Priestley, A (1998) Molecular and biochemical characterisation of DNA-dependent protein kinase-defective rodent mutant irs-20. Nucleic Acids Research, 26 (8). pp. 1965-1973. DOI https://doi.org/10.1093/nar/26.8.1965
Priestley, A (1998) Molecular and biochemical characterisation of DNA-dependent protein kinase-defective rodent mutant irs-20. Nucleic Acids Research, 26 (8). pp. 1965-1973. DOI https://doi.org/10.1093/nar/26.8.1965
Priestley, A (1998) Molecular and biochemical characterisation of DNA-dependent protein kinase-defective rodent mutant irs-20. Nucleic Acids Research, 26 (8). pp. 1965-1973. DOI https://doi.org/10.1093/nar/26.8.1965
Abstract
The catalytic subunit of the DNA-dependent protein kinase (DNA-PKcs) is a member of a sub-family of phosphatidylinositol (PI) 3-kinases termed PIK-related kinases. A distinguishing feature of this sub-family is the presence of a conserved C-terminal region downstream of a PI 3-kinase domain. Mutants defective in DNA-PKcs are sensitive to ionising radiation and are unable to carry out V(D)J recombination. Irs-20 is a DNA-PKcs-defective cell line with milder γ-ray sensitivity than two previously characterised mutants, V-3 and mouse scid cells. Here we show that the DNA-PKcs protein from irs-20 cells can bind to DNA but is unable to function as a protein kinase. To verify the defect in irs-20 cells and provide insight into the function and expression of DNA-PKcs in double-strand break repair and V(D)J recombination we introduced YACs encoding human and mouse DNA-PKcs into defective mutants and achieved complementation of the defective phenotypes. Furthermore, in irs-20 we identified a mutation in DNA-PKcs that causes substitution of a lysine for a glutamic acid in the fourth residue from the C-terminus. This represents a strong candidate for the inactivating mutation and provides supportive evidence that the extreme C-terminal motif is important for protein kinase activity.
Item Type: | Article |
---|---|
Uncontrolled Keywords: | Cell Line; CHO Cells; Chromosomes, Artificial, Yeast; Animals; Horses; Humans; Mice; Mice, SCID; DNA Damage; VDJ Recombinases; DNA Nucleotidyltransferases; DNA-Binding Proteins; Nuclear Proteins; DNA; Transfection; Polymerase Chain Reaction; Dose-Response Relationship, Radiation; Cell Survival; DNA Repair; Gene Library; Gamma Rays; Cricetinae; DNA-Activated Protein Kinase; Phosphatidylinositol 3-Kinases; Protein Serine-Threonine Kinases |
Subjects: | Q Science > QH Natural history > QH426 Genetics |
Divisions: | Faculty of Science and Health Faculty of Science and Health > Life Sciences, School of |
SWORD Depositor: | Unnamed user with email elements@essex.ac.uk |
Depositing User: | Unnamed user with email elements@essex.ac.uk |
Date Deposited: | 23 Oct 2014 12:14 |
Last Modified: | 07 Aug 2024 20:26 |
URI: | http://repository.essex.ac.uk/id/eprint/11096 |