Understanding how the NuRD complex regulates ES cell differentiation using single molecule fluorescence imaging

使用单分子荧光成像了解 NuRD 复合物如何调节 ES 细胞分化

• 批准号: MR/M010082/1
• 负责人: Ernest Laue
• 金额: $ 47.98万
• 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: 2014
• 资助国家: 英国
• 起止时间: 2014 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FM010082%2F1
• 关键词: Understanding; how; NuRD; complex; regulates

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 cell in the body.The project brings together a unique combination of skill sets, in biochemistry/structural biology, single molecule imaging, and stem cell biology, to develop single molecule approaches for studying nuclear processes in mammalian cells. Up to now most work in this field has focussed on studies of processes occurring on either the cell surface or in bacterial cells. We have carried out a great deal of preliminary work to demonstrate feasibility (and thus reduce the risk), such that we can now envisage a highly inter-disciplinary research program to study the way the NuRD complex interacts with chromatin, and how this controls gene expression during ES cell differentiation. 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. In a related project we are attempting to develop small molecule inhibitors and activators of NuRD complexes to control chromatin structure. Our research may thus 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)蛋白复合体在控制我们的基因组在细胞内包装成一种称为染色质的结构的方式中发挥着关键作用。这种包装反过来控制特定基因(DNA序列)是否表达/不表达。特别是，当胚胎干细胞(ES)在体内首次开始分化为所有不同类型的特化细胞时，NuRD复合体控制着基因的表达。该项目结合了生物化学/结构生物学、单分子成像和干细胞生物学的独特技能组合，开发了研究哺乳动物细胞核过程的单分子方法。到目前为止，这一领域的大部分工作都集中在细胞表面或细菌细胞内发生的过程的研究上。我们已经开展了大量的前期工作来证明可行性(从而降低风险)，这样我们现在就可以设想一个高度跨学科的研究计划，研究NuRD复合体与染色质相互作用的方式，以及这如何控制ES细胞分化过程中的基因表达。我们的长期目标是利用这一认识来控制干细胞的分化。当这种理解应用于胚胎干细胞或已被诱导成为干细胞(iPS细胞)的成年细胞时，可能具有巨大的潜力--例如，为研究疾病进展提供人体组织来源，或开发个性化分子疗法的药物。在一个相关的项目中，我们正在尝试开发NuRD复合体的小分子抑制剂和激活剂来控制染色质结构。因此，从长远来看，我们的研究可能会促进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