Book of Abstracts: Albany 2011
June 14-18 2011
©Adenine Press (2010)
Mapping Oxidative Damage at High Resolution Throughout an Entire Genome
Oxidative damage to DNA has been proposed to be a major contributing factor to the process of ageing. Accumulation of oxidative damage has been associated with many degenerative diseases. Determining the locations of vulnerability to oxidative damage on a genome-wide scale will be pivotal to understanding the ageing process. One type of oxidative damage is produced by Reactive Oxygen species (ROS), primarily in the form of hydroxyl radicals, which abstract deoxyribose hydrogens along the DNA backbone, resulting in strand breaks. Regions of chromatin that encode essential genes that are highly expressed often are found in areas of relatively low chromatin condensation, and so may be more susceptible to ROS attack. These genomic regions are bound by numerous proteins, including transcription factors, and their accessibility is governed by nucleosome positioning.
We are developing a new method to biochemically process oxidatively damaged genomic DNA to make it suitable for high-throughput sequencing, in order to map oxidative damage throughout a genome at single-nucleotide resolution. Our initial experiments on a well-studied model system (1) demonstrate that this new method successfully identifies sites of oxidative damage. We will use these quantitative maps to determine which regions of a genome are particularly susceptible to oxidative damage, and which are resistant, and relate these damage maps to the underlying genes and functional regions of the genome. These studies will provide a view at unprecedented resolution of the spectrum of oxidative damage in the genome, to facilitate a deeper understanding of the relationship of DNA damage to ageing.
1Department of Chemistry
Boston, MA 02215