Identification of Interactions between Sindbis Virus Capsid Protein and Cytoplasmic vRNA as Novel Virulence Determinants
A) The RNA decay profiles of the incoming viral genomic RNAs of the individual SINV C:R interaction site mutants and parental wild type virus were determined by qRT-PCR analysis as described in the materials and methods. Plotted is the relative abundance of the incoming viral genomic RNAs (y-axis) with regards to time (x-axis). Regression analysis was utilized to determine the RNA decay profile (as shown with the solid line) and the dashed lines represent the 95% confidence intervals of the aforementioned regression. B) The half-lives of the individual genomic RNAs as determined using the calculations reported in Dolken et al., as determined by the first point at which the relative abundance has reached 0.5. C) The levels of Nanoluciferase activity for wild type parental virus and the nt10100 and nt10400 C:R interaction site mutants were determined as reported in the materials and methods at the indicated times post infection. All quantitative data in this figure represents the mean of at least three independent biological replicates. Comparative analysis was performed using variable bootstrapping, as described in the materials and methods, with the error bar representing the standard deviation of the mean. Statistical significance, as indicated on the individual panels above, are the p-Values obtained from Student’s t-test.