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The essential elements for the noncovalent association of two DNA ends during NHEJ synapsis
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Journal Article
The essential elements for the noncovalent association of two DNA ends during NHEJ synapsis
2019
Overview
One of the most central questions about the repair of a double-strand DNA break (DSB) concerns how the two free DNA ends are brought together — a step called synapsis. Using single-molecule FRET (smFRET), we show here that both Ku plus XRCC4:DNA ligase IV are necessary and sufficient to achieve a flexible synapsis of blunt DNA ends, whereas either alone is not. Addition of XLF causes a transition to a close synaptic state, and maximum efficiency of close synapsis is achieved within 20 min. The promotion of close synapsis by XLF indicates a role that is independent of a filament structure, with action focused at the very ends of each duplex. DNA-PKcs is not required for the formation of either the flexible or close synaptic states. This model explains in biochemical terms the evolutionarily central synaptic role of Ku, X4L4, and XLF in NHEJ for all eukaryotes.
During a process termed synapsis, the two DNA ends at a double-strand break (DSB) are brought together into physical proximity. Here, the authors use a single-molecule FRET approach with purified proteins to investigate the mechanism of synapsis in DSB repair by non-homologous DNA end joining (NHEJ).
Publisher
Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
Subject
/ 82/83
/ DNA
/ DNA Repair Enzymes - genetics
/ DNA Repair Enzymes - metabolism
/ DNA-Binding Proteins - genetics
/ DNA-Binding Proteins - metabolism
/ DNA-dependent protein kinase
/ Fluorescence Resonance Energy Transfer
/ Humanities and Social Sciences
/ Recombinant Proteins - genetics
/ Recombinant Proteins - metabolism
/ Science
