Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
DNA-dependent protein kinase activity is absent in xrs-6 cells: implications for site-specific recombination and DNA double-strand break repair.
Finnie NJ
,
Gottlieb TM
,
Blunt T
,
Jeggo PA
,
Jackson SP
.
???displayArticle.abstract???
DNA-dependent protein kinase (DNA-PK) is a nuclear serine/threonine protein kinase composed of a catalytic subunit called p350 and a DNA binding component termed Ku. Ku consists of two tightly associated polypeptides of approximately 70 kDa and 80 kDa (Ku80). An intriguing feature of DNA-PK is that it binds to DNA ends and other discontinuities in DNA and requires these structures for its activation. This suggests that DNA-PK may function in DNA repair and/or recombination. Consistent with this, Ku DNA binding activity was shown recently to be absent in extracts of hamster xrs-6 cells, which are defective in DNA double-strand (ds) break repair and V(D)J recombination. Furthermore, xrs-6 cells are complemented by expression of the Ku80 cDNA. To date, DNA-PK activity has been demonstrated unequivocally only in extracts of primate cells. Here, we describe an assay that can detect DNA-PK activity in extracts of mouse, hamster, Xenopus, and Drosophila cells. Using this assay, we find that xrs-6 cells completely lack DNA-PK activity. By contrast, xrs-6 derivatives complemented by human chromosome fragments bearing the Ku80 gene have restored both the DNA end binding and kinase activities associated with DNA-PK. Finally, we show that xrs-6 extracts are complemented biochemically by purified Ku. Our findings indicate that the xrs-6 defects are direct consequences of the mutation in Ku80 and implicate DNA-PK in recombination and DNA repair processes.
Anderson,
DNA damage and the DNA-activated protein kinase.
1993, Pubmed
Anderson,
DNA damage and the DNA-activated protein kinase.
1993,
Pubmed
Anderson,
The nuclear serine/threonine protein kinase DNA-PK.
1992,
Pubmed
Bannister,
c-Jun is phosphorylated by the DNA-dependent protein kinase in vitro; definition of the minimal kinase recognition motif.
1993,
Pubmed
Cai,
Chromosomal location and expression of the genes coding for Ku p70 and p80 in human cell lines and normal tissues.
1994,
Pubmed
Carter,
A DNA-activated protein kinase from HeLa cell nuclei.
1990,
Pubmed
DePamphili,
How transcription factors regulate origins of DNA replication in eukaryotic cells.
1993,
Pubmed
Drapkin,
Where transcription meets repair.
1994,
Pubmed
Dröge,
Transcription-driven site-specific DNA recombination in vitro.
1993,
Pubmed
Dvir,
Purification and characterization of a template-associated protein kinase that phosphorylates RNA polymerase II.
1993,
Pubmed
Falzon,
The nucleotide sequence of a mouse cDNA encoding the 80 kDa subunit of the Ku (p70/p80) autoantigen.
1992,
Pubmed
Feldmann,
A putative homologue of the human autoantigen Ku from Saccharomyces cerevisiae.
1993,
Pubmed
Finnie,
Transcription factor phosphorylation by the DNA-dependent protein kinase.
1993,
Pubmed
Getts,
Absence of a Ku-like DNA end binding activity in the xrs double-strand DNA repair-deficient mutant.
1994,
Pubmed
Gottlieb,
The DNA-dependent protein kinase: requirement for DNA ends and association with Ku antigen.
1993,
Pubmed
Gottlieb,
Protein kinases and DNA damage.
1994,
Pubmed
Hafezparast,
Subchromosomal localization of a gene (XRCC5) involved in double strand break repair to the region 2q34-36.
1993,
Pubmed
Jackson,
Regulating transcription factor activity by phosphorylation.
1992,
Pubmed
Jackson,
GC box binding induces phosphorylation of Sp1 by a DNA-dependent protein kinase.
1990,
Pubmed
Jacoby,
Yolk protein factor 1 is a Drosophila homolog of Ku, the DNA-binding subunit of a DNA-dependent protein kinase from humans.
1994,
Pubmed
Jeggo,
X-ray-sensitive mutants of Chinese hamster ovary cell line. Isolation and cross-sensitivity to other DNA-damaging agents.
1983,
Pubmed
Jeggo,
Localization of a DNA repair gene (XRCC5) involved in double-strand-break rejoining to human chromosome 2.
1992,
Pubmed
Jeggo,
Genetic analysis of ionising radiation sensitive mutants of cultured mammalian cell lines.
1991,
Pubmed
Kemp,
X-ray sensitive mutants of Chinese hamster ovary cells defective in double-strand break rejoining.
1984,
Pubmed
Lees-Miller,
Human cells contain a DNA-activated protein kinase that phosphorylates simian virus 40 T antigen, mouse p53, and the human Ku autoantigen.
1990,
Pubmed
Lees-Miller,
Human DNA-activated protein kinase phosphorylates serines 15 and 37 in the amino-terminal transactivation domain of human p53.
1992,
Pubmed
Leung,
Transcriptional regulatory elements stimulate recombination in extrachromosomal substrates carrying immunoglobulin switch-region sequences.
1992,
Pubmed
Miller,
Comparison of in vivo and in vitro ribosomal RNA synthesis in nucleolar mutants of Xenopus laevis.
1977,
Pubmed
,
Xenbase
Morozov,
DNA-dependent protein kinase is activated by nicks and larger single-stranded gaps.
1994,
Pubmed
Porges,
Antigenic determinants of the Ku (p70/p80) autoantigen are poorly conserved between species.
1990,
Pubmed
Rathmell,
Involvement of the Ku autoantigen in the cellular response to DNA double-strand breaks.
1994,
Pubmed
Schöler,
A family of octamer-specific proteins present during mouse embryogenesis: evidence for germline-specific expression of an Oct factor.
1989,
Pubmed
Taccioli,
Impairment of V(D)J recombination in double-strand break repair mutants.
1993,
Pubmed
Taccioli,
A DNA repair defect in Chinese hamster ovary cells affects V(D)J recombination similarly to the murine scid mutation.
1994,
Pubmed
Taccioli,
Ku80: product of the XRCC5 gene and its role in DNA repair and V(D)J recombination.
1994,
Pubmed
Thacker,
The genetic basis of resistance to ionising radiation damage in cultured mammalian cells.
1991,
Pubmed
Wang,
Phosphorylation sites in the amino-terminal region of mouse p53.
1992,
Pubmed
