Subcellular localization of hepatitis E virus (HEV) replicase

Rehman, Shagufta ; Kapur, Neeraj ; Durgapal, Hemlata ; Panda, Subrat Kumar (2008) Subcellular localization of hepatitis E virus (HEV) replicase Virology, 370 (1). pp. 77-92. ISSN 0042-6822

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Hepatitis E virus (HEV) is a hepatotropic virus with a single sense-strand RNA genome of ˜7.2 kb in length. Details of the intracellular site of HEV replication can pave further understanding of HEV biology. In-frame fusion construct of functionally active replicase-enhanced green fluorescent protein (EGFP) gene was made in eukaryotic expression vector. The functionality of replicase-EGFP fusion protein was established by its ability to synthesize negative-strand viral RNA in vivo, by strand-specific anchored RT-PCR and molecular beacon binding. Subcellular co-localization was carried out using organelle specific fluorophores and by immuno-electron microscopy. Fluorescence Resonance Energy Transfer (FRET) demonstrated the interaction of this protein with the 3' end of HEV genome. The results show localization of replicase on the endoplasmic reticulum membranes. The protein regions responsible for membrane localization was predicted and identified by use of deletion mutants. Endoplasmic reticulum was identified as the site of replicase localization and possible site of replication.

Item Type:Article
Source:Copyright of this article belongs to Elsevier Science.
Keywords:Replicase-EGFP; Negative-sense RNA; Molecular Beacon; FRET; Endoplasmic Reticulum
ID Code:36399
Deposited On:05 Jul 2011 11:13
Last Modified:17 May 2016 19:22

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