Comparative Analysis of Hepatitis C Virus NS5A Dynamics and Localization in Assembly-Deficient Mutants

Pathogens

Costin-Ioan Popescu¹  Jean Dubuisson²  Yves Rouillé²  Laura Riva²  Nicolas Barois²  Corentin Spriet³ 

[1] Institute of Biochemistry of the Romanian Academy, 060031 Bucharest, Romania;University of Lille, CNRS, Inserm, Institut Pasteur de Lille, CHU Lille, U1019-UMR 8204-CIIL-Center for Infection and Immunity of Lille, 59000 Lille, France;University of Lille, CNRS, UMR 8576-UGSF-Department of Functional and Structural Glycobiology, 59000 Lille, France;

关键词: hepatitis C virus;    NS5A dynamics;    NS4B;    core;    lipid droplets;    assembly;   

DOI  :  10.3390/pathogens10020172

来源: DOAJ

【 摘 要 】

The hepatitis C virus (HCV) life cycle is a tightly regulated process, during which structural and non-structural proteins cooperate. However, the interplay between HCV proteins during genomic RNA replication and progeny virion assembly is not completely understood. Here, we studied the dynamics and intracellular localization of non-structural 5A protein (NS5A), which is a protein involved both in genome replication and encapsidation. An NS5A-eGFP (enhanced green fluorescent protein) tagged version of the strain JFH-1-derived wild-type HCV was compared to the corresponding assembly-deficient viruses ∆core, NS5A basic cluster 352–533 mutant (BCM), and serine cluster 451 + 454 + 457 mutant (SC). These analyses highlighted an increase of NS5A motility when the viral protein core was lacking. Although to a lesser extent, NS5A motility was also increased in the BCM virus, which is characterized by a lack of interaction of NS5A with the viral RNA, impairing HCV genome encapsidation. This observation suggests that the more static NS5A population is mainly involved in viral assembly rather than in RNA replication. Finally, NS4B exhibited a reduced co-localization with NS5A and lipid droplets for both ∆core and SC mutants, which is characterized by the absence of interaction of NS5A with core. This observation strongly suggests that NS5A is involved in targeting NS4B to lipid droplets (LDs). In summary, this work contributes to a better understanding of the interplay between HCV proteins during the viral life cycle.

【 授权许可】

Unknown