Home LiteratureArticle Details
PMID: 12692249 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Identification of the hepatitis C virus RNA replication complex in Huh-7 cells harboring subgenomic replicons.

Journal of virology ·Vol. 77 ·No. 9 ·2003-05-00 ·Pages 5487-92

Gosert R, Egger D, Lohmann V, Bartenschlager R, Blum HE, Bienz K, Moradpour D

Abstract

Formation of a membrane-associated replication complex, composed of viral proteins, replicating RNA, and altered cellular membranes, is a characteristic feature of plus-strand RNA viruses. Here, we demonstrate the presence of a specific membrane alteration, designated the membranous web, that contains hepatitis C virus (HCV) nonstructural proteins, as well as viral plus-strand RNA, in Huh-7 cells harboring autonomously replicating subgenomic HCV RNAs. Metabolic labeling with 5-bromouridine 5'-triphosphate in the presence of actinomycin D revealed that the membranous web is the site of viral RNA synthesis and therefore represents the replication complex of HCV.

MeSH Terms
Cell Membrane/metabolism,ultrastructure,virology Hepacivirus/genetics,metabolism,ultrastructure Humans RNA, Viral/biosynthesis,genetics,ultrastructure Replicon Tumor Cells, Cultured Viral Nonstructural Proteins/metabolism Virus Replication
Chemicals
NS-5 protein, hepatitis C virus NS3 protein, hepatitis C virus RNA, Viral Viral Nonstructural Proteins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Gosert Rainer
Department of Medicine II, University of Freiburg, Germany.
Egger Denise
Lohmann Volker
Bartenschlager Ralf
Blum Hubert E
Bienz Kurt
Moradpour Darius
References (32)
32 references, click to expand
  1. A cytopathic and a cell culture adapted hepatitis A virus strain differ in cell killing but not in intracellular membrane rearrangements.
    Virology. 2000 Jan 5;266(1):157-69 PMID: 10612670
  2. Semliki Forest virus mRNA capping enzyme requires association with anionic membrane phospholipids for activity.
    EMBO J. 1999 Jun 1;18(11):3164-72 PMID: 10357827
  3. Efficient initiation of HCV RNA replication in cell culture.
    Science. 2000 Dec 8;290(5498):1972-4 PMID: 11110665
  4. Characterization of cell lines carrying self-replicating hepatitis C virus RNAs.
    J Virol. 2001 Feb;75(3):1252-64 PMID: 11152498
  5. Mutations in hepatitis C virus RNAs conferring cell culture adaptation.
    J Virol. 2001 Feb;75(3):1437-49 PMID: 11152517
  6. Requirements for assembly of poliovirus replication complexes and negative-strand RNA synthesis.
    J Virol. 2001 Apr;75(8):3841-50 PMID: 11264373
  7. Non-structural proteins 2 and 3 interact to modify host cell membranes during the formation of the arterivirus replication complex.
    J Gen Virol. 2001 May;82(Pt 5):985-94 PMID: 11297673
  8. The hepatitis C virus nonstructural protein 4B is an integral endoplasmic reticulum membrane protein.
    Virology. 2001 May 25;284(1):70-81 PMID: 11352669
  9. Determinants for membrane association of the hepatitis C virus RNA-dependent RNA polymerase.
    J Biol Chem. 2001 Nov 23;276(47):44052-63 PMID: 11557752
  10. Selectable subgenomic and genome-length dicistronic RNAs derived from an infectious molecular clone of the HCV-N strain of hepatitis C virus replicate efficiently in cultured Huh7 cells.
    J Virol. 2002 Mar;76(6):2997-3006 PMID: 11861865
  11. An amino-terminal amphipathic alpha-helix mediates membrane association of the hepatitis C virus nonstructural protein 5A.
    J Biol Chem. 2002 Mar 8;277(10):8130-9 PMID: 11744739
  12. RNA replication of mouse hepatitis virus takes place at double-membrane vesicles.
    J Virol. 2002 Apr;76(8):3697-708 PMID: 11907209
  13. Persistent and transient replication of full-length hepatitis C virus genomes in cell culture.
    J Virol. 2002 Apr;76(8):4008-21 PMID: 11907240
  14. A positive-strand RNA virus replication complex parallels form and function of retrovirus capsids.
    Mol Cell. 2002 Mar;9(3):505-14 PMID: 11931759
  15. Expression of hepatitis C virus proteins induces distinct membrane alterations including a candidate viral replication complex.
    J Virol. 2002 Jun;76(12):5974-84 PMID: 12021330
  16. Visualization and functional analysis of RNA-dependent RNA polymerase lattices.
    Science. 2002 Jun 21;296(5576):2218-22 PMID: 12077417
  17. Hepatitis C: molecular virology and antiviral targets.
    Trends Mol Med. 2002 Oct;8(10):476-82 PMID: 12383770
  18. The hepatitis C virus RNA-dependent RNA polymerase membrane insertion sequence is a transmembrane segment.
    J Virol. 2002 Dec;76(24):13088-93 PMID: 12438637
  19. Kinetics and location of poliovirus macromolecular synthesis in correlation to virus-induced cytopathology.
    Virology. 1980 Jan 30;100(2):390-9 PMID: 6243433
  20. Experimental non-A, non-B hepatitis: four types of cytoplasmic alteration in hepatocytes of infected chimpanzees.
    Virchows Arch B Cell Pathol Incl Mol Pathol. 1980;33(3):233-43 PMID: 6110271
  21. Association of polioviral proteins of the P2 genomic region with the viral replication complex and virus-induced membrane synthesis as visualized by electron microscopic immunocytochemistry and autoradiography.
    Virology. 1987 Sep;160(1):220-6 PMID: 2820130
  22. Structural and functional characterization of the poliovirus replication complex.
    J Virol. 1992 May;66(5):2740-7 PMID: 1313898
  23. Hepatitis C virus antigen in hepatocytes: immunomorphologic detection and identification.
    Gastroenterology. 1992 Aug;103(2):622-9 PMID: 1378804
  24. Active complete in vitro replication of nodavirus RNA requires glycerophospholipid.
    Proc Natl Acad Sci U S A. 1992 Dec 1;89(23):11136-40 PMID: 1454791
  25. Immunocytochemistry and in situ hybridization in the electron microscope: combined application in the study of virus-infected cells.
    Histochem Cell Biol. 1995 May;103(5):325-38 PMID: 7641067
  26. In situ detection of hepatitis C virus--a critical appraisal.
    J Hepatol. 1996;24(2 Suppl):43-51 PMID: 8836888
  27. Formation of plant RNA virus replication complexes on membranes: role of an endoplasmic reticulum-targeted viral protein.
    EMBO J. 1997 Jul 1;16(13):4049-59 PMID: 9233814
  28. Continuous human cell lines inducibly expressing hepatitis C virus structural and nonstructural proteins.
    Hepatology. 1998 Jul;28(1):192-201 PMID: 9657112
  29. Open reading frame 1a-encoded subunits of the arterivirus replicase induce endoplasmic reticulum-derived double-membrane vesicles which carry the viral replication complex.
    J Virol. 1999 Mar;73(3):2016-26 PMID: 9971782
  30. Nascent flavivirus RNA colocalized in situ with double-stranded RNA in stable replication complexes.
    Virology. 1999 May 25;258(1):108-17 PMID: 10329573
  31. Replication of subgenomic hepatitis C virus RNAs in a hepatoma cell line.
    Science. 1999 Jul 2;285(5424):110-3 PMID: 10390360
  32. Subcellular localization, stability, and trans-cleavage competence of the hepatitis C virus NS3-NS4A complex expressed in tetracycline-regulated cell lines.
    J Virol. 2000 Mar;74(5):2293-304 PMID: 10666260
Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/jvi.77.9.5487-5492.2003
Published
2003-05-00
Pages
5487-92
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC153965
Subset
IM
External Links
PubMed source DOI: 10.1128/jvi.77.9.5487-5492.2003 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2