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, 81 (21), 12077-9

Herpes Simplex Virus Mutants With Multiple Substitutions Affecting DNA Binding of UL42 Are Impaired for Viral Replication and DNA Synthesis

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Herpes Simplex Virus Mutants With Multiple Substitutions Affecting DNA Binding of UL42 Are Impaired for Viral Replication and DNA Synthesis

Changying Jiang et al. J Virol.

Abstract

Herpes simplex virus mutants with single substitutions that decrease DNA binding by the DNA polymerase processivity subunit UL42 are only modestly impaired for viral replication. In this study, recombinant viruses harboring two or four of these mutations were constructed. The more substitutions, the more severe the defects in viral replication and DNA synthesis, suggesting that DNA binding by UL42 is important for these processes.

Figures

FIG. 1.
FIG. 1.
UL42 mutants form smaller plaques on Vero cells than the wild type (wt). Plaques formed on Vero and V9 cells were analyzed as described in the text. Numbers shown below the plaques are average sizes (mm2) of at least 20 plaques ± standard deviations. *, the mutant with quadruple mutations formed only tiny plaques, which could be observed clearly only under a high magnification (×200). The plaques formed by other viruses were observed under a magnification of ×100 at 3 days postinfection.
FIG. 2.
FIG. 2.
Viral DNA synthesized by UL42 recombinants. Vero cells (1 × 105) were inoculated with UL42 recombinants at a multiplicity of infection of 3. Infected cells were harvested at different times postinfection. DNA was isolated from aliquots of both infected cells and medium containing cell-free virus. Purified DNA was serially diluted and quantified by real-time PCR as described in reference to determine the relative copy numbers of viral DNA. Data shown are averages from two independent experiments. Since the independent isolates of each virus exhibited similar kinetics, only the results for one recombinant (recombinants A in Table 1) are shown for clarity. WT, wild type.

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