Fig. 1. Domain Analysis of RTEL1.

a Schematic representation of RTEL1 proteins and constructs used in this study. The location of the RTEL1^R1264H mutation is indicated with a green star and G4 binding is illustrated. b RTEL1 interacts with itself in cells and the RTEL1^R1264H mutation disrupts interactions within the RING domain. Left, Myc immunoprecipitations from HEK293T cells co-expressing FLAG and myc-tagged RTEL1 (FL) and RTEL1^R1264H (FL ^RH) were carried out. Immunoblotting with FLAG indicates co-IP of both RTEL1 and RTEL1^R1264H. Center, immunoblotting with myc indicates co-IP of RTEL1^∆1097 (∆1097), but not RTEL1^{∆1097 R1264H} (∆1097 ^RH) with GFP-tagged RTEL1 and RTEL1^R1264H. Right, immunoblotting with FLAG shows co-IP of RTEL1^∆1097, but not RTEL1^{∆1097R1264H}. Molecular weight markers, KDa are shown. All immunoblotting experiments were repeated at least three times with similar results. c Oligomeric states of RTEL1 proteins were determined by SEC-MALS analysis of RTEL1^∆762 (blue), RTEL1^∆762R1264H (red), and RTEL1^∆762-1097 (green) at 150 and 300 mM NaCl. Normalized A280 are shown and the calculated molecular mass is shown as a line in the corresponding color across each peak with the secondary scale on the right. The expected and calculated molecular weights are shown (see also Supplementary Fig. 1).