DNA-PKcs promotes chromatin decondensation to facilitate initiation of the DNA damage response

Huiming Lu, Janapriya Saha, Pauline J. Beckmann, Eric A. Hendrickson, Anthony J. Davis

Research output: Contribution to journalArticle

Abstract

The DNA damage response (DDR) encompasses the cellular response to DNA double-stranded breaks (DSBs), and includes recognition of the DSB, recruitment of numerous factors to the DNA damage site, initiation of signaling cascades, chromatin remodeling, cell-cycle checkpoint activation, and repair of the DSB. Key drivers of the DDR are multiple members of the phosphatidylinositol 3-kinase-related kinase family, including ataxia telangiectasia mutated (ATM), ataxia telangiectasia and Rad3-related (ATR), and the DNA-dependent protein kinase catalytic subunit (DNA-PKcs). ATM and ATR modulate multiple portions of the DDR, but DNA-PKcs is believed to primarily function in the DSB repair pathway, non-homologous end joining. Utilizing a human cell line in which the kinase domain of DNA-PKcs is inactivated, we show here that DNA-PKcs kinase activity is required for the cellular response to DSBs immediately after their induction. Specifically, DNA-PKcs kinase activity initiates phosphorylation of the chromatin factors H2AX and KAP1 following ionizing radiation exposure and drives local chromatin decondensation near the DSB site. Furthermore, loss of DNA-PKcs kinase activity results in a marked decrease in the recruitment of numerous members of the DDR machinery to DSBs. Collectively, these results provide clear evidence that DNA-PKcs activity is pivotal for the initiation of the DDR.

Original languageEnglish (US)
Pages (from-to)9467-9479
Number of pages13
JournalNucleic acids research
Volume47
Issue number18
DOIs
StatePublished - Oct 10 2019

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DNA-Activated Protein Kinase
DNA Damage
Chromatin
Catalytic Domain
Ataxia Telangiectasia
Phosphotransferases
Phosphatidylinositol 3-Kinase
Chromatin Assembly and Disassembly
Double-Stranded DNA Breaks
Cell Cycle Checkpoints
Ionizing Radiation
Phosphorylation
Cell Line

PubMed: MeSH publication types

  • Journal Article
  • Research Support, N.I.H., Extramural

Cite this

DNA-PKcs promotes chromatin decondensation to facilitate initiation of the DNA damage response. / Lu, Huiming; Saha, Janapriya; Beckmann, Pauline J.; Hendrickson, Eric A.; Davis, Anthony J.

In: Nucleic acids research, Vol. 47, No. 18, 10.10.2019, p. 9467-9479.

Research output: Contribution to journalArticle

Lu, Huiming ; Saha, Janapriya ; Beckmann, Pauline J. ; Hendrickson, Eric A. ; Davis, Anthony J. / DNA-PKcs promotes chromatin decondensation to facilitate initiation of the DNA damage response. In: Nucleic acids research. 2019 ; Vol. 47, No. 18. pp. 9467-9479.
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