Investigating the difference between archaeal and eukaryal nucleosomal exchange using fluorescence resonance energy transfer

Brittany L. Pitrone, The College of Wooster


Since Archaea contain simplified forms of histones, studying archaeal histone interactions may provide a better understanding of how the more complex eukaryotic systems work to organize chromosomes. Provided that Archaea and Eukarya contain similar structural and functional relationships involving histones and nucleosomes, we wanted to investigate how the archaeal nucleosomes differ from eukaryal nucleosomes in DNA binding and stability. To do this we attempted to reconstitute archaeal nucleosomes and will use fluorescence resonance energy transfer (FRET) to determine how archaeal histones stably associate as a histone complex as well as with a given piece of DNA. In order to achieve our research objectives, the protein and DNA used in these experiments were purified, concentrated, and analyzed. A single attempt to reconstitute archaeal nucleosomes was unsuccessful; therefore further experiments (nucleosome reconstitution and FRET) must be conducted in order to address our research objectives. Overall, the results of the experiments will determine the significance of the archaeal nucleosomes optimization for increased flexibility rather than for stability, and how this affects archaeal histones binding affinity for DNA and for each other. Regardless of whether we determined how the archaeal nucleosomes optimization for flexibility rather than for stability affects archaeal histone binding affinity for each other and for DNA, a significant step towards comparing archaeal and eukaryal nucleosomal exchange was achieved.


© Copyright 2009 Brittany L. Pitrone