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Papers In Press, published online ahead of print February 20, 2001
J. Biol. Chem, 10.1074/jbc.M100768200
Submitted on January 26, 2001
Revised on February 12, 2001
Accepted on February 20, 2001
Hormones, Growth and Development, The Loeb Health Research Institute, Ottawa, Ontario K1Y 4K9
Corresponding Author: rhache{at}lri.ca
The Ku antigen (Ku70/Ku80) is a regulatory subunit of DNA-dependent protein kinase (DNA-PK) that promotes the recruitment of the catalytic subunit of DNA-PK (DNA-PKcs) to DNA-ends and to specific DNA sequences from which the kinase is activated. Ku/DNA-PKcs play essential roles in double-stranded DNA-break repair, V(D)J recombination and have been implicated in the regulation of specific gene transcription. In a yeast two-hybrid screen of a Jurkat T cell cDNA library we have identified a specific interaction between the 70 kDa subunit of Ku heterodimer and the homeodomain of HOXC4, a homeodomain protein expressed in the hematopoetic system. Unexpectedly, a similar interaction with Ku was observed for several additional homeodomain proteins including octamer transcription factors 1 and 2, and Dlx2, suggesting that specific binding to Ku may be a property shared by many homeodomain proteins. Ku-homeodomain binding was mediated through the extreme C-terminus of Ku70 and was abrogated by amino acid substitutions at K595/K596. Ku binding allowed the recruitment of the homeodomain to DNA-ends and dramatically enhanced the phosphorylation of homeodomain-containing proteins by DNA-PK. These results suggest that Ku functions as a substrate docking protein for signaling by DNA-PK to homeodomain proteins from DNA ends.
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