Residues in Loop L2 and the interdomain linker contribute to dmDcr1 binding. (A) Schematic representation of GST-tagged proteins used for the pulldown assays. Positions of mutated amino acids are indicated with red lines. (B) Pulldown assay of His-dmDcr1 from SF21 cell lysate with GST-Loqs-dsRBD3 and point mutants as indicated. GST alone served as control. Bound dmDcr1 is detected by immunoblot using a specific antibody and bait proteins were stained with Coomassie blue. (C) Pulldown assay as in B using Loqs constructs (Loqs0-4) containing sequential truncations of the interdomain linker. (D) GST-pulldown with various mutations within the minimal linker Loqs-3 construct and quantification of the dmDcr1 signal normalized to the wt construct.
