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  • Title: Increased frequency of specific locus mutation following human cytomegalovirus infection.
    Author: Albrecht T, Fons MP, Deng CZ, Boldogh I.
    Journal: Virology; 1997 Mar 31; 230(1):48-61. PubMed ID: 9126261.
    Abstract:
    The effect of human cytomegalovirus (HCMV) infection on the frequency of mutations at the hypoxanthine-guanine phosphoribosyl transferase (hprt) locus was studied in Chinese hamster lung V79 cells. When V79 cells were infected with HCMV (strain AD169) at multiplicities of 0.1 to 50 plaque forming units (PFU) per cell the presumptive mutation frequency, as determined by the number of 6-thioguanine-resistant (TGr) colonies, was increased up to 16.8-fold (P < 0.005), depending on the multiplicity of infection. Increases in the mutation frequency at the hprt locus were also observed for other laboratory-adapted HCMV strains (C-87, Davis) and for low passage clinical isolates (82-1, 84-2). The expression time required for the maximum increase in TGr colonies was 3 days and was consistent among the HCMV strains evaluated in this study. UV-irradiation of HCMV stock up to a dose of 9.6 x 10(4) ergs/mm2 increased the mutation frequency, but further exposure to UV light or to heat (56 degrees for 30 min) significantly decreased the frequency of TGr-resistant colonies, suggesting that expression of HCMV genes was involved in the mutation process. HCMV-induced TGr cells demonstrated substantially reduced (> 96%) incorporation of [3H]hypoxanthine. PCR analysis of the hprt locus demonstrated deletions in 9 of 19 HCMV-induced TGr colonies randomly selected for further study, while 2 of 17 spontaneously developed TGr colonies demonstrated deletions. Although insertions were not detected in spontaneously developed clones, 3 of 19 HCMV-induced TGr clones had insertions in the hprt gene. Neither HCMV-specific DNA sequences nor HCMV-specific proteins were detected in the TGr clones obtained after HCMV infection. Infection of V79 cells with HCMV also increased their sensitivity to mutation with N-methyl-N'-nitro-N-nitrosoguanidine, giving a synergistic enhancement of the mutation frequency. These results indicate that HCMV infection has the capacity to induce mutations in the cellular genome and increase the sensitivity of infected cells to mutation by genotoxic chemicals. Although inactivated HCMV particles are responsible for a modest increase in the mutation frequency, expression of HCMV genes is associated with a substantial enhancement of the mutation frequency.
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