Publication: Development of viable TAP-tagged dengue virus for investigation of host–virus interactions in viral replication
Issued Date
2016-03-01
Resource Type
ISSN
14652099
00221317
00221317
Other identifier(s)
2-s2.0-84960465947
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Mahidol University
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SCOPUS
Bibliographic Citation
Journal of General Virology. Vol.97, No.3 (2016), 646-658
Suggested Citation
Teera Poyomtip, Kenneth Hodge, Ponpan Matangkasombut, Anavaj Sakuntabhai, Trairak Pisitkun, Siwanon Jirawatnotai, Sarin Chimnaronk Development of viable TAP-tagged dengue virus for investigation of host–virus interactions in viral replication. Journal of General Virology. Vol.97, No.3 (2016), 646-658. doi:10.1099/jgv.0.000371 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/40871
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Title
Development of viable TAP-tagged dengue virus for investigation of host–virus interactions in viral replication
Abstract
© 2016 The Authors. Dengue virus (DENV) is a mosquito-borne flavivirus responsible for life-threatening dengue haemorrhagic fever (DHF) and dengue shock syndrome (DSS). The viral replication machinery containing the core non-structural protein 5 (NS5) is implicated in severe dengue symptoms but molecular details remain obscure. To date, studies seeking to catalogue and characterize interaction networks between viral NS5 and host proteins have been limited to the yeast twohybrid system, computational prediction and co-immunoprecipitation (IP) of ectopically expressed NS5. However, these traditional approaches do not reproduce a natural course of infection in which a number of DENV NS proteins colocalize and tightly associate during the replication process. Here, we demonstrate the development of a recombinant DENV that harbours a TAP tag in NS5 to study host–virus interactions in vivo.We show that our engineered DENV was infective in several human cell lines and that the tags were stable over multiple viral passages, suggesting negligible structural and functional disturbance of NS5. We further provide proof-of-concept for the use of rationally tagged virus by revealing a high confidence NS5 interaction network in human hepatic cells. Our analysis uncovered previously unrecognized hnRNP complexes and several low-abundance fatty acid metabolism genes, which have been implicated in the viral life cycle. This study sets a new standard for investigation of host–flavivirus interactions.