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. 2016 Mar;22(3):383–396. doi: 10.1261/rna.055426.115

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© 2016 Jakob et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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FIGURE 2. — 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.