Autophosphorylation of DNA-PKCS regulates its dynamics at DNA double-strand breaks

Naoya Uematsu, Eric Weterings, Ken Ichi Yano, Keiko Morotomi-Yano, Burkhard Jakob, Gisela Taucher-Scholz, Pierre Olivier Mari, Dik C. Van Gent, Benjamin P.C. Chen, David J. Chen

Research output: Contribution to journalArticle

267 Scopus citations

Abstract

The DNA-dependent protein kinase catalytic subunit (DNA-PKCS) plays an important role during the repair of DNA double-strand breaks (DSBs). It is recruited to DNA ends in the early stages of the nonhomologous end-joining (NHEJ) process, which mediates DSB repair. To study DNA-PKCS recruitment in vivo, we used a laser system to introduce DSBs in a specified region of the cell nucleus. We show that DNA-PKCS accumulates at DSB sites in a Ku80-dependent manner, and that neither the kinase activity nor the phosphorylation status of DNA-PKCS influences its initial accumulation. However, impairment of both of these functions results in deficient DSB repair and the maintained presence of DNA-PKCS at unrepaired DSBs. The use of photobleaching techniques allowed us to determine that the kinase activity and phosphorylation status of DNA-PKCS influence the stability of its binding to DNA ends. We suggest a model in which DNA-PKCS phosphorylation/autophosphorylation facilitates NHEJ by destabilizing the interaction of DNA-PKCS with the DNA ends.

Original languageEnglish (US)
Pages (from-to)219-229
Number of pages11
JournalJournal of Cell Biology
Volume177
Issue number2
DOIs
StatePublished - Apr 23 2007

ASJC Scopus subject areas

  • Cell Biology

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    Uematsu, N., Weterings, E., Yano, K. I., Morotomi-Yano, K., Jakob, B., Taucher-Scholz, G., Mari, P. O., Van Gent, D. C., Chen, B. P. C., & Chen, D. J. (2007). Autophosphorylation of DNA-PKCS regulates its dynamics at DNA double-strand breaks. Journal of Cell Biology, 177(2), 219-229. https://doi.org/10.1083/jcb.200608077