Specific interaction of IP6 with human Ku70/80, the DNA-binding subunit of DNA-PK

被引:87
作者
Hanakahi, LA [1 ]
West, SC [1 ]
机构
[1] Canc Res UK, London Res Inst, Clare Hall Labs, S Mimms EN6 3LD, Herts, England
关键词
DNA repair; double-strand breaks; non-homologous end-joining; protein kinase;
D O I
10.1093/emboj/21.8.2038
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In eukaryotic cells, DNA double-strand breaks can be repaired by non-homologous end-joining, a process dependent upon Ku70/80, XRCC4 and DNA ligase IV. In mammals, this process also requires DNA-PKcs, the catalytic subunit of the DNA-dependent protein kinase DNA-PK. Previously, inositol hexakisphosphate (IP6) was shown to be bound by DNA-PK and to stimulate DNA-PK-dependent end-joining in vitro. Here, we localize IP6 binding to the Ku70/80 subunits of DNA-PK, and show that DNA-PK, alone exhibits no detectable affinity for IP6. Moreover, proteolysis mapping of Ku70/80 in the presence and absence of IP6 indicates that binding alters the conformation of the Ku70/80 heterodimer. The yeast homologue of Ku70/80, yKu70/80, fails to bind IP6, indicating that the function of IP6 in non-homologous end-joining, like that of DNA-PKcs, is unique to the mammalian end-joining process.
引用
收藏
页码:2038 / 2044
页数:7
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