Analysis of JC Virus DNA Replication Using a Quantitative and High-throughput Assay

Overview

Journal Virology

Specialty Microbiology

Date 2014 Aug 27

PMID 25155200

Citations 4

Authors

Jong Shin

Paul J Phelan

Panharith Chhum

Nazym Bashkenova

Sung Yim

Robert Parker

David Gagnon

Ole Gjoerup

Jacques Archambault

Peter A Bullock

Affiliations

Soon will be listed here.

Abstract

Progressive Multifocal Leukoencephalopathy (PML) is caused by lytic replication of JC virus (JCV) in specific cells of the central nervous system. Like other polyomaviruses, JCV encodes a large T-antigen helicase needed for replication of the viral DNA. Here, we report the development of a luciferase-based, quantitative and high-throughput assay of JCV DNA replication in C33A cells, which, unlike the glial cell lines Hs 683 and U87, accumulate high levels of nuclear T-ag needed for robust replication. Using this assay, we investigated the requirement for different domains of T-ag, and for specific sequences within and flanking the viral origin, in JCV DNA replication. Beyond providing validation of the assay, these studies revealed an important stimulatory role of the transcription factor NF1 in JCV DNA replication. Finally, we show that the assay can be used for inhibitor testing, highlighting its value for the identification of antiviral drugs targeting JCV DNA replication.

Citing Articles

DNA damage-induced phosphorylation of a replicative DNA helicase results in inhibition of DNA replication through attenuation of helicase function.

Homiski C, Dey-Rao R, Shen S, Qu J, Melendy T Nucleic Acids Res. 2024; 52(17):10311-10328.

PMID: 39126317 PMC: 11417368. DOI: 10.1093/nar/gkae663.

[Progress on pathogenesis of progressive multifocal leukoence-phalopathy].

Hu C, Zhu B Zhejiang Da Xue Xue Bao Yi Xue Ban. 2019; 47(5):534-540.

PMID: 30693697 PMC: 10393643. DOI: 10.3785/j.issn.1008-9292.2018.10.14.

Replication of JC Virus DNA in the G144 Oligodendrocyte Cell Line Is Dependent Upon Akt.

Peterson J, Lin B, Shin J, Phelan P, Tsichlis P, Schwob J J Virol. 2017; 91(20).

PMID: 28768870 PMC: 5625518. DOI: 10.1128/JVI.00735-17.

Structural Based Analyses of the JC Virus T-Antigen F258L Mutant Provides Evidence for DNA Dependent Conformational Changes in the C-Termini of Polyomavirus Origin Binding Domains.

Meinke G, Phelan P, Shin J, Gagnon D, Archambault J, Bohm A PLoS Pathog. 2016; 12(1):e1005362.

PMID: 26735515 PMC: 4703215. DOI: 10.1371/journal.ppat.1005362.

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