Measuring DNA Dynamics in Promoter Chromatin with Internally Labeled FRET-Capable Probes
Moran, Crystal R.1,2 and Woodbury, Neal2,3
1School of Life Sciences, Arizona State University; 2Center for BioOptical Nanotechnolgy, The Biodesign Institute at Arizona State University; 3Department of Chemistry and Biochemistry, Arizona State University
DNA condensation has been well-characterized to the point where we now know the dimensions and structure of the nucleosome, down to the precise number of DNA bases pairs that are wrapped around the histone core. It is well understood that this “beads-on-a-string” model is essential to package the approximately 3 billion base pairs of the human genome (nearly 2 m in length) into the confines of the cellular nucleus (d ≈ 5μm). However, we have yet to fully understand the dynamic events that allow cellular machinery (i.e. polymerases) to access the DNA usually sequestered in the nucleosomes. The goal of this project is to engineer a replica of the Mouse Mammary Tumor Virus (MMTV) promoter that will utilize Fluorescence Resonance Energy Transfer (FRET) to visualize DNA dynamics within promoter chromatin. We hypothesize that such experimentation will be critical in gaining insight into the mechanisms of hormone-dependent chromatin remodeling.