Analysis of JC virus DNA replication using a quantitative and high-throughput assay.

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.

journal_name

Virology

journal_title

Virology

authors

Shin J,Phelan PJ,Chhum P,Bashkenova N,Yim S,Parker R,Gagnon D,Gjoerup O,Archambault J,Bullock PA

doi

10.1016/j.virol.2014.07.042

subject

Has Abstract

pub_date

2014-11-01 00:00:00

pages

113-125

eissn

0042-6822

issn

1096-0341

pii

S0042-6822(14)00357-2

journal_volume

468-470

pub_type

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