Skip to main content

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


EMSL 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.

Project Details

Start Date
2020-05-15
End Date
2021-03-17
Status
Closed

Team

Principal Investigator

Yuan He
Institution
Northwestern University

Team Members

Siyu Chen
Institution
Northwestern University

Janette Myers
Institution
Oregon Health & Science University

Irina El Khoury
Institution
Environmental Molecular Sciences Laboratory

Related Publications

Siyu Chen, Susan Fishbain, Yuan He, Linda Lee, Susan P. Lees-Miller, Tasmin Naila, Alan E. Tomkinson, Annie Wang. 2021. "Structural basis of long-range to short-range synaptic transition in NHEJ." Nature 593 (7858):294-298. https://doi.org/10.1038/s41586-021-03458-7