(A) Sequence comparison between SLX4TBM and ApolloTBM. Key residues that mediate the interaction with TRF2TRFH are highlighted in red.
(B) ITC measurement of the SLX4TBM-TRF2TRFH interaction. Inset shows the ITC titration.
(C) Overall structure of the SLX4TBM-TRF2TRFH complex. TRF2TRFH and SLX4TBM are colored in green and yellow, respectively, in one monomer, and dark green and orange, respectively, in the other.
(D) Superposition of the TBM peptide-binding sites in the SLX4TBM-TRF2TRFH and ApolloTBM-TRF2TRFH complexes. TRF2TRFH is colored in purple and green in the SLX4TBM-TRF2TRFH and ApolloTBM-TRF2TRFH complexes, respectively.
(E) Equilibrium dissociation constants (Kd) of WT and mutant SLX4TBM-TRF2TRFH interactions measured by ITC. N.D., not detectable.
(F) Co-IP of WT and mutant SLX4 and TRF2 in 293T cells. Lanes marked “IN” contain 5% of the input lysate used for IPs.
(G) Nuclear localization of WT and mutant SLX4 and TRF2. Telomeric DNA was detected by the Cy3-labeled (CCCTAA)3 PNA probe. Bar: 5 mm.
See also Figure S2.