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Spagnolo L, Rivera-Calzada A, Pearl LH, Llorca O (2006) Three-dimensional structure of the human dna-pkcs/ku70/ku80 complex assembled on DNA and its implications for DNA dsb repair. Mol Cell 22(4):511–519 78. Suchanek M, Radzikowska A, Thiele C (2005) Photo-leucine and photo-methionine allow identification of protein-protein interactions in living cells. Nat Methods 2(4):261–267. 1038/nmeth752 79. Svergun DI, Koch MHJ (2003) Small-angle scattering studies of biological macromolecules in solution.

GraFix, ultracentrifugation in solutions with glycerol and fixation gradients is a recently developed method to provide homogeneous samples, and can improve the quality of EM images [40]. Model building into EM maps can be achieved with and without crystallographic structures [46, 68]. As a guide of the modeling, hexahistidine tags and DNA labeled with gold clusters (Nanoprobes) allow the location of the tags and DNA in complexes. In reverse, EM maps have been successfully used for phasing X-ray diffraction data by molecular replacement [51, 88].

The region of the XRCC4 C-terminal domain after residue 213 is not included in current solved XRCC4 crystal structures due to its predicted flexible structure. XRCC4 can exist as a salt-dependent equilibrium of dimers and tetramers in solution [49]. The dynamic of XRCC4 oligomers formation can be also shifted to dimmers through strong binding of LigIV to XRCC4 C-terminal helices [49]. The binding region between XRCC4 and LigIV overlaps with the XRCC4 tetramerisation region, which may explain why LigIV functions as a strong competitor to shift the equilibrium towards the XRCC4 dimer in solution [49] (Fig.

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