Figure 5: Residues in the S subunit that are important for particle assembly and genome encapsidation.
From: Mechanisms of assembly and genome packaging in an RNA virus revealed by high-resolution cryo-EM
(a) Agarose gels stained with either Coomassie blue or ethidium bromide (EtBr) show that the F192W mutant packages no RNA. Negative stain EM illustrates ‘empty’ particles in F192W mutant compared with WT. Scale bars, 100 nm. SDS–PAGE shows that the level of protein expression is comparable in F192W compared with WT in infiltrated leaves; however, F192W is unable to cause a systemic infection. (b) SDS–PAGE showing that the V109W mutation abolishes the assembly of eVLP. (c) SDS–PAGE showing R193D and E147R mutations also prevent eVLP assembly; however, the double mutation of E147R/R193D, which preserves the salt-bridge, permits very similar levels of capsid assembly compared with WT.
