This subproject is one of many research subprojects utilizing theresources provided by a Center grant funded by NIH/NCRR. The subproject andinvestigator (PI) may have received primary funding from another NIH source,and thus could be represented in other CRISP entries. The institution listed isfor the Center, which is not necessarily the institution for the investigator.The proposed experiments will be a continuation of fluorescence dynamics experiments we have done with Dr. Digman at the LFD. This project examines the intracellular dynamics of DNA polymerases and associated signaling proteins in response to DNA damage. These proteins are required for DNA damage-induced mutagenesis, but the cell and molecular biology of such lesion bypass is very poorly understood. The application of advanced fluorescence dynamics techniques in living human cells is particularly appropriate for these studies since higher-order nuclear structure is almost certainly required for DNA polymerase switching events in response to blocked replication forks. In the proposed experiments we will investigate functional interactions using FRET/FLIM techniques. The appropriate expression vectors have been constructed. We propose to come to the LFD 9/17-21/07. WGM has been in touch with MD, and these dates are OK with her.
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