Inhibition of Human Cytomegalovirus DNA Maturation by a Benzimidazole Ribonucleoside Is Mediated through the UL89 Gene Product

Underwood, Mark; Harvey, Richard J.; Stanat, Sylvia C.; Hemphill, Mary Lou; Miller, Teresa L.; Drach, John C.; Townsend, Leroy B.; Biron, Karen K.

doi:10.1128/jvi.72.1.717-725.1998

Cited by 184 publications

(104 citation statements)

References 43 publications

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“…These assays were, however, compared for use in human CMV infections by Underwood et al who used simultaneous quantitative hybridization assays and plaque reduction assays to measure antiviral activity against HCMV. In these studies, an IC 50 in plaque reduction corresponded to an eightfold reduction in molecular viral load (26). Thus, the IC 90 by molecular viral load may be roughly comparable to the IC 50 using plaque reduction techniques.…”

Section: Discussionmentioning

confidence: 99%

Reduced Efficacy of Ganciclovir Against Porcine and Baboon Cytomegalovirus in Pig‐to‐Baboon Xenotransplantation

Mueller

Sulling²,

Gollackner³

et al. 2003

American Journal of Transplantation

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In pig‐to‐baboon xenotransplantation, porcine cytomegalovirus (PCMV) causes viremia, consumptive coagulopathy, and tissue‐invasive disease. Baboon cytomegalovirus (BCMV) is associated with invasive disease in xenograft recipients. The efficacy of prophylaxis with intravenous ganciclovir (GCV) was studied for prevention of PCMV and BCMV infections in pig‐to baboon xenotransplantation. GCV prophylaxis did not alter the incidence of BCMV activation in recipients, but reduced the amount of virus in tissues (mean 8.38 × 102 vs. 3.24 × 105 copies/µg DNA without treatment) and prevented tissue‐invasive infections. PCMV viral loads were unaltered by GCV prophylaxis (8.36 × 108 copies/µg DNA without prophylaxis vs. 1.20 × 109 copies/µg DNA with prophylaxis). In vitro, PCMV was relatively resistant to GCV [90% inhibitory concentration (IC90) of 10 µm, IC50 = 3 µm], acyclovir (100 µm), and leflunomide (not achievable). Only cidofovir (IC90 1 µm) and foscarnet (IC90 100 µm) might have therapeutic efficacy for PCMV in vivo in achievable concentrations, although these agents often carry significant toxicity in transplant recipients. GCV has limited activity against BCMV and no therapeutic efficacy against PCMV at standard doses in vivo. GCV and other antiviral agents have limited activities against PCMV in vitro. Breeding of PCMV‐free xenograft donors may be necessary to prevent PCMV infections in clinical trials.

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Section: Discussionmentioning

confidence: 99%

Reduced Efficacy of Ganciclovir Against Porcine and Baboon Cytomegalovirus in Pig‐to‐Baboon Xenotransplantation

Mueller

Sulling²,

Gollackner³

et al. 2003

American Journal of Transplantation

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“…Treatment of HCMV infected cells with either resulted in the accumulation of immature capsids and uncleaved DNA (Krosky et al, 1998). Maximal resistance to these compounds was mapped to two different encapsidation genes, UL89 and UL56 (Krosky et al, 1998;Underwood et al, 1998). The UL89/56 gene products are thought to make up the heterodimeric HCMV terminase complex.…”

Section: Discussionmentioning

confidence: 99%

The Varicella-zoster virus DNA encapsidation genes: Identification and characterization of the putative terminase subunits

et al. 2007

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The putative DNA encapsidation genes encoded by open reading frames (ORFs) 25, 26, 30, 34, 43, 45/42 and 54 were cloned from Varicella-zoster virus (VZV) strain Ellen. Sequencing revealed that the Ellen ORFs were highly conserved at the amino acid level when compared to those of 19 previously published VZV isolates. Additionally, RT-PCR provided the first evidence that ORF45/42 was expressed as a spliced transcript in VZV-infected cells. All seven ORFs were expressed in vitro and full length products were identified using a C-terminal V5 epitope tag. The in vitro products of the putative VZV terminase subunits encoded by ORFs 30 and 45/42 proved useful in protein-protein interaction assays. Previous studies have reported the formation of a heterodimeric terminase complex involved in DNA encapsidation for both herpes simplex virus-type 1 (HSV-1) and human cytomegalovirus (HCMV). Here we report that the C-terminal portion of exon II of ORF45/42 (ORF42-C269) interacted in GST-pull down experiments with in vitro synthesized ORF30 and ORF45/42. The interactions were maintained in the presence of anionic detergents and in buffers of increasing ionic strength. Cells transiently transfected with epitope tagged ORF45/42 or ORF30 showed primarily cytoplasmic staining. In contrast, an antiserum directed to the N-terminal portion of ORF45 showed nearly exclusive nuclear localization of the ORF45/42 gene product in infected cells. An ORF30 specific antiserum detected an 87 kDa protein in both the cytoplasmic and nuclear fractions of VZV infected cells. The results were consistent with the localization and function of herpesviral terminase subunits. This is the first study aimed at the identification and characterization of the VZV DNA encapsidation gene products.

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“…The mechanism by which BAY 38-4766 inhibits DNA cleavage and packaging appears to be quite similar to that of another class of maturational inhibitors, the benzimidazole ribosides, which include 2-bromo-5,6-dichloro-1-β-D-riborfuranosyl benzimidazole riboside (BDCRB) and 2,5,6-trichloro-1-β-D-riborfuranosyl benzimidazole riboside (TCRB; Krosky et al, 1998;Underwood et al, 1998). Despite pronounced dissimilarities in structure, amino acid changes that confer resistance to these compounds lie very close together in both terminase subunits (UL56 and UL89 in HCMV,and M56 and M89 in MCMV).…”

Section: Discussionmentioning

confidence: 98%

“…(B) Alignment of terminase amino acid sequences from GPCMV GP89 (residues 335-364), MCMV M89 (residues 338-367), and HCMV UL89 (residues 337-366). Mutations associated with resistance are color-coded: red, BDCRB/TCRBresistance in HCMV (Krosky et al, 1998;Underwood et al, 1998); purple, BAY 38-4766resistance in HCMV ; green, BAY 38-4766-resistance in MCMV Table 1 GPCMV dissemination in cyclophosphamide-treated strain 2 guinea pigs: impact of BAY 38-4766 on viral load on day 10 post-GPCMV challenge…”

Section: Figmentioning

confidence: 99%

The non-nucleoside antiviral, BAY 38-4766, protects against cytomegalovirus (CMV) disease and mortality in immunocompromised guinea pigs

et al. 2005

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New antiviral drugs are needed for the treatment of cytomegalovirus (CMV) infections, particularly in immunocompromised patients. These studies evaluated the in vitro and in vivo activity of the non-nucleosidic CMV inhibitor, BAY 38-4766, against guinea pig cytomegalovirus (GPCMV). Plaque reduction assays indicated that BAY 38-4766 was active against GPCMV, with an IC(50) of 0.5muM. Yield reduction assays demonstrated an ED(90) and ED(99) of 0.4 and 0.6muM, respectively, of BAY 38-4766 against GPCMV. Guinea pigs tolerated oral administration of 50mg/kg/day of BAY 38-4766 without evidence of biochemical or hematologic toxicity. Plasma concentrations of BAY 38-4766 were high following oral dosing, with a mean peak level at 1-h post-dose of 26.7mg/ml (n=6; range, 17.8-35.4). Treatment with BAY 38-4766 reduced both viremia and DNAemia, as determined by a real-time PCR assay, following GPCMV infection of cyclophosphamide-immunosuppressed strain 2 guinea pigs (p<0.05, Mann-Whitney test). BAY 38-4766 also reduced mortality following lethal GPCMV challenge in immunosuppressed Hartley guinea pigs, from 83% (20/24) in placebo-treated guinea pigs, to 17% (4/24) in BAY 38-4766-treated animals (p<0.0001, Fisher's exact test). Mortality differences were accompanied by reduction in DNAemia in Hartley guinea pigs. Based upon its favorable safety, pharmacokinetic, and therapeutic profiles, BAY 38-4766 warrants further investigation in the GPCMV model.

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Inhibition of Human Cytomegalovirus DNA Maturation by a Benzimidazole Ribonucleoside Is Mediated through the UL89 Gene Product

Cited by 184 publications

References 43 publications

Reduced Efficacy of Ganciclovir Against Porcine and Baboon Cytomegalovirus in Pig‐to‐Baboon Xenotransplantation

Reduced Efficacy of Ganciclovir Against Porcine and Baboon Cytomegalovirus in Pig‐to‐Baboon Xenotransplantation

The Varicella-zoster virus DNA encapsidation genes: Identification and characterization of the putative terminase subunits

The non-nucleoside antiviral, BAY 38-4766, protects against cytomegalovirus (CMV) disease and mortality in immunocompromised guinea pigs

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