Cold snapshots of DNA repair: Cryo-EM structures of DNA-PKcs and NHEJ machinery

• Published in: Progress in biophysics and molecular biology Vol. 186; pp. 1 - 13
• Main Authors: Amin, Himani, Zahid, Sayma, Hall, Chloe, Chaplin, Amanda K.
• Format: Journal Article
• Language: English
• Published: England 01.01.2024
• Subjects: DNA-PK; NHEJ; DNA repair; Cryo-EM
• ISSN: 0079-6107; 1873-1732; 1873-1732
• DOI: 10.1016/j.pbiomolbio.2023.11.007

The proteins and protein assemblies involved in DNA repair have been the focus of a multitude of structural studies for the past few decades. Historically, the structures of these protein complexes have been resolved by X-ray crystallography. However, more recently with the advancements in cryo-electron microscopy (cryo-EM) ranging from optimising the methodology for sample preparation to the development of improved electron detectors, the focus has shifted from X-ray crystallography to cryo-EM. This methodological transition has allowed for the structural determination of larger, more complex protein assemblies involved in DNA repair pathways and has subsequently led to a deeper understanding of the mechanisms utilised by these fascinating molecular machines. Here, we review some of the key structural advancements that have been gained in the study of non-homologous end joining (NHEJ) by the use of cryo-EM, with a focus on assemblies composed of DNA-PKcs and Ku70/80 (Ku) and the various methodologies utilised to obtain these structures.