TY - JOUR

T1 - Hepatitis C virus alters the morphology and function of peroxisomes

AU - Martin de Fourchambault, Esther

AU - Callens, Nathalie

AU - Saliou, Jean Michel

AU - Fourcot, Marie

AU - Delos, Oceane

AU - Barois, Nicolas

AU - Thorel, Quentin

AU - Ramirez, Santseharay

AU - Bukh, Jens

AU - Cocquerel, Laurence

AU - Bertrand-Michel, Justine

AU - Marot, Guillemette

AU - Sebti, Yasmine

AU - Dubuisson, Jean

AU - Rouillé, Yves

N1 - Publisher Copyright: Copyright © 2023 Martin de Fourchambault, Callens, Saliou, Fourcot, Delos, Barois, Thorel, Ramirez, Bukh, Cocquerel, Bertrand-Michel, Marot, Sebti, Dubuisson and Rouillé.

PY - 2023

Y1 - 2023

N2 - Despite the introduction of effective treatments for hepatitis C in clinics, issues remain regarding the liver disease induced by chronic hepatitis C virus (HCV) infection. HCV is known to disturb the metabolism of infected cells, especially lipid metabolism and redox balance, but the mechanisms leading to HCV-induced pathogenesis are still poorly understood. In an APEX2-based proximity biotinylation screen, we identified ACBD5, a peroxisome membrane protein, as located in the vicinity of HCV replication complexes. Confocal microscopy confirmed the relocation of peroxisomes near HCV replication complexes and indicated that their morphology and number are altered in approximately 30% of infected Huh-7 cells. Peroxisomes are small versatile organelles involved among other functions in lipid metabolism and ROS regulation. To determine their importance in the HCV life cycle, we generated Huh-7 cells devoid of peroxisomes by inactivating the PEX5 and PEX3 genes using CRISPR/Cas9 and found that the absence of peroxisomes had no impact on replication kinetics or infectious titers of HCV strains JFH1 and DBN3a. The impact of HCV on peroxisomal functions was assessed using sub-genomic replicons. An increase of ROS was measured in peroxisomes of replicon-containing cells, correlated with a significant decrease of catalase activity with the DBN3a strain. In contrast, HCV replication had little to no impact on cytoplasmic and mitochondrial ROS, suggesting that the redox balance of peroxisomes is specifically impaired in cells replicating HCV. Our study provides evidence that peroxisome function and morphology are altered in HCV-infected cells.

AB - Despite the introduction of effective treatments for hepatitis C in clinics, issues remain regarding the liver disease induced by chronic hepatitis C virus (HCV) infection. HCV is known to disturb the metabolism of infected cells, especially lipid metabolism and redox balance, but the mechanisms leading to HCV-induced pathogenesis are still poorly understood. In an APEX2-based proximity biotinylation screen, we identified ACBD5, a peroxisome membrane protein, as located in the vicinity of HCV replication complexes. Confocal microscopy confirmed the relocation of peroxisomes near HCV replication complexes and indicated that their morphology and number are altered in approximately 30% of infected Huh-7 cells. Peroxisomes are small versatile organelles involved among other functions in lipid metabolism and ROS regulation. To determine their importance in the HCV life cycle, we generated Huh-7 cells devoid of peroxisomes by inactivating the PEX5 and PEX3 genes using CRISPR/Cas9 and found that the absence of peroxisomes had no impact on replication kinetics or infectious titers of HCV strains JFH1 and DBN3a. The impact of HCV on peroxisomal functions was assessed using sub-genomic replicons. An increase of ROS was measured in peroxisomes of replicon-containing cells, correlated with a significant decrease of catalase activity with the DBN3a strain. In contrast, HCV replication had little to no impact on cytoplasmic and mitochondrial ROS, suggesting that the redox balance of peroxisomes is specifically impaired in cells replicating HCV. Our study provides evidence that peroxisome function and morphology are altered in HCV-infected cells.

KW - APEX2

KW - CRISPR-Cas9

KW - HCV genotype

KW - hepatitis C virus

KW - peroxisome

KW - proximity biotinylation

KW - ROS

U2 - 10.3389/fmicb.2023.1254728

DO - 10.3389/fmicb.2023.1254728

M3 - Journal article

C2 - 37808318

AN - SCOPUS:85173982518

VL - 14

JO - Frontiers in Microbiology

JF - Frontiers in Microbiology

SN - 1664-302X

M1 - 1254728

ER -