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

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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

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DNA-Activated Protein Kinase
Double-Stranded DNA Breaks
Catalytic Domain
Phosphorylation
DNA
Phosphotransferases
Photobleaching
Cell Nucleus
Lasers

ASJC Scopus subject areas

  • Cell Biology

Cite this

Uematsu, N., Weterings, E., Yano, K. I., Morotomi-Yano, K., Jakob, B., Taucher-Scholz, G., ... 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

Autophosphorylation of DNA-PKCS regulates its dynamics at DNA double-strand breaks. / Uematsu, Naoya; Weterings, Eric; Yano, Ken Ichi; Morotomi-Yano, Keiko; Jakob, Burkhard; Taucher-Scholz, Gisela; Mari, Pierre Olivier; Van Gent, Dik C.; Chen, Benjamin P C; Chen, David J.

In: Journal of Cell Biology, Vol. 177, No. 2, 23.04.2007, p. 219-229.

Research output: Contribution to journalArticle

Uematsu, N, Weterings, E, Yano, KI, Morotomi-Yano, K, Jakob, B, Taucher-Scholz, G, Mari, PO, Van Gent, DC, Chen, BPC & Chen, DJ 2007, 'Autophosphorylation of DNA-PKCS regulates its dynamics at DNA double-strand breaks', Journal of Cell Biology, vol. 177, no. 2, pp. 219-229. https://doi.org/10.1083/jcb.200608077
Uematsu N, Weterings E, Yano KI, Morotomi-Yano K, Jakob B, Taucher-Scholz G et al. Autophosphorylation of DNA-PKCS regulates its dynamics at DNA double-strand breaks. Journal of Cell Biology. 2007 Apr 23;177(2):219-229. https://doi.org/10.1083/jcb.200608077
Uematsu, Naoya ; Weterings, Eric ; Yano, Ken Ichi ; Morotomi-Yano, Keiko ; Jakob, Burkhard ; Taucher-Scholz, Gisela ; Mari, Pierre Olivier ; Van Gent, Dik C. ; Chen, Benjamin P C ; Chen, David J. / Autophosphorylation of DNA-PKCS regulates its dynamics at DNA double-strand breaks. In: Journal of Cell Biology. 2007 ; Vol. 177, No. 2. pp. 219-229.
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