Identification of a Varicella-Zoster Virus Replication Inhibitor That Blocks Capsid Assembly by Interacting with the Floor Domain of the Major Capsid Protein

Inoue, Naoki; Matsushita, Mitsunori; Fukui, Yuichi; Yamada, Souichi; Tsuda, Mihoko; Higashi, Chizuka; Kaneko, Keiko; Hasegawa, Hideki; Yamaguchi, Toshiaki

doi:10.1128/jvi.01280-12

Cited by 23 publications

(23 citation statements)

References 61 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In contrast, MV9G cells have been used before to quantitate infection with two alphaherpesviruses (VZV and HSV-1) [22,25,59]. As demonstrated by these studies and the data presented here, MV9G cells enable comparative analyses of human herpesviruses of different herpesvirinae subfamilies in the same assay system and even as a virus mixture in the same culture which is another unique advantage over other available HCMV reporter cell lines.…”

Section: Discussionmentioning

confidence: 98%

“…Together, these data demonstrate that MV9G cells are suitable to study the effects of antiviral agents acting in different phases of the HCMV replication cycle. Furthermore, MV9G cells have been shown to be suitable for high-throughput screening of potential antiviral compounds against VZV [59] which might be useful to identify novel HCMV inhibitors in the future.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

A Luciferase Gene Driven by an Alphaherpesviral Promoter Also Responds to Immediate Early Antigens of the Betaherpesvirus HCMV, Allowing Comparative Analyses of Different Human Herpesviruses in One Reporter Cell Line

Maier¹,

Jung²,

Villinger

et al. 2017

PLoS ONE

View full text Add to dashboard Cite

Widely used methods for quantification of human cytomegalovirus (HCMV) infection in cell culture such as immunoblotting or plaque reduction assays are generally restricted to low throughput and require time-consuming evaluation. Up to now, only few HCMV reporter cell lines have been generated to overcome these restrictions and they are afflicted with other limitations because permanently expandable cell lines are normally not fully permissive to HCMV. In this work, a previously existing epithelial cell line hosting a luciferase gene under control of a Varicella-zoster virus promoter was adopted to investigate HCMV infection. The cells were susceptible to different HCMV strains at infection efficiencies that corresponded to their respective degree of epithelial cell tropism. Expression of early and late viral antigens, formation of nuclear inclusions, release of infectious virus progeny, and focal growth indicated productive viral replication. However, viral release and spread occurred at lower levels than in primary cell lines which appears to be due to a malfunction of virion morphogenesis during the nuclear stage. Expression of the luciferase reporter gene was specifically induced in HCMV infected cultures as a function of the virus dose and dependent on viral immediate early gene expression. The level of reporter activity accurately reflected infection efficiencies as determined by viral antigen immunostaining, and hence could discriminate the cell tropism of the tested virus strains. As proof-of-principle, we demonstrate that this cell line is applicable to evaluate drug resistance of clinical HCMV isolates and the neutralization capacity of human sera, and that it allows comparative and simultaneous analysis of HCMV and human herpes simplex virus type 1. In summary, the permanent epithelial reporter cell line allows robust, rapid and objective quantitation of HCMV infection and it will be particularly useful in higher throughput analyses as well as in comparative analyses of different human herpesviruses.

show abstract

Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

A Luciferase Gene Driven by an Alphaherpesviral Promoter Also Responds to Immediate Early Antigens of the Betaherpesvirus HCMV, Allowing Comparative Analyses of Different Human Herpesviruses in One Reporter Cell Line

Maier¹,

Jung²,

Villinger

et al. 2017

PLoS ONE

View full text Add to dashboard Cite

show abstract

“…V‐Oka‐infected HEL cells were cultured in the presence of DMSO, 20 µM 35B2, 20 µg/mL ACV and 40 µM 45B5. At 4 or 24 hr p.i., nuclei were prepared from the infected cells, and VZV copy numbers per cell were obtained by quantitative PCR assays as described previously . The relative copy numbers of viral DNA are shown using the copy numbers per cell obtained from the DMSO‐treated cells harvested at 24 hr p.i.…”

Section: Ec50 Of 45b5 Against Clinical Strains Acv‐sensitive and −Rementioning

confidence: 99%

“…The cells were infected with 100 PFU of cell‐free V‐Oka per well, and 2 hr later, culture media were replaced with media containing the same inhibitors. The cells were fixed 18 hr after the medium change and then stained with anti‐IE62 mAb in a manner similar to that described for the FRA . The relative number of IE62‐positive cells is shown using the number of IE62‐positive cells obtained from the DMSO‐treated cells as a 100% control.…”

Section: Ec50 Of 45b5 Against Clinical Strains Acv‐sensitive and −Rementioning

confidence: 99%

Characterization of an anti‐varicella‐zoster virus compound that targets the portal protein encoded by ORF54

Yasui

Yoshida

Yamaguchi

et al. 2017

Microbiology and Immunology

Self Cite

View full text Add to dashboard Cite

An anti-varicella-zoster virus compound, a 5-chlorobenzo[b]thiophen derivative (45B5), was characterized. Its 50% effective concentration against the cell-free vaccine Oka strain and 50% cytotoxic concentration in human fibroblasts were 16.9 µM and more than 100 µM, respectively. Treatment with 45B5 decreased viral DNA synthesis and IE62 expression weakly but significantly. All 45B5-resistant viral clones isolated were found to have at least one mutation in ORF54 that encodes the portal protein. There were no effects on interaction between the portal and scaffold proteins. Thus, 45B5 may inhibit nuclear delivery of viral DNA.

show abstract

“…Novel small molecules that target VZV capsid formation [16] and the HSV helicase-primase [17-19] are under study. Further investigation of herpesvirus portals and their associated terminase proteins will likely yield new targets for antiviral drug development.…”

mentioning

confidence: 99%

Non-Axial View of the Varicella-Zoster Virus Portal Protein Reveals Conserved Crown, Wing and Clip Architecture

Visalli

Howard

2014

Intervirology

View full text Add to dashboard Cite

Background: Herpesviridae encode a family of protein homologues that function as the ‘port of entry' for insertion of the viral DNA into preformed capsids during encapsidation. Methods: Transmission electron microscopy (TEM) of recombinant varicella-zoster virus pORF54 was performed. Results: Results suggest that pORF54 forms higher-order structures with itself. Enriched fractions analyzed by TEM revealed non-axial oriented portals with defined central channels and distinguishable crown, wing and clip regions. Conclusion: These morphological features are consistent with those previously reported for other herpesvirus and bacteriophage portal proteins.

show abstract

Identification of a Varicella-Zoster Virus Replication Inhibitor That Blocks Capsid Assembly by Interacting with the Floor Domain of the Major Capsid Protein

Cited by 23 publications

References 61 publications

A Luciferase Gene Driven by an Alphaherpesviral Promoter Also Responds to Immediate Early Antigens of the Betaherpesvirus HCMV, Allowing Comparative Analyses of Different Human Herpesviruses in One Reporter Cell Line

A Luciferase Gene Driven by an Alphaherpesviral Promoter Also Responds to Immediate Early Antigens of the Betaherpesvirus HCMV, Allowing Comparative Analyses of Different Human Herpesviruses in One Reporter Cell Line

Characterization of an anti‐varicella‐zoster virus compound that targets the portal protein encoded by ORF54

Non-Axial View of the Varicella-Zoster Virus Portal Protein Reveals Conserved Crown, Wing and Clip Architecture

Contact Info

Product

Resources

About