NSF Award
2321501
Single particle Cryo-electron microscopy (Cryo-EM) is a powerful tool for studying the structure of proteins and other molecules in our cells. The ability to capture exquisitely detailed images of proteins, protein complexes, antibodies, and viruses has been facilitated by recent advancements in Cryo-EM technology, aptly termed the "resolution revolution." With the support of Cryo-EM experts at the University of Michigan and the National Center for Cryo-EM Access and Training (NCCAT) in New York, the PI will receive training and gain new expertise in using single-particle Cryo-EM methods to answer key scientific questions surrounding gene regulatory mechanisms. This training in single particle cryoEM will allow the PI to obtain protein-DNA structures at high resolution that are not currently available through X-ray crystallography or NMR due to size and other limitations. Cryo-EM has become an essential component of modern biological research, and there is a great need to train a highly skilled workforce to meet the growing demand. By becoming an expert in this area, the PI will revitalize an existing University of Vermont (UVM) Cryo-EM facility and set up training for other UVM faculty, lab personnel, postdocs, graduate, and undergraduate students.<br/><br/>The primary objective of this project is to acquire the ability to lead and conduct single particle Cryo-electron microscopy (Cryo-EM) experiments to elucidate the molecular structures of large proteins and oligomeric complexes. Specifically, this project will investigate how chromatin reader domains, such as the bromodomain, interact with post-translational modifications on histone proteins and the nucleosome to regulate essential DNA templated processes. Utilizing Cryo-EM to collect structural data on the native complexes formed within the cell will be essential to further current knowledge of how epigenetic signaling regulates critical cellular processes. To obtain this expertise, the PI has developed a training plan to learn single-particle Cryo-EM techniques from experts at the University of Michigan and at the National Center for Cryo-EM Access and Training (NCCAT). Through these partnerships data processing and training workflows will be established to determine the structures of chromatin regulatory complexes. This collaborative effort will also be utilized to develop new educational and training resources that will facilitate access to Cryo-EM technology at the local and national levels.<br/><br/>This project is jointly funded by the Molecular Biophysics Program of the Molecular and Cellular Biosciences Division in the Biological Sciences Directorate and the Established Program to Stimulate Competitive Research (EPSCoR).<br/><br/>This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.
