Structure of the Vesicular Stomatitis Virus N0-P Complex
Figure 2
Heterodimeric VSV NΔ210-P60 complex in solution.
(A) The absence of RNA in the complex is revealed by the UV absorbance spectrum, which exhibits an A280 nm/A260 nm ratio of 2.0. (B) Analysis of the NΔ210-P60 complex by SEC-MALLS. The complex elutes as a single peak at 15.6 mL (line), and the presence of the two proteins in the complex is demonstrated by SDS-PAGE analysis of the peak fraction using Coomassie blue staining (inset). The molecular mass of 53±2 kDa (crosses) indicates a 1∶1 complex between NΔ21 and P60 (calculated molecular mass = 45,377 Da (NΔ21)+8,053 Da (P60) = 53,430 Da). (C) Experimental SAXS data (open circles) up to 3.5 nm−1. The SAXS curve recorded at ESRF beamline ID 14-3 shows the scattering intensity I(q) as a function of the scattering vector, . (D) Average ab initio bead model of the NΔ210-P60 complex. The N protomer extracted from the circular N-RNA complex (2GIC chain E) fits to the SAXS-derived model, except for the NCT-loop.