The dna double-strand break repair mutant cell line irs-20 expresses an inactive dna-dependent protein kinase catalytic subunit

Scott R. Peterson, Murray Stackhouse, Fanqing Chen, E. Morton Bradbury, David J. Chen

Research output: Contribution to journalArticle

Abstract

The DNA-dependent protein kinase (DNA-PK) is a trimeric enzyme consisting of a 460kD catalytic subunit (DNA-PKcs) and a heterodimeric DNA-binding complex comprised of the 70 and 86 kD Ku autoantigen proteins. Mutations that affect the expression of the Ku 80 or catalytic subunit of DNA-PK (DNA-PKcs) disrupt V(D)J recombination and DNA double-strand break repair pathways. Here, we show the radiosensitive rodent cell line IRS-20 contains a defect that disrupts the expression and kinase activity of DNA-PK. Expression of the DNA-PKcs is reduced four-fold in the IRS-20 cells relative to the parental cell 10B2 and the IRS-20 enzyme is inactive using recombinant RPA as an in vitro substrate for kinase activity. Human chromosome 8, which contains the gene encoding the human DNA-PKcs, complements the radiosensitivity defect of the IRS-20 cells and restores DNA-PK kinase activity. These results suggest the kinase activity of DNA-PK is neccessary for DNA double-stranded break repair and support a model whereby DNA-PK regulates double-stranded DNA break repair via the phosphorylation of protein substrates, some of which may directly participate in this DNA-repair pathway.

Original languageEnglish (US)
JournalFASEB Journal
Volume10
Issue number6
StatePublished - 1996

Fingerprint

DNA-Activated Protein Kinase
protein subunits
protein kinases
Protein Kinases
Catalytic Domain
Repair
Cells
cell lines
Cell Line
mutants
Double-Stranded DNA Breaks
DNA
Phosphotransferases
Catalytic DNA
DNA Repair
V(D)J Recombination
phosphotransferases (kinases)
Chromosomes, Human, Pair 8
Radiation Tolerance
Human Chromosomes

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Peterson, S. R., Stackhouse, M., Chen, F., Morton Bradbury, E., & Chen, D. J. (1996). The dna double-strand break repair mutant cell line irs-20 expresses an inactive dna-dependent protein kinase catalytic subunit. FASEB Journal, 10(6).

The dna double-strand break repair mutant cell line irs-20 expresses an inactive dna-dependent protein kinase catalytic subunit. / Peterson, Scott R.; Stackhouse, Murray; Chen, Fanqing; Morton Bradbury, E.; Chen, David J.

In: FASEB Journal, Vol. 10, No. 6, 1996.

Research output: Contribution to journalArticle

Peterson, SR, Stackhouse, M, Chen, F, Morton Bradbury, E & Chen, DJ 1996, 'The dna double-strand break repair mutant cell line irs-20 expresses an inactive dna-dependent protein kinase catalytic subunit', FASEB Journal, vol. 10, no. 6.
Peterson, Scott R. ; Stackhouse, Murray ; Chen, Fanqing ; Morton Bradbury, E. ; Chen, David J. / The dna double-strand break repair mutant cell line irs-20 expresses an inactive dna-dependent protein kinase catalytic subunit. In: FASEB Journal. 1996 ; Vol. 10, No. 6.
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