Sequence Determinants of In Vivo and Intrinsic Nucleosome Occupancy

The genomes of all eukaryotic organisms are packaged into chromatin, the fundamental unit of which is the nucleosome. Since the proposal of the nucleosome as the primary repeating unit of chromatin structure in 1974, it has become clear that the biological roles of nucleosomes extend far beyond simple DNA packaging and include virtually all processes involving the genome. Despite the integral roles of nucleosomes in many fundamental biological processes, the relative contributions of the cellular cues and sequence features that directly govern their arrangement on genomic DNA remain unclear. In this Thesis, I characterise the sequence preferences of nucleosomes using data sets derived from genome-wide studies. I describe the analysis of data derived from the first genome-wide map of in vivo nucleosome occupancy across a eukaryotic genome (in this case, the budding yeast, Saccharomyces cerevisiae). Using these data, I construct a sequence-based linear model of nucleosome occupancy that takes into account structural features of DNA (which correlate with simple base composition) as well as transcription factor (TF) binding site information, which has significant ability to predict nucleosome occupancy in vivo. I go on to test particular aspects of this model and show that genetic perturbation of TFs that the in vivo model deems important (Abf1, Reb1, and Rsc3) have the expected effects, an increase in nucleosome occupancy over their cognate binding sites as well as a reduction in transcription from the corresponding genes, suggesting that these factors are required for promoter function and definition. I also confirm that in vitro nucleosome occupancy correlates highly with sequence features important for nucleosome occupancy in vivo and go on to develop a simple model for nucleosome occupancy based solely on histone-DNA interactions. This model suggests that base composition (G+C content) is a dominant feature in determining intrinsic nucleosome occupancy. Finally, I apply a model of intrinsic nucleosome occupancy to the human genome and show that there is a fundamental difference in intrinsic nucleosome occupancy at regulatory regions across species. This finding illustrates a potential functional consequence of variation in base composition in eukaryotic genomes.Ph