Cryo-Electron Microscopy of Histone Deacetylase Complexes

Class-1 histone deacetylases (HDACs 1, 2, 3) are essential enzymes present in the nucleus of all mammalian cells. They help regulate chromatin structure as the catalytic component of large multi-protein co-repressor complexes such as CoREST and NCOR/SMRT. The HDAC complexes play a role in many cellular processes, such as cancer, cell cycle progression and DNA repair. Investigating how these HDAC complexes act to control gene expression and their interactions with other proteins should lead to an understanding of their influence in the cell. Additionally, HDAC inhibitors are currently used to treat some cancers. An understanding of the structure and function of the complexes should mean that inhibitors can be designed to specific complexes to reduce side effects. There has recently been a revolution in Cryo-Electron Microscopy, which means that large protein and protein complex structures that previously could not be crystallised can now be solved at atomic resolution.

The aim of this PhD project is to solve the structure of the histone deacetylase complexes CoREST and NCOR/SMRT using Cryo-electron microscopy. Building upon previous work of the Schwabe group, we aim to obtain higher resolution structures, allowing for mechanistic insight into the complexes. By studying both complexes it will allow for comparison between their structure, function, and arrangement. This project will achieve this by producing the complexes the inclusion of their substrate, chromatin, as well as nanobodies to will help stabilise the complex. Additionally, we will incorporate emerging computational technologies to resolve flexible regions within the Cryo-EM data set.