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  • Original Paper
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Inhibition of Ku heterodimer DNA end binding activity during granulocytic differentiation of human promyelocytic cell lines

Oncogene volume 20, pages 4373–4382 (2001)Cite this article

Abstract

The heterodimeric Ku protein (composed of the Ku 86 and Ku 70 sub-units) is a nuclear protein which binds to DNA termini without sequence specificity. Ku is the DNA-targeting component of the large catalytic sub-unit of the DNA-dependent protein kinase complex that is required for the repair of DNA double-strand breaks in mammalian cells. We studied the expression and function of Ku/DNA-PK during granulocytic differentiation of two human promyelocytic cell lines, HL60 and NB4, a process associated to decreased radiation resistance. After 3 days exposure to differentiating agents (either all-trans-retinoic acid or DMSO), Ku binding to double stranded (ds)-DNA ends declined dramatically whereas Ku protein levels remain unchanged. The nuclear, but not cytoplasmic, fraction of differentiated HL60 cells extracts exhibited a heat-sensitive inhibitory activity towards DNA binding of recombinant Ku heterodimer. We further demonstrate that immunoprecipitation of Ku is impaired in extracts from differentiated cells by using two antibodies that recognize epitopes within the C-terminus DNA binding domains of Ku 70 and Ku 86 proteins. These results favor the hypothesis of a protein interacting with Ku that would prevent DNA binding of heterodimerized Ku protein by steric hindrance.

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Acknowledgements

This study was supported by grants from the ‘Ligue Regionale Contre le Cancer coruté Midi-Pyrenees’, the ‘Association de Recherche Contre le Cancer’ and the ‘Comité de Radioprotection d'Electricité Gaz de France’.

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  1. Institut de Pharmacologie et de Biologie Structurale (CNRS, UPR 9062) 205 route de Narbonne, Toulouse cedex, 31077, France

    Catherine Muller, Sylvie Monferran, Alexander-Christopher Gamp, Patrick Calsou & Bernard Salles

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  1. Catherine Muller
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  2. Sylvie Monferran
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Correspondence to Bernard Salles.

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Muller, C., Monferran, S., Gamp, AC. et al. Inhibition of Ku heterodimer DNA end binding activity during granulocytic differentiation of human promyelocytic cell lines. Oncogene 20, 4373–4382 (2001). https://doi.org/10.1038/sj.onc.1204571

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