The role of DNA dependent protein kinase in synapsis of DNA ends

Eric - Weterings, Nicole S. Verkaik, Hennie T. Brüggenwirth, Jan H J Hoeijmakers, Dik C. van Gent

Research output: Contribution to journalArticle

86 Citations (Scopus)

Abstract

DNA dependent protein kinase (DNA-PK) plays a central role in the non-homologous end-joining pathway of DNA double strand break repair. Its catalytic subunit (DNA-PKCS) functions as a serine/threonine protein kinase. We show that DNA-PK forms a stable complex at DNA termini that blocks the action of exonucleases and ligases. The DNA termini become accessible after autophosphorylation of DNA-PKCS, which we demonstrate to require synapsis of DNA ends. Interestingly, the presence of DNA-PK prevents ligation of the two synapsed termini, but allows ligation to another DNA molecule. This alteration of the ligation route is independent of the type of ligase that we used, indicating that the intrinsic architecture of the DNA-PK complex itself is not able to support ligation of the synapsed DNA termini. We present a working model in which DNA-PK creates a stable molecular bridge between two DNA ends that is remodeled after DNA-PK autophosphorylation in such a way that the extreme termini become accessible without disrupting synapsis. We infer that joining of synapsed DNA termini would require an additional protein factor.

Original languageEnglish (US)
Pages (from-to)7238-7246
Number of pages9
JournalNucleic Acids Research
Volume31
Issue number24
DOIs
StatePublished - Dec 15 2003
Externally publishedYes

Fingerprint

DNA-Activated Protein Kinase
Chromosome Pairing
lysyl-arginyl-alanyl-lysyl-alanyl-lysyl-threonyl-threonyl-lysyl-lysyl-arginine
DNA
Ligation
Ligases
DNA End-Joining Repair
Catalytic DNA
Exonucleases
Protein-Serine-Threonine Kinases
Catalytic Domain

ASJC Scopus subject areas

  • Genetics

Cite this

Weterings, E. ., Verkaik, N. S., Brüggenwirth, H. T., Hoeijmakers, J. H. J., & van Gent, D. C. (2003). The role of DNA dependent protein kinase in synapsis of DNA ends. Nucleic Acids Research, 31(24), 7238-7246. https://doi.org/10.1093/nar/gkg889

The role of DNA dependent protein kinase in synapsis of DNA ends. / Weterings, Eric -; Verkaik, Nicole S.; Brüggenwirth, Hennie T.; Hoeijmakers, Jan H J; van Gent, Dik C.

In: Nucleic Acids Research, Vol. 31, No. 24, 15.12.2003, p. 7238-7246.

Research output: Contribution to journalArticle

Weterings, E, Verkaik, NS, Brüggenwirth, HT, Hoeijmakers, JHJ & van Gent, DC 2003, 'The role of DNA dependent protein kinase in synapsis of DNA ends', Nucleic Acids Research, vol. 31, no. 24, pp. 7238-7246. https://doi.org/10.1093/nar/gkg889
Weterings E, Verkaik NS, Brüggenwirth HT, Hoeijmakers JHJ, van Gent DC. The role of DNA dependent protein kinase in synapsis of DNA ends. Nucleic Acids Research. 2003 Dec 15;31(24):7238-7246. https://doi.org/10.1093/nar/gkg889
Weterings, Eric - ; Verkaik, Nicole S. ; Brüggenwirth, Hennie T. ; Hoeijmakers, Jan H J ; van Gent, Dik C. / The role of DNA dependent protein kinase in synapsis of DNA ends. In: Nucleic Acids Research. 2003 ; Vol. 31, No. 24. pp. 7238-7246.
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