Organization and Dynamics of the Nonhomologous End-joining Machinery During DNA Double-strand Break Repair

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2015 May 6

PMID 25941401

Citations 96

Authors

Dylan A Reid

Sarah Keegan

Alejandra Leo-Macias

Go Watanabe

Natasha T Strande

Howard H Chang

Betul Akgol Oksuz

David Fenyo

Michael R Lieber

Dale A Ramsden

Eli Rothenberg

Affiliations

Soon will be listed here.

Abstract

Nonhomologous end-joining (NHEJ) is a major repair pathway for DNA double-strand breaks (DSBs), involving synapsis and ligation of the broken strands. We describe the use of in vivo and in vitro single-molecule methods to define the organization and interaction of NHEJ repair proteins at DSB ends. Super-resolution fluorescence microscopy allowed the precise visualization of XRCC4, XLF, and DNA ligase IV filaments adjacent to DSBs, which bridge the broken chromosome and direct rejoining. We show, by single-molecule FRET analysis of the Ku/XRCC4/XLF/DNA ligase IV NHEJ ligation complex, that end-to-end synapsis involves a dynamic positioning of the two ends relative to one another. Our observations form the basis of a new model for NHEJ that describes the mechanism whereby filament-forming proteins bridge DNA DSBs in vivo. In this scheme, the filaments at either end of the DSB interact dynamically to achieve optimal configuration and end-to-end positioning and ligation.

Citing Articles

Single-molecule techniques in studying the molecular mechanisms of DNA synapsis in non-homologous end-joining repair.

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Multivalent interactions of the disordered regions of XLF and XRCC4 foster robust cellular NHEJ and drive the formation of ligation-boosting condensates in vitro.

Vu D, Bonucci A, Breniere M, Cisneros-Aguirre M, Pelupessy P, Wang Z Nat Struct Mol Biol. 2024; 31(11):1732-1744.

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