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Comparative Study
. 2004 Sep;78(18):9837-41.
doi: 10.1128/JVI.78.18.9837-9841.2004.

Fitness analyses of vesicular stomatitis strains with rearranged genomes reveal replicative disadvantages

Isabel S Novella  1 , L Andrew Ball, Gail W Wertz
Affiliations
Comparative Study

Fitness analyses of vesicular stomatitis strains with rearranged genomes reveal replicative disadvantages

Isabel S Novella et al. J Virol. 2004 Sep.

Abstract

Gene expression of the nonsegmented negative-strand RNA viruses is determined by the position of each gene relative to that the single 3' promoter. The general order of genes among all of the viruses of the order Mononegavirales is highly conserved. In previous work we generated recombinant viruses in which the order of the three central genes of the prototypical rhabdovirus, vesicular stomatitis virus, was rearranged to all six possible permutations. While some of these viruses replicated less well than the wild type when assayed by single-step growth analyses in BSC-1 cells, others replicated as well or slightly better. In the work reported here, we used competition assays to compare the fitness of the viruses with alternative gene orders to that of the wild-type (wt) virus. We found that the relative fitness of these recombinant viruses depended on the multiplicity of infection (MOI) but not on the population size. However, during competitions at low MOI, when complementation cannot compensate for the defects of the populations with rearranged genomes, the virus with the wt gene order was always the most fit.

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Figures

FIG. 1.
FIG. 1.
Growth curves of recombinant VSV strains. Flasks were infected at an MOI of 0.001 to 0.005. Results show the averages of three independent experiments carried out with strains PMG (wt), GPM, and G1N2. Error bars represent the standard errors when they are larger than the symbol.
FIG. 2.
FIG. 2.
Fitness competitions between strains PMG (wt) and RU at an MOI of 0.001. The triplicate ratio measurements at each competition passage of a randomly chosen assay are presented. The solid line corresponds to the fit of the regression. The broken line indicates no changes in the relative ratios of strains PMG and RU.
FIG. 3.
FIG. 3.
Representative fitness assays of strains with rearranged genomes at an MOI of 0.001. Fitness assays are presented in two panels for clarity; note the difference in the scale of the y axes. Each point represents the average of the triplicate determination of ratios at each passage. Panel A shows competitions between strains RU and GMP (solid diamonds), RU and GPM (solid circles), and RU and G1N2 (open circles). Panel B shows competitions between strain RU and MGP (open squares) and RU and MPG (solid squares).
FIG. 4.
FIG. 4.
Fitness dependence on MOI during competition between strain RU and the rearranged strains. The MOI in these experiments was increased by increasing the total population size of the inoculum. Error bars show the standard error of each data point. The figure shows the results for strains GMP (solid diamonds), MGP (solid squares), GPM (solid triangles), MPG (solid circles), and G1N2 (open circles).
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