Description
Nuclear architecture is increasingly recognised as an important regulator of DNA target search. We have developed fluorescence microscopy approaches based on fluorescence fluctuation spectroscopy to quantify how molecules move through the complex DNA networks of living cell nuclei. These methods provide the spatiotemporal resolution needed to map how genome organisation shapes nuclear transport and multi-protein complex formation. Using this approach, we discovered that dynamic rearrangements of DNA generate a chromatin network that facilitates transcription factor (TF) recruitment to target sites. We are now extending these measurements to determine how TFs use protein–protein interactions to enhance the efficiency of DNA target search. By extracting spatiotemporal correlation functions directly from live-cell microscopy time series, we can resolve molecular movement from local diffusion to long-range nuclear transport, while distinguishing changes associated with oligomerisation and complex formation using spectrally distinct fluorescent ligands. The approach is compatible with single-, dual- and triple-channel confocal laser scanning microscopy (CLSM) and enables quantitative analysis of how TF stoichiometry regulates DNA target search and gene expression. Together, these studies position genome organisation as a dynamic roadmap that guides DNA-binding proteins through the nucleus and helps maintain genome function
| I am the presenting author | No |
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| If you are not the presenting author, please give the presenting author's name: | Elizabeth Hinde |