Amphipathic α-Helices in Apolipoproteins Are Crucial to the Formation of Infectious Hepatitis C Virus Particles

PLoS Pathogens

Daisuke Okuzaki¹  Kazuhiko Koike²  Masahiro Yamamoto³  Shota Nakamura⁴  Toru Okamoto⁵  Masami Wada⁵  Takashi Motomura⁵  Takasuke Fukuhara⁵  Mai Shiokawa⁵  Chikako Ono⁵  Satomi Yamamoto⁵  Yoshiharu Matsuura⁵  Takaji Wakita⁶  Izumu Saito⁷ 

[1] DNA-Chip Developmental Center for Infectious Diseases, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan;Department of Gastroenterology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan;Department of Immunoparasitology, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan;Department of Infection Metagenomics, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan;Department of Molecular Virology, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan;Department of Virology II, National Institute of Infectious Diseases, Tokyo, Japan;Laboratory of Molecular Genetics, Institute of Medical Science, University of Tokyo, Tokyo, Japan

关键词: Apolipoproteins;    Vector-borne diseases;    Hepatitis C virus;    Small interfering RNAs;    Immunoblotting;    293T cells;    Apolipoprotein genes;    Focus-forming assay;   

DOI  :  10.1371/journal.ppat.1004534

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

Apolipoprotein B (ApoB) and ApoE have been shown to participate in the particle formation and the tissue tropism of hepatitis C virus (HCV), but their precise roles remain uncertain. Here we show that amphipathic α-helices in the apolipoproteins participate in the HCV particle formation by using zinc finger nucleases-mediated apolipoprotein B (ApoB) and/or ApoE gene knockout Huh7 cells. Although Huh7 cells deficient in either ApoB or ApoE gene exhibited slight reduction of particles formation, knockout of both ApoB and ApoE genes in Huh7 (DKO) cells severely impaired the formation of infectious HCV particles, suggesting that ApoB and ApoE have redundant roles in the formation of infectious HCV particles. cDNA microarray analyses revealed that ApoB and ApoE are dominantly expressed in Huh7 cells, in contrast to the high level expression of all of the exchangeable apolipoproteins, including ApoA1, ApoA2, ApoC1, ApoC2 and ApoC3 in human liver tissues. The exogenous expression of not only ApoE, but also other exchangeable apolipoproteins rescued the infectious particle formation of HCV in DKO cells. In addition, expression of these apolipoproteins facilitated the formation of infectious particles of genotype 1b and 3a chimeric viruses. Furthermore, expression of amphipathic α-helices in the exchangeable apolipoproteins facilitated the particle formation in DKO cells through an interaction with viral particles. These results suggest that amphipathic α-helices in the exchangeable apolipoproteins play crucial roles in the infectious particle formation of HCV and provide clues to the understanding of life cycle of HCV and the development of novel anti-HCV therapeutics targeting for viral assembly.

【 授权许可】

CC BY