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. 2018 Feb 12;92(5):e02097-17. doi: 10.1128/JVI.02097-17

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Copyright © 2018 Turkington et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

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FIG 4 — Engineering the bat FLUAV (Guat/09) NS1 protein to bind p85β. (A) Crystal structure of the PR8/NS1 ED in complex with part of the p85β iSH2 domain. The NS1 ED is shown in gray, with p85β contact residues highlighted in yellow. The p85β iSH2 domain is colored blue. The six NS1 amino acid residues chosen for mutagenesis studies (see panel B, Guat/09 numbering) are highlighted in red. The figure was generated using PyMOL with PDB 3L4Q. (B) Comparison of key NS1 ED residues between human (PR8, Cal09), swine (Sw/Tx/98), avian (Nig/07, Sh/13), and bat (Guat/09, Peru/10) NS1 proteins. #, Guat/09 numbering. (C and D) 293T cells were cotransfected for 48 h with plasmids expressing the indicated V5-tagged NS1 protein (or GST) and FLAG-tagged human p85β. After cell lysis, clarification, and anti-V5 immunoprecipitation, soluble (input) and pulldown (IP:α-V5) fractions were analyzed by SDS-PAGE and Western blotting. Data are representative of at least two independent experiments.