Wac directly interacts with Dgt2 via its coiled-coil region. (A) A series of Wac truncations were translated in vitro in reticulocyte lysate in the presence of [³⁵S]-methionine (input), and interaction with bacterially produced MBP-Dgt2 (Dgt2) or MBP were tested by pull-down. (B) Consecutive residues within the coiled-coil region of Wac were systematically replaced with alanines, and the interaction with Dgt2 was tested. (C) A sequence comparison of Wac homologues among insects. Red letters indicate residues identical to those in D. melanogaster, and the asterisks indicate residues essential for the interaction. Dmel, D. melanogaster; Dana, Drosophila ananassae; Dpse, Drosophila pseudoobscura; Dvir, Drosophila virilis; Agam, Anopheles gambiae; Aaeg, Aedes aegypti.