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
Virologyjournal_title
Virologyauthors
Shin J,Phelan PJ,Chhum P,Bashkenova N,Yim S,Parker R,Gagnon D,Gjoerup O,Archambault J,Bullock PAdoi
10.1016/j.virol.2014.07.042subject
Has Abstractpub_date
2014-11-01 00:00:00pages
113-125eissn
0042-6822issn
1096-0341pii
S0042-6822(14)00357-2journal_volume
468-470pub_type
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