DNA-dependent protein kinase is activated by nicks and larger single-stranded gaps

Viktor E. Morozov, Miriam Falzon, Carl W. Anderson, Edward L. Kuff

Research output: Contribution to journalArticle

126 Citations (Scopus)

Abstract

DNA-PK is a DNA-activated serine/threonine protein kinase capable of phosphorylating a number of nuclear DNA-binding proteins. Purified human DNA-PK has two subunits, a 350-kDa polypeptide, Prkdc, which binds ATP and is presumed to contain the catalytic site, and the Ku autoantigen which mediates DNA binding and activation. Previous studies have shown that DNA-PK is activated in vitro by linear double-stranded DNA fragments; however, the Ku subunit binds a broader range of DNA structures. Here we show that EBP-80, a protein originally isolated as a transcription factor for a retroviral long terminal repeat element and subsequently found to be similar to if not identical with Ku, also mediates kinase activation. The EBP-80-Prkdc complex is activated by nanomolar concentrations of DNA constructs containing single-to-double strand transitions, including a closed stem-loop structure and single strand gaps of 0 (nick), 6, and 30 nucleotides. Kinase activation parallels the ability of EBP-80 to bind these and other constructs. Our results extend the range of DNA configurations known to activate DNA-PK and are consistent with the participation of this enzyme complex in several nuclear functions.

Original languageEnglish (US)
Pages (from-to)16684-16688
Number of pages5
JournalJournal of Biological Chemistry
Volume269
Issue number24
StatePublished - Jun 17 1994

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DNA-Activated Protein Kinase
DNA
Chemical activation
Phosphotransferases
Terminal Repeat Sequences
Protein-Serine-Threonine Kinases
DNA-Binding Proteins
Nuclear Proteins
Autoantigens
Catalytic Domain
Transcription Factors
Nucleotides
Adenosine Triphosphate

ASJC Scopus subject areas

  • Biochemistry

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DNA-dependent protein kinase is activated by nicks and larger single-stranded gaps. / Morozov, Viktor E.; Falzon, Miriam; Anderson, Carl W.; Kuff, Edward L.

In: Journal of Biological Chemistry, Vol. 269, No. 24, 17.06.1994, p. 16684-16688.

Research output: Contribution to journalArticle

Morozov, VE, Falzon, M, Anderson, CW & Kuff, EL 1994, 'DNA-dependent protein kinase is activated by nicks and larger single-stranded gaps', Journal of Biological Chemistry, vol. 269, no. 24, pp. 16684-16688.
Morozov, Viktor E. ; Falzon, Miriam ; Anderson, Carl W. ; Kuff, Edward L. / DNA-dependent protein kinase is activated by nicks and larger single-stranded gaps. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 24. pp. 16684-16688.
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