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. 2003 Aug;77(15):8593-5.
doi: 10.1128/jvi.77.15.8593-8595.2003.

E4orf3 is necessary for enhanced S-phase replication of cell cycle-restricted subgroup C adenoviruses

Robin N Shepard  1 , David A Ornelles
Affiliations

E4orf3 is necessary for enhanced S-phase replication of cell cycle-restricted subgroup C adenoviruses

Robin N Shepard et al. J Virol. 2003 Aug.

Abstract

E1B-55K-mutant or E4orf6-mutant adenoviruses replicate more effectively after infecting cells in S phase than after infecting cells in G(1). Enhanced S-phase replication of the E4orf6-mutant viruses requires the E4orf3 gene. This report demonstrates that the E4orf3 gene is also required for enhanced S-phase replication of the E1B-55K-mutant virus.

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Figures

FIG. 1.
FIG. 1.
The E1B-55K/E4orf3 double-mutant virus exhibits similar growth kinetics but is more defective for replication than the E1B-55K single-mutant virus. Asynchronously growing HeLa cells were infected with the viruses indicated in the legend at a multiplicity of 10 PFU per cell. The cells and culture medium were collected at 1, 6, 12, 18, 24, 48, and 96 h postinfection, the cells were lysed, and the virus yield was measured by plaque assay with 293 cells. The value plotted at 0 h postinfection is the amount of virus used to infect the cells. Data shown are from a representative experiment performed three times with similar results.
FIG. 2.
FIG. 2.
The E1B-55K/E4orf3 double-mutant virus shows comparable growth in S-phase-infected cells and G1-infected cells. HeLa cells were synchronized to G1 or S phase and infected with the indicated viruses at a multiplicity of 10 PFU per cell. The status of the relevant viral genes (E1B-55K or E4orf3) is indicated below the histogram. Virus yield at 48 h postinfection was measured by plaque assay with 293 cells. The results shown are averages for three to five independent infections in three independent experiments. Error bars indicate the standard errors of the means. The difference in virus yield between G1- and S-phase-infected cells was significant only for dl1520-infected cells (P = 0.016, two-tailed Student's t test).
FIG. 3.
FIG. 3.
A comparable fraction of cells infected in G1 or S phase with the E1B-55K/E4orf3 double-mutant virus contain progeny. HeLa cells were synchronized to either G1 or S phase and infected with the indicated viruses at a multiplicity of 10 PFU per cell. The status of the relevant viral genes (E1B-55K or E4orf3) is indicated below the histogram. Cells that were clearly infected were scored for the presence or absence of progeny virus particles. At least 100 cells per experiment were evaluated. Each value represents the average percent ± standard deviation from three or four independent experiments.
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