Binding of inositol phosphate to DNA-PK and stimulation of double-strand break repair

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)

URL: http://www.ncbi.nlm.nih.gov/pubmed/11030616
DOI: 10.1016/S0092-8674(00)00061-1

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: http://repository.cshl.edu/id/eprint/29356

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