title: Tracking the virus-like particles ofMacrobrachium rosenbergiinodavirus in insect cells.
abstract: Macrobrachium rosenbergiinodavirus (MrNv) poses a major threat to the prawn industry. Currently, no effective vaccine and treatment are available to prevent the spread ofMrNv. Its infection mechanism and localisation in a host cell are also not well characterised. TheMrNv capsid protein (MrNvc) produced inEscherichia coliself-assembled into virus-like particles (VLPs) resembling the native virus. Thus, fluorescein labelledMrNvc VLPs were employed as a model to study the virus entry and localisation inSpodoptera frugiperda, Sf9 cells. Through fluorescence microscopy and sub-cellular fractionation, theMrNvc was shown to enter Sf9 cells, and eventually arrived at the nucleus. The presence ofMrNvc within the cytoplasm and nucleus of Sf9 cells was further confirmed by the Z-stack imaging. The presence of ammonium chloride (NH4Cl), genistein, methyl-β-cyclodextrin or chlorpromazine (CPZ) inhibited the entry ofMrNvc into Sf9 cells, but cytochalasin D did not inhibit this process. This suggests that the internalisation ofMrNvc VLPs is facilitated by caveolae- and clathrin-mediated endocytosis. The whole internalisation process ofMrNvc VLPs into a Sf9 cell was recorded with live cell imaging. We have also identified a potential nuclear localisation signal (NLS) ofMrNvc through deletion mutagenesis and verified by classical-NLS mapping. Overall, this study provides an insight into the journey ofMrNvc VLPs in insect cells.
authors.affiliation: Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia , Serdang , Selangor , Malaysia.
authors.forename: Ummi Fairuz
authors.initials: UF
authors.lastname: Hanapi
authors.affiliation: Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia , Serdang , Selangor , Malaysia.
authors.forename: Chean Yeah
authors.initials: CY
authors.lastname: Yong
authors.affiliation: Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia , Serdang , Selangor , Malaysia.
authors.forename: Zee Hong
authors.initials: ZH
authors.lastname: Goh
authors.affiliation: Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia; Institute of Bioscience, Universiti Putra Malaysia, Serdang, Selangor, Malaysia.
authors.forename: Noorjahan Banu
authors.initials: NB
authors.lastname: Alitheen
authors.affiliation: Xiamen University Malaysia , Sepang , Selangor , Malaysia.
authors.forename: Swee Keong
authors.initials: SK
authors.lastname: Yeap
authors.affiliation: Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia; Institute of Bioscience, Universiti Putra Malaysia, Serdang, Selangor, Malaysia.
authors.forename: Wen Siang
authors.initials: WS
authors.lastname: Tan
keywords: Endosome; Nodavirus; Nuclear translocation; Sub-cellular localisation; Virus-like particle