Year: 2018
Type: article
Abstract: To investigate the fidelity of canonical non-homologous end joining (C-NHEJ), we developed an assay to detect EJ between distal ends of two Cas9-induced chromosomal breaks that are joined without causing insertion/deletion mutations (indels). Here we find that such EJ requires several core C-NHEJ factors, including XLF. Using variants of this assay, we find that C-NHEJ is required for EJ events that use 1-2, but not ≥3, nucleotides of terminal microhomology. We also investigated XLF residues required for EJ without indels, finding that one of two binding domains is essential (L115 or C-terminal lysines that bind XRCC4 and KU/DNA, respectively), and that disruption of one of these domains sensitizes XLF to mutations that affect its dimer interface, which we examined with molecular dynamic simulations. Thus, C-NHEJ, including synergistic function of distinct XLF domains, is required for EJ of chromosomal breaks without indels.
Source: Nature Communications
Institution: Beckman Research Institute
Language: English
Topic: DNA Repair Mechanisms
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $6,990
Awards: G1259816303, Correct end use during end joining and radioresistance, G2559336095, G2598592983, G2962418797, G344711537, Computationally Guided Design of Thermostable mutants of GPCR-transducer complexes, G453899893, G5077536465, Regulation of Single Strand Annealing Repair of Mammalian Chromosomal Breaks, Genomic and Microenvironment Analysis of HIV-Associated Diffuse Large B-cell Lymphoma (Immuno/Microenvironment), G7614147705, G772794905, G9000023467, G9000138537, G9000193878, G9000344495
OpenAlex ID: W2809529383