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Mre11 and Ku70 interact in somatic cells, but are differentially expressed in early meiosis

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Abstract

Double-strand DNA breaks (DSBs) pose a major threat to living cells, and several mechanisms for repairing these lesions have evolved. Eukaryotes can process DSBs by homologous recombination (HR) or non-homologous end joining1,2 (NHEJ). NHEJ connects DNA ends irrespective of their sequence, and it predominates in mitotic cells, particularly during G1 ( ref. 3). HR requires interaction of the broken DNA molecule with an intact homologous copy, and allows restoration of the original DNA sequence. HR is active during G2 of the mitotic cycle3 and predominates during meiosis, when the cell creates DSBs (ref. 4), which must be repaired by HR to ensure proper chromosome segregation. How the cell controls the choice between the two repair pathways is not understood. We demonstrate here a physical interaction between mammalian Ku70, which is essential for NHEJ (ref. 5), and Mre11, which functions both in NHEJ and meiotic HR (Refs 2,6). Moreover, we show that irradiated cells deficient for Ku70 are incapable of targeting Mre11 to subnuclear foci that may represent DNA-repair complexes7. Nevertheless, Ku70 and Mre11 were differentially expressed during meiosis. In the mouse testis, Mre11 and Ku70 co-localized in nuclei of somatic cells and in the XY bivalent. In early meiotic prophase, however, when meiotic recombination is most probably initiated8, Mre11 was abundant, whereas Ku70 was not detectable. We propose that Ku70 acts as a switch between the two DSB repair pathways. When present, Ku70 destines DSBs for NHEJ by binding to DNA ends and attracting other factors for NHEJ, including Mre11; when absent, it allows participation of DNA ends and Mre11 in the meiotic HR pathway.

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Figure 1: Fragments of Mre11 and Ku70 proteins used in two-hybrid analyses.
Figure 2: Binding of Ku70 to Mre11 fragments.
Figure 3: Characterization of antibodies and analysis of immune precipitates by western blot.
Figure 4: Immunolocalization of Mre11 and Ku70.
Figure 5: Role of Mre11 and Ku70 in meiotic HR and in NHEJ.

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Acknowledgements

We thank F. Romero and S. Fischer for a cDNA clone encoding the C-terminal part of human Ku70, and P. Pfeiffer for CHO-K1 and xrs-6 cells. This work was supported by the Netherlands Society for Scientific Research NWO (grants no. 900-501-170 and 901-01-097) and the EC (Contracts no. ERBCHRXCT 940511 and BI04 CT97 2028).

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  1. Laboratory of Genetics, Wageningen Agricultural University, , Dreijenlaan 2, Wageningen , 6703 HA, The Netherlands

    Wolfgang Goedecke, Maureen Eijpe, Hildo H. Offenberg, Mirjam van Aalderen & Christa Heyting

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  1. Wolfgang Goedecke
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  2. Maureen Eijpe
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  3. Hildo H. Offenberg
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  5. Christa Heyting
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Correspondence to Christa Heyting.

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Goedecke, W., Eijpe, M., Offenberg, H. et al. Mre11 and Ku70 interact in somatic cells, but are differentially expressed in early meiosis. Nat Genet 23, 194–198 (1999). https://doi.org/10.1038/13821

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