Figure 7: Residues involved in encapsidation of genomic RNA.
From: Mechanisms of assembly and genome packaging in an RNA virus revealed by high-resolution cryo-EM
(a) SDS–PAGE and negative stain EM showing the R17D mutation in eVLP has little effect on capsid assembly, however, R17E completely abolishes it. EM images of the WT eVLP show different stain penetration depending on the presence or absence of the C terminus, with stain failing to penetrate complexes with an intact C terminus. The preparation in the EM is completely RNA free. (b) SDS–PAGE and negative stain EM showing R17 mutants in the L subunit. R17D and R17E abolish capsid assembly. Some flexibility is, however, tolerated as R17W, R17G and R17K all behave similarly to WT virus. (c) SDS–PAGE and negative stain EM showing W190 mutants in the L subunit. W190F abolishes RNA binding and particle assembly. However, W190A and W190D are indistinguishable from WT. All scale bars, 100 nm.
