Final Activity Report Summary - ADAM WEST U.GLA (The role of chromatin insulator elements in the control gene expression)
Chromatin insulator elements allow genes to maintain programmes of expression independent of their chromosomal environment. My group studied the mechanism of insulators and their functions in the genome. Our work was largely based on previous studies of the paradigm insulator element called HS4. We identified a number of HS4-binding proteins and one of these proteins, called beta globin protein1 (BGP1), acted as a barrier to the encroachment of chromosomal gene silencing. Little was known about how insulator elements could act as barriers to silencing in vertebrates.
The first main objective of this project was to identify BGP1-interacting proteins. This would allow us to determine which activities were recruited by the barrier protein BGP1. Another project in our group studied deoxyribonucleic acid (DNA) methylation profiles at the HS4 insulator. This work provided evidence for a role of BGP1 in preventing DNA methylation. The identification of BGP1 co-factors would be a big step forward in understanding how BGP1 regulated DNA methylation, and may had broad implications in understanding how DNA methylation patterns were controlled in general. We developed an efficient procedure that resulted in the purification of a BGP1 protein complex, and, by the time of the project completion, we were identifying the proteins in this complex.
The second main objective of this work was to identify novel human insulator elements. Very few barrier elements were identified in vertebrates. This was because they were typically located in gene-poor areas that were rarely studied by researchers. It was of fundamental importance to understand how chromosomes were partitioned into functional units. It was not known whether barrier elements were primarily responsible, or how many barrier elements there might actually be. We therefore developed a strategy to efficiently isolate fragments of genomic DNA that were bound by BGP1 in cells. By the time of reporting we were identifying these sequences using DNA microarray analysis.
The first main objective of this project was to identify BGP1-interacting proteins. This would allow us to determine which activities were recruited by the barrier protein BGP1. Another project in our group studied deoxyribonucleic acid (DNA) methylation profiles at the HS4 insulator. This work provided evidence for a role of BGP1 in preventing DNA methylation. The identification of BGP1 co-factors would be a big step forward in understanding how BGP1 regulated DNA methylation, and may had broad implications in understanding how DNA methylation patterns were controlled in general. We developed an efficient procedure that resulted in the purification of a BGP1 protein complex, and, by the time of the project completion, we were identifying the proteins in this complex.
The second main objective of this work was to identify novel human insulator elements. Very few barrier elements were identified in vertebrates. This was because they were typically located in gene-poor areas that were rarely studied by researchers. It was of fundamental importance to understand how chromosomes were partitioned into functional units. It was not known whether barrier elements were primarily responsible, or how many barrier elements there might actually be. We therefore developed a strategy to efficiently isolate fragments of genomic DNA that were bound by BGP1 in cells. By the time of reporting we were identifying these sequences using DNA microarray analysis.