Interactions of the DNA ligase IV-XRCC4 complex with DNA ends and the DNA-dependent protein kinase

Ling Chen, Kelly Trujillo, Patrick Sung, Alan E. Tomkinson

Research output: Contribution to journalArticle

176 Citations (Scopus)

Abstract

The DNA-dependent protein kinase (DNA-PK), consisting of Ku and the DNA-PK catalytic subunit (DNA-PKcs), and the DNA ligase IV-XRCC4 complex function together in the repair of DNA double-strand breaks by non-homologous end joining. These protein complexes are also required for the completion of V(D)J recombination events in immune cells. Here we demonstrate that the DNA ligase IV-XRCC4 complex binds specifically to the ends of duplex DNA molecules and can act as a bridging factor, linking together duplex DNA molecules with complementary but non-ligatable ends. Although the DNA end-binding protein Ku inhibited DNA joining by DNA ligase IV-XRCC4, it did not prevent this complex from binding to DNA. Instead, DNA ligase IV-XRCC4 and Ku bound simultaneously to the ends of duplex DNA molecules. DNA ligase IV-XRCC4 and DNA-PKcs also formed complexes at the ends of DNA molecules, but DNA-PKcs did not inhibit ligation. Interestingly, DNA-PKcs stimulated intermolecular ligation by DNA ligase IV-XRCC4. In the presence of DNA-PK, the majority of the joining events catalyzed by DNA ligase IV-XRCC4 were intermolecular because Ku inhibited intramolecular ligation, but DNA-PKcs still stimulated intramolecular ligation. We suggest that DNA-PKcs-containing complexes formed at DNA ends enhance the association of DNA ends via protein-protein interactions, thereby stimulating intermolecular ligation.

Original languageEnglish (US)
Pages (from-to)26196-26205
Number of pages10
JournalJournal of Biological Chemistry
Volume275
Issue number34
DOIs
StatePublished - Aug 25 2000

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DNA-Activated Protein Kinase
DNA Ligases
DNA
Catalytic Domain
Ligation
Joining
Molecules
DNA Ligase ATP
Proteins
Catalytic DNA
V(D)J Recombination
Double-Stranded DNA Breaks
DNA-Binding Proteins
Carrier Proteins
Repair

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Interactions of the DNA ligase IV-XRCC4 complex with DNA ends and the DNA-dependent protein kinase. / Chen, Ling; Trujillo, Kelly; Sung, Patrick; Tomkinson, Alan E.

In: Journal of Biological Chemistry, Vol. 275, No. 34, 25.08.2000, p. 26196-26205.

Research output: Contribution to journalArticle

Chen, Ling ; Trujillo, Kelly ; Sung, Patrick ; Tomkinson, Alan E. / Interactions of the DNA ligase IV-XRCC4 complex with DNA ends and the DNA-dependent protein kinase. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 34. pp. 26196-26205.
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