Figure 6

Mutating positive residues in the MTase αE helix prevents WNV MTase’s interaction with PKG. In the WNV MTase, Lys189, Lys193, Arg199, and Arg200 (Table 1) were mutated to Ala to create WNV MTase (mut). HEK293T cells were transfected with plasmids encoding WNV MTase (WT) or WNV MTase (mut) and PKGIβ-Flag or were mock transfected. PKGIβ was immunoprecipitated using α-Flag, and the presence of WNV MTase in co-immunoprecipitation and lysate samples was detected by Western blot with α-WNV MTase. PKG and actin were detected by Western blotting with α-Flag and α-actin, respectively.