View of required ATL2 residues in the prefusion and postfusion conformer. Required residues identified by knockdown replacement (Fig. 1) are shown in cartoon and stick form. The Protein Data Bank coordinates for the prefusion (3QOF) and postfusion (3QNU) ATL1 conformers were downloaded from the RCSB Protein Data Bank database (Bian et al., 2011) and rendered in PyMOL (DeLano Scientific LLC). One subunit is green, and the other is blue. Bound GDP is highlighted in sticks. (A) The location of the three categories of required residues in both pre- and postfusion dimer conformers are boxed and numbered (1–3). (B) A close-up view of the required residues boxed (1–3) in A. Key residues are numbered by their position in the ATL2 sequence and shown in stick form. See the Results under Middle domain mutations fall into three classes for details.