Figure 3.

Mutational limits of the Rev ARM and NES. (A) CDMS fitness values for residues in the Rev ARM¹⁰ (B) Structure of Rev NES (yellow) bound to Crm1 (grey) (PDBID: 3NBZ) with the Rev hydrophobic residues binding to the Crm1 NES binding groove shown as spheres (C) CDMS fitness values for the NES¹⁰ (D) Preferences for Rev NES contact residues using CDMS fitness values¹⁰. (E) Reporter assay probing export activity of Rev NES mutations. Data are mean ± standard deviation (s.d) of biological replicates. Western blots below show protein expression for Strep-tagged Rev and GAPDH as loading control, and numbers above blots represent quantified band intensities for Rev mutants relative to WT-Rev. The original blot for the cropped bands is shown in Fig. S5. (F) Viral replication profiles of viruses containing Rev with NES mutations. Data are mean ± s.d of biological triplicates.