Atg16L1-FIP200 interacting domains. (A) Mapping of the Atg16L1-binding site on FIP200. Yeast two-hybrid mapping experiments showing that Atg16L1 binds to both the N-terminal and C-terminal coiled-coil regions of FIP200. (B) Mapping of the FIP200-binding site within Atg16L1. Yeast two-hybrid mapping experiments showing that the 1–840 fragment of FIP200 binds to the C-terminal 247–607 region, while the 1276–1591 fragment of FIP200 binds to the N-terminal 1–264 region of Atg16L1. (C and D) Additional yeast two-hybrid mapping experiments showing that the binding site of the FIP200 1276–1591 fragment maps to residues 239–246 of Atg16L1. White lines divide images derived from the same plate, and red lines divide images from different plates. (E) The N-terminal 1–249 fragment of Atg16L1 is sufficient to interact with FIP200. Full-length (lanes 1 and 2) or the N-terminal 1–249 fragments (lanes 3 and 4) of Myc-Atg16L1 coprecipitations with empty vector controls (lanes 1 and 3) or OSF-FIP200 1276–1591 (lanes 2 and 4). (F) Co-precipitation experiments confirming that OSF-FIP200 1276–1591 coprecipitates with wild-type (WT) Myc-Atg16L1 (1–249) fragments, but not with analogous fragments containing a mutant FIP200-binding site (Myc-Atg16L1 ΔWD 239–242A).