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Dual topology of the processed hepatitis C virus protein NS4B is influenced by the NS5A protein
KTH, School of Biotechnology (BIO), Molecular Biotechnology.
2006 (English)In: Journal of General Virology, ISSN 0022-1317, E-ISSN 1465-2099, Vol. 87, 3263-3272 p.Article in journal (Refereed) Published
Abstract [en]

Among the least-known hepatitis C virus proteins is the non-structural protein 4B (NS4B). It localizes to the endoplasmic reticulum (ER) membrane and induces membrane changes, resulting in a membranous web that is reported to be the locale for virus replication. A model was presented previously for the topology of recombinant HCV NS4B of the 1 a genotype based on in vitro data. In this model, the N-terminal tail of a considerable fraction of the NS4B molecules was translocated into the ER lumen via a post-translational process, giving the protein a dual transmembrane topology. It is now reported that translocation of the N terminus also occurs for processed NS4B expressed in cells in the context of the polyprotein. In the presence of NS5A, however, a lower degree of translocation was observed, which may indicate that NS5A influences the topology of NS4B. In vitro expression studies of NS4B from all major genotypes demonstrated that translocation of the N terminus to the ER lumen is conserved across genotypes. This clearly suggests an important function for this feature. Furthermore, when disrupting a previously reported amphipathic helix (AH) in the N terminus of NS4B, translocation was inhibited. As a disrupted AH also abolished the ability of NS4B to rearrange membranes, these data indicate for the first time an association between translocation of the N terminus and membrane rearrangement. Finally, the present experiments also confirm the predicted location of the first luminal loop to be around aa 112.

Place, publisher, year, edition, pages
2006. Vol. 87, 3263-3272 p.
Keyword [en]
ENDOPLASMIC-RETICULUM MEMBRANE, HCV RNA REPLICATION, COMPLETE CODING SEQUENCE, LARGE ENVELOPE PROTEIN, B-VIRUS, TRANSMEMBRANE TOPOLOGY, NONSTRUCTURAL PROTEINS, CELL-CULTURE, SUBGENOMIC REPLICONS, PREDOMINANT GENOTYPE
National Category
Industrial Biotechnology
Identifiers
URN: urn:nbn:se:kth:diva-37622DOI: 10.1099/vir.0.82211-0ISI: 000241542100015Scopus ID: 2-s2.0-33750249579OAI: oai:DiVA.org:kth-37622DiVA: diva2:434796
Note
QC 20110816Available from: 2011-08-16 Created: 2011-08-15 Last updated: 2017-12-08Bibliographically approved

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