Understanding the Architecture, Assembly, and Interactome of the Nucleosome Remodelling and Deacetylase (NuRD) Complex

Abstract

This thesis presents data to enhance our understanding of the composition, assembly, and structure of the nucleosome remodeling and deacetylase (NuRD) complex, a transcriptional coregulator that is strongly linked to cancer. Understanding the structure and function of the gene regulating complexes will lead to a better understanding of mechanisms of gene regulation. This should ultimately both allow insight into disease processes that originate from gene regulatory network malfunction and might enable reprogramming of the gene expression for application in health, agriculture and industry. Chapter 1 represents a general introduction to gene regulation and the NuRD complex. The NuRD complex is a 1mDa multisubunit transcriptional coregulator protein complex, which is widely expressed and harbours both chromatin remodelling and histone deacetylase activities. Abnormalities in a number of its components are associated with various types of cancers and neurodegenerative diseases. Although the core components of the complex have been well established, the structural basis of the complex has poorly been studied. Chapter 2 describes the details of the materials and methods used in this research. Chapter 3 investigates NuRD complex stoichiometry and interactome using a label-free quantitative proteomics method and provides biochemical evidence for two new direct interactors of the complex. Chapter 4 and 5 investigate the recombinantly produced subcomplexes using biochemical and a mass spectrometry-based hybrid method for structural modelling of the NuRD complex. Chapter 6 discuss the findings and outlines the future directions.