Environmental Molecular Sciences Laboratory

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Toward the visualization of NHEJ DNA repair machineries by cryo-EM

Date: 
Friday, May 15, 2020
Principal Investigator: 
Yuan He
Lead Institution: 
Northwestern University
Project ID: 
51361
Abstract: 

Among all different types of DNA damages, double strand breaks (DSBs) are viewed as the most toxic ones that lead to genome instability. Non-homologous end-joining (NHEJ) pathway, in which the two broken DNA ends are directly ligated without referring to a homologous template, is the primary DSB repair pathway that remains active throughout the cell cycle. We were able to reconstitute in vitro the NHEJ repair complex in two different states and produce preliminary cryo-EM reconstructions at resolution 5-10 Å using a 200kev site-entry screening microscope. Determining the structure of these NHEJ repair complexes at higher resolution is an essential step to our understanding of the coupling between DSB sensing and the final ligation and will be of broad significance to our understanding of DSB repair regulation as well as other DNA damage response directed by PI3K-related kinases.