molecular therapies. We will also attempt to develop small molecule inhibitors and activators of NuRD complexes to control chromatin structure. Our research may in the long-term facilitate our ability to directly influence gene expression profiles, stem cell differentiation and disease.

核小体重塑和去乙酰化（NuRD）蛋白复合物在控制我们的基因组在细胞内包装成染色质结构的方式方面起着关键作用。这种包装反过来控制特定基因（DNA序列）是否表达。特别是，当胚胎干细胞（ES）首先开始分化成体内所有不同类型的特化细胞时，NuRD复合体控制基因表达。没有一些NuRD复杂成分，胚胎干细胞根本无法分化，这清楚地表明了它的重要性。那么NuRD复合体是如何形成的呢？它是如何影响胚胎干细胞的基因表达的？这些都是我们希望回答的问题。到目前为止，我们（以及整个领域）一直专注于试图了解NuRD复合体的组成成分，它与基因组的哪些区域相互作用，以及哪些基因受其影响。然而，我们最近已经证明，我们可以在细胞外制造单独的组件并组装NuRD复合体。这将使我们能够研究它的结构以及它如何与它所结合的基因组的小区域相互作用。我们已经展示了使用尖端成像技术，我们可以跟踪细胞内单个NuRD复杂组分，并实时观察它们如何在染色质上组装。我们还可以研究NuRD复合物如何影响其他蛋白质的结合，并最终影响基因表达。我们现在设想一个高度跨学科的研究计划，结合这些方法来确定NuRD复合体的结构，了解NuRD复合体如何组装并与基因组的不同部分相互作用，以及它们如何控制基因表达。我们的长期目标是利用这种认识来控制干细胞的分化。当将这种理解应用于胚胎干细胞或被诱导成为干细胞（iPS细胞）的成人细胞时，可能具有巨大的潜力——例如，为研究疾病进展提供人体组织的来源，或开发用于个性化分子治疗的药物。我们还将尝试开发NuRD复合物的小分子抑制剂和激活剂来控制染色质结构。从长远来看，我们的研究可能有助于我们直接影响基因表达谱、干细胞分化和疾病的能力。

项目成果

FRET-enhanced photostability allows improved single-molecule tracking of proteins and protein complexes in live mammalian cells.

• DOI: 10.1038/s41467-018-04486-0
• 发表时间: 2018-06-28
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Basu S;Needham LM;Lando D;Taylor EJR;Wohlfahrt KJ;Shah D;Boucher W;Tan YL;Bates LE;Tkachenko O;Cramard J;Lagerholm BC;Eggeling C;Hendrich B;Klenerman D;Lee SF;Laue ED
• 通讯作者: Laue ED

Three-Dimensional Super-Resolution in Eukaryotic Cells Using the Double-Helix Point Spread Function.

• DOI: 10.1016/j.bpj.2017.02.023
• 发表时间: 2017-04-11
• 期刊: Biophysical journal
• 影响因子: 3.4
• 作者: Carr AR;Ponjavic A;Basu S;McColl J;Santos AM;Davis S;Laue ED;Klenerman D;Lee SF
• 通讯作者: Lee SF

Three-Dimensional Super-Resolution in Eukaryotic Cells Using the Double-Helix Point Spread Function

使用双螺旋点扩散函数的真核细胞三维超分辨率

• DOI: 10.17863/cam.9529
• 发表时间: 2017
• 作者: Carr A

Combining fluorescence imaging with Hi-C to study 3D genome architecture of the same single cell.

将荧光成像与 Hi-C 相结合，研究同一单细胞的 3D 基因组结构。

• DOI: 10.1038/nprot.2018.017
• 发表时间: 2018
• 期刊: Nature protocols
• 影响因子: 14.8
• 作者: Lando D

Publisher Correction: Live-cell three-dimensional single-molecule tracking reveals modulation of enhancer dynamics by NuRD.

出版商更正：活细胞三维单分子追踪揭示了 NuRD 对增强子动力学的调节。

• DOI: 10.1038/s41594-023-01179-1
• 发表时间: 2024
• 期刊: Nature structural & molecular biology
• 影响因子: 16.8
• 作者: Basu S