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. Author manuscript; available in PMC: 2008 Oct 1.
Published in final edited form as: DNA Repair (Amst). 2007 Jun 12;6(10):1507–1516. doi: 10.1016/j.dnarep.2007.04.014

Figure 7. Direct interaction between XLF/Nej1 and Lig4/Dnl4 in pull-down but not two-hybrid analysis.

Figure 7

(A) Two-hybrid analysis was performed similar to Figure 3 using XLF/Nej1, XRCC4/Lif1 and various fragments of Lig4/Dnl4. Histidine selective indicator medium is shown. In both orientations, a strong XRCC4/Lif1-Lig4/Dnl4 interaction was detected that depended on the C-terminal BRCT domains of Lig4/Dnl4, but no XLF/Nej1-Lig4/Dnl4 interaction was detected. (B) CBP-Nej1, Myc-Lif1 and FLAG-Dnl4 were co-expressed from plasmids as indicated in haploid yeast that carried a chromosomal lif1Δ allele. Pull-down of CBP-Nej1 using calmodulin agarose was followed by immunoblotting for all tags. CBP-Nej1 was able to pull down FLAG-Dnl4 regardless of the presence of Lif1, although with low efficiency. (C) Human proteins were expressed in haploid yeast and pulled down as in (B). Similar to yeast proteins, CBP-XLF was able to pull down FLAG-Lig4 in the absence of XRCC4, although with substantially less efficiency than CBP-XRCC4.