Human ku autoantigen binds cisplatin damaged dna but fails to stimulate dna-activated protein kinase

John Turchi, Karen Henkels

Research output: Contribution to journalArticle

Abstract

We have identified a series of proteins based on an affinity for cisplatin damaged DNA. One protein termed DRP-1 has been purified to homogeneity and was isolated as two distinct complexes. The first complex is a heterodimer of 83 and 68 kDa subunits, while the second complex is a heterotrimer of 350, 83 and 68 kDa subunits in a 1:1:1 ratio. The 83 and 68 kDa subunits in each complex are identical. The 83 kDa subunit of DRP-1 was identified as the p80 subunit of Ku autoantigen by N-terminal protein sequence and western blot analysis. The 68 kDa subunit of DRP-1 cross reacted with monoclonal antisera raised against the Ku autoantigen p70 subunit. The 350 kDa subunit was identified as Prkdc, the catalytic subunit of the human DNA-activated protein kinase, DNA-PK. DRP-1 DNA binding was assessed in mobility shift assays and results indicate that DNA binding was essentially unaffected by cisplatin-DNA adducts in the presence or absence of Prkdc. DNA-PK activity was only stimulated with undamaged DNA, despite the ability of Ku to bind to cisplatin damaged DNA. The lack of DNA-PK stimulation by cisplatin damaged DNA correlated with the extent of cisplatin-DNA adduct formation. These results are discussed with respect to the repair of cisplatin-DNA adducts and the role of DNA-PK in coordinating DNA repair processes.

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

Fingerprint

autoantigens
cisplatin
Autoantigens
protein kinases
Protein Kinases
Cisplatin
DNA
DNA adducts
Repair
Ku Autoantigen
DNA-Activated Protein Kinase
Proteins
Electrophoretic Mobility Shift Assay
protein subunits
DNA Repair
DNA repair
Immune Sera
Catalytic Domain
antiserum
Assays

ASJC Scopus subject areas

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

Cite this

Human ku autoantigen binds cisplatin damaged dna but fails to stimulate dna-activated protein kinase. / Turchi, John; Henkels, Karen.

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

Research output: Contribution to journalArticle

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