Genomic and Protein Structural Maps of Adaptive Evolution of Human Influenza A Virus to Increased Virulence in the Mouse (original) (raw)
Figure 10
NS1 three dimensional maps of mouse adaptive mutations and effects on CPSF binding.
Mutations are shown as numbered space filling images as described in Fig. 3. (A) in the NS1 monomer; (B) the NS1 dimer effector domain (grey) bound to 2 CPSF F2F3 fragments (dark blue). Amino acid NS1 106 of each monomer is in direct contact in the dimer. CPSF30-F2F3 is in direct contact with NS1 amino acids 103, 106 and 180. (C) Coimmunoprecipitation of HK-wt and V23A, F103L, M106V, M106I, M106I+L98S, and V180A mutant NS1 proteins with FLAG-CPSF30-F2/F3. Recombinant NS1 proteins (2.0 µg) were mixed with equivalent amounts of FLAG-tagged CPSF30-F2/F3 before blotting in parallel using anti-NS1 or anti-FLAG monoclonal antibody respectively to demonstrate the input. Pull down samples were blotted in side-by-side comparisons for immunoglobulin (as a loading standard) and NS1 protein to demonstrate association of NS1 with CPSF30-F2/F3.