ADAM WEST U.GLA · The role of chromatin insulator elements in the control gene expression
FP6 — Marie Curie Actions (Human Resources and Mobility)
- Duration
- 2005-01-01 → 2006-12-31
- EU contribution
- €80,000
- Participants
- 1
- Scheme
- IRG
Lines connect the coordinator with its partners.
Results in brief
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.
Data: CORDIS, © European Union
Project objective
Chromatin insulators allow genes to maintain independent programmes of expression through the establishment of fixed chromatin boundaries. Vertebrate insulators have been found to regulate the expression of genes with functions from growth control and development to X-inactivation. The properties of insulators have been extensively characterised, but their mechanism of action is largely undetermined. The HS4 element from the chicken b-globin locus is the paradigm vertebrate insulator. Flanking transgenes with HS4 has proved useful in the generation of many transgenic animal and cell lines, by allowing uniform transgene expression in all tissue types.The aim of this proposal is to determine the mechanism(s) of insulator action. In my postdoctoral studies at the NIH (USA), I identified the first known insulator-binding proteins from vertebrates. I have recently taken up a lectureship at the University of Glasgow where I am establishing an independent research group to study chromatin insulator elements and gene regulation. I have minimal start-up funding, so I am applying for an IRG to help establish my research programme, thus promoting my re-integration into the European Research Area.The proposed work will focus on a new protein, called BGP1 that interacts with several binding sites within the HS4 element. BGP1 binding sites are required for the barrier activity of HS4. BGP1 binding sites are also required for the prevention of CpG DNA methylation, an epigenetic determinant of stable gene silencing. This is of broad interest as the promoters of tumour suppressor genes become aberrantly methylated and transcriptionally silenced in cancer. It is conceivable that BGP1 recruits a factor that mediates immunity from DNA methylation. I will purify BGP1-interacting proteins in order to identify factors that mediate BGP1-directed functions. I will also use BGP1 as bait to identify novel insulator elements to compare with the HS4 element.
Original text from CORDIS.
Participants
- UNIVERSITY OF GLASGOW · GLASGOWCoordinatorUnited Kingdom
Links
Data: CORDIS, © European Union