Subcellular localization and some biochemical properties of the flavivirus Kunjin nonstructural proteins NS2A and NS4A

Virology
Jason M MackenzieE G Westaway

Abstract

In a previous study on the replication of Kunjin virus using immunoelectron microscopy (E. G. Westaway, J. M. Mackenzie, M. T. Kenney, M. K. Jones, and A. A. Khromykh, 1997, J. Virol. 71, 6650-6661), NS1 and NS3 were found associated with double-stranded RNA (dsRNA) within vesicle packets (VP) in infected Vero cells, suggesting that these induced membrane structures may be the cytoplasmic sites of RNA replication. NS2B and NS3 (comprising the virus-encoded protease) were colocalized within distinct paracrystalline (PC) or convoluted membranes (CM), also induced in the cytoplasm, suggesting that these membranes are the sites of proteolytic cleavage. In this study we found by immunofluorescence (IF) that the small hydrophobic nonstructural proteins NS2A and NS4A were located in discrete foci in the cytoplasm of infected cells at both 16 and 24 h postinfection, partially coincident with dsRNA foci. In cryosections of infected cells at 24 h, NS2A was located by immunogold labeling primarily within VP, associated with labeled dsRNA. NS2A fused to glutathione S-transferase (GST) bound strongly to the 3' untranslated region of Kunjin RNA and also to the proposed replicase components NS3 and NS5 in cell lysates. NS4A was localized by i...Continue Reading

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Citations

Apr 17, 2008·Nature Reviews. Microbiology·Sven Miller, Jacomine Krijnse-Locker
Jul 1, 2008·Nature Reviews. Microbiology·Catherine L MurrayCharles M Rice
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Feb 15, 2013·Journal of Virology·Xuping XiePei-Yong Shi
Apr 22, 2011·Journal of Virology·Qing-Yin WangPei-Yong Shi
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Mar 9, 2000·Journal of Virology·A A KhromykhE G Westaway
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