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Researchers used single-molecule imaging in Xenopus egg extract to study how DNA double-strand breaks with different chemical structures are held together during repair by non-homologous end joining (NHEJ). They found that core NHEJ proteins maintain stable connections between DNA ends regardless of their chemical composition, with comparable formation and stability of repair complexes across nine different DNA substrate types. The protein XLF was essential for forming close-range repair complexes on all substrates, and its structural features became particularly important when DNA ends required additional processing before repair.
Why it matters
This work reveals how cells maintain physical contact between broken DNA ends during the repair process, which is critical for preventing chromosome rearrangements and maintaining genome stability. Understanding these mechanisms could inform therapeutic approaches for cancer treatment and improving DNA repair efficiency in various clinical contexts.
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⚠️ Preprint – Noch nicht peer-reviewed
Dieser Artikel wurde noch nicht von unabhängigen Experten begutachtet. Die Ergebnisse sind vorläufig und sollten mit Vorsicht interpretiert werden.
DNA double strand breaks (DSBs) generated by genotoxic stress frequently contain chemically diverse termini that require end processing before ligation by non-homologous end joining (NHEJ). How end chemistry influences assembly and persistence of NHEJ synaptic complexes remains unclear. Using single-molecule imaging approaches in Xenopus egg extract, we measured formation and stability of the long-range (LRC) and short-range (SRC) complexes on nine DNA substrates spanning kinase-, polymerase- and nuclease-dependent processing requirements. LRC formation and lifetime were comparable across end types, and SRC formation and stability were largely insensitive to end chemistry. DNA hairpins, physiological intermediates of V(D)J recombination, were a notable exception that formed LRCs but inefficiently transitioned to SRCs. Depletion of cognate processing factors had modest effects on SRC formation and persistence, indicating that core NHEJ factors provide the principal structural support for synapsis across diverse termini. Although XLF was required for SRC formation on all substrates, reducing an XLF dimer from two intact tail/Ku-binding motifs to one selectively reduced productive SRC formation on ends requiring processing. Together, these results support a model in which partially redundant protein-protein interactions between core NHEJ factors maintain synapsis while end-specific processing reactions shape repair outcome.
Source: Core NHEJ Factors Maintain Synapsis Across Diverse DNA End Structures