Detection of a fourth orbivirus non-structural protein.Reportar como inadecuado




Detection of a fourth orbivirus non-structural protein. - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

Reference: Belhouchet, M, Mohd Jaafar, F, Firth, AE et al., (2011). Detection of a fourth orbivirus non-structural protein. PloS one, 6 (10), Article: e25697.Citable link to this page:

 

Detection of a fourth orbivirus non-structural protein.

Abstract: The genus Orbivirus includes both insect and tick-borne viruses. The orbivirus genome, composed of 10 segments of dsRNA, encodes 7 structural proteins (VP1-VP7) and 3 non-structural proteins (NS1-NS3). An open reading frame (ORF) that spans almost the entire length of genome segment-9 (Seg-9) encodes VP6 (the viral helicase). However, bioinformatic analysis recently identified an overlapping ORF (ORFX) in Seg-9. We show that ORFX encodes a new non-structural protein, identified here as NS4. Western blotting and confocal fluorescence microscopy, using antibodies raised against recombinant NS4 from Bluetongue virus (BTV, which is insect-borne), or Great Island virus (GIV, which is tick-borne), demonstrate that these proteins are synthesised in BTV or GIV infected mammalian cells, respectively. BTV NS4 is also expressed in Culicoides insect cells. NS4 forms aggregates throughout the cytoplasm as well as in the nucleus, consistent with identification of nuclear localisation signals within the NS4 sequence. Bioinformatic analyses indicate that NS4 contains coiled-coils, is related to proteins that bind nucleic acids, or are associated with membranes and shows similarities to nucleolar protein UTP20 (a processome subunit). Recombinant NS4 of GIV protects dsRNA from degradation by endoribonucleases of the RNAse III family, indicating that it interacts with dsRNA. However, BTV NS4, which is only half the putative size of the GIV NS4, did not protect dsRNA from RNAse III cleavage. NS4 of both GIV and BTV protect DNA from degradation by DNAse. NS4 was found to associate with lipid droplets in cells infected with BTV or GIV or transfected with a plasmid expressing NS4.

Publication status:Published Funder: Department for Environment, Food and Rural Affairs   Funder: Biotechnology and Biological Sciences Research Council   Funder: Pfizer   Funder: European Union   Funder: Wellcome Trust   Notes:Copyright 2011 Belhouchet et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Bibliographic Details

Publisher: Public Library of Science

Publisher Website: http://www.plos.org

Journal: PloS onesee more from them

Publication Website: http://www.plosone.org

Issue Date: 2011

pages:Article: e25697Identifiers

Urn: uuid:8995a867-cdcf-4020-bf67-c082f81d4e26

Source identifier: 198972

Eissn: 1932-6203

Doi: https://doi.org/10.1371/journal.pone.0025697

Issn: 1932-6203 Item Description

Type: Journal article;

Language: eng Keywords: Animals Subcellular Fractions Cell Line Cell Nucleus Cell Nucleolus Orbivirus Reoviridae Infections Viral Nonstructural Proteins RNA, Double-Stranded Recombinant Proteins Microscopy, Fluorescence Cloning, Molecular Transfection Nuclease Protection Assays Colorimetry Blotting, Western Conserved Sequence Protein Binding Sequence Homology, Amino Acid Protein Transport Amino Acid Sequence Molecular Sequence Data Open Reading Frames Computational Biology Hydrophobic and Hydrophilic Interactions Tiny URL: pubs:198972

Relationships





Autor: Belhouchet, M - institutionUniversity of Oxford fundingBiotechnology and Biological Sciences Research Council fundingPfizer - - -

Fuente: https://ora.ox.ac.uk/objects/uuid:8995a867-cdcf-4020-bf67-c082f81d4e26



DESCARGAR PDF




Documentos relacionados