Hanakahi, L. A., Bartlet-Jones, M., Chappell, C., Pappin, D., West, S. C. (September 2000) Binding of inositol phosphate to DNA-PK and stimulation of double-strand break repair. Cell, 102 (6). pp. 721-9. ISSN 0092-8674 (Print)0092-8674 (Linking)
Abstract
In mammalian cells, double-strand breaks in DNA can be repaired by nonhomologous end-joining (NHEJ), a process dependent upon Ku70/80, DNA-PKcs, XRCC4, and DNA ligase IV. Starting with HeLa cell-free extracts, which promote NHEJ in a reaction dependent upon all of these proteins, we have purified a novel factor that stimulates DNA end-joining in vitro. Using a combination of phosphorus NMR, mass spectroscopy, and strong anion exchange chromatography, we identify this factor as inositol hexakisphosphate (IP6). Purified IP6 is bound by DNA-PK and specifically stimulates DNA-PK-dependent end-joining in vitro. The involvement of inositol phosphate in DNA-PK-dependent NHEJ is of particular interest since the catalytic domain of DNA-PKcs is similar to that found in the phosphatidylinositol 3 (PI 3)-kinase family.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | Cell Fractionation Cell-Free System DNA/ metabolism DNA Repair/ physiology DNA-Activated Protein Kinase DNA-Binding Proteins/analysis/isolation & purification/metabolism Genetic Complementation Test HeLa Cells Humans Magnetic Resonance Spectroscopy Mass Spectrometry Nuclear Proteins Phytic Acid/analysis/isolation & purification/ metabolism Protein-Serine-Threonine Kinases/ metabolism Tritium/diagnostic use |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification bioinformatics > genomics and proteomics > genetics & nucleic acid processing bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > DNA binding protein bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > phosphoinositide 3-kinase |
| CSHL Authors: | |
| Communities: | CSHL labs > Pappin lab |
| Depositing User: | Matt Covey |
| Date: | 15 September 2000 |
| Date Deposited: | 31 Jan 2014 16:23 |
| Last Modified: | 31 Jan 2014 16:23 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/29356 |
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