This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Gosert, R.
Right arrow Articles by Moradpour, D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Gosert, R.
Right arrow Articles by Moradpour, D.

 Previous Article  |  Next Article 

Journal of Virology, May 2003, p. 5487-5492, Vol. 77, No. 9
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.9.5487-5492.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Identification of the Hepatitis C Virus RNA Replication Complex in Huh-7 Cells Harboring Subgenomic Replicons

Rainer Gosert,1,2 Denise Egger,2 Volker Lohmann,3 Ralf Bartenschlager,3 Hubert E. Blum,1 Kurt Bienz,2 and Darius Moradpour1*

Department of Medicine II, University of Freiburg, D-79106 Freiburg,1 Department of Molecular Virology, University of Heidelberg, D-69120 Heidelberg, Germany,3 Institute for Medical Microbiology, University of Basel, CH-4003 Basel, Switzerland2

Received 18 October 2002/ Accepted 5 February 2003


arrow
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.


arrow
TEXT
 
Formation of a membrane-associated replication complex composed of viral proteins, replicating RNA, and altered cellular membranes is a hallmark of all of the plus-strand RNA viruses investigated thus far (4, 11, 23, 27, 29, 32). This strategy may offer multiple potential advantages, including (i) compartmentalization and local concentration of viral products, (ii) physical support and organization of the RNA replication complex (20), (iii) tethering of the viral RNA during unwinding, (iv) provision of lipid constituents important for replication (1, 34), and (v) protection of the viral RNA from double-stranded RNA-mediated host defenses or RNA interference.

Hepatitis C virus (HCV) contains a plus-strand RNA genome of approximately 9,600 nucleotides that encodes a polyprotein precursor of about 3,000 amino acid residues (17, 21). We have recently shown that expression of the HCV polyprotein in tetracycline-regulated human osteosarcoma cell lines induces distinct membrane alterations. A prominent alteration, designated the membranous web, was found to contain all of the viral structural and nonstructural proteins and therefore was proposed to represent the HCV replication complex (9). The membranous web could be induced by NS4B alone and was very similar to the "sponge-like inclusions" previously observed by electron microscopy (EM) in the livers of HCV-infected chimpanzees (24).

The recent development of replicon systems for HCV has allowed, for the first time, the study of efficient and genuine HCV RNA replication in Huh-7 human hepatoma cells in vitro (6, 13, 19, 25). The aims of the present study were to locate the site of HCV RNA replication and to identify the HCV replication complex in Huh-7 cells harboring a prototype subgenomic HCV replicon.

First, Huh-7-derived clone 9-13, harboring a bicistronic subgenomic replicon (19), was investigated by immunofluorescence microscopy (IF) to determine the subcellular distribution of the viral nonstructural proteins. Fixation, permeabilization, and immunostaining of cells were performed as previously described (10). As shown in Fig. 1a, NS3 was found in the cytoplasm as brightly fluorescing dots and in a reticular staining pattern. Very similar staining patterns have consistently been observed with monoclonal antibodies (MAbs) directed against NS4A, NS4B, NS5A, and NS5B (data not shown). By double-labeling IF, HCV nonstructural proteins were found to colocalize in the dot-like structures (data not shown). This is consistent with our observations for tetracycline-regulated cell lines expressing the entire HCV polyprotein (22) and suggests the formation of a multiprotein complex.



View larger version (62K):
[in this window]
[in a new window]
  		FIG. 1. Localization of HCV nonstructural proteins and plus-strand RNA in Huh-7 cells harboring a subgenomic replicon. (a) Huh-7-derived 9-13 replicon cells were examined by IF with HCV NS3-specific MAb 1B6 (33). NS3 was located in dot-like structures and in a reticular staining pattern. (b) HCV plus-strand RNA was detected by FISH with a directly FITC-labeled riboprobe of negative polarity. HCV plus-strand RNA accumulated in dot-like structures. Bar, 10 µm. Naive Huh-7 cells stained in parallel showed no specific staining.

To determine the distribution of viral plus-strand RNA, fluorescence in situ hybridization (FISH) was performed. A fluorescein isothiocyanate (FITC)-labeled riboprobe of negative polarity was prepared by in vitro transcription from HindIII-restricted plasmid pFKrepneo/R2884G (18) using T7 RNA polymerase (Roche Molecular Biochemicals, Mannheim, Germany). Labeling, alkaline hydrolysis, and purification of the 6.6-kb riboprobe were performed as previously described (10). HCV-specific plus-strand RNA was detectable as bright dots distributed over the cytoplasm, with some accumulation in the perinuclear region (Fig. 1b). The distribution and size of the dots strongly resembled those of the dots identified by IF (compare Fig. 1a and b). As commonly observed with riboprobes, there was some nonspecific labeling of the nucleoli of 9-13 replicon cells (Fig. 1b) and naive Huh-7 cells (data not shown). However, no label was found in the cytoplasm of naive Huh-7 cells, proving the specificity of the FISH reaction. In conclusion, both HCV nonstructural proteins and RNA localized to distinct, dot-like structures that are newly formed in Huh-7 cells harboring autonomously replicating HCV RNAs.

To identify the ultrastructural equivalent of the dot-like structures harboring viral proteins and RNA, we investigated naive and replicon-harboring Huh-7 cells by EM. For EM, cells were fixed in 2.5% glutaraldehyde and 2% OsO4 and embedded in Poly/Bed 812 (Polysciences, Warrington, Pa.) in accordance with standard protocols. As shown in Fig. 2a, no membrane alterations were found in naive Huh-7 cells. In contrast, a membranous web composed of small vesicles embedded in a membrane matrix was found in 9-13 cells (Fig. 2b and c). This structure was similar to the membranous web previously identified in U-2 OS human osteosarcoma-derived cell lines inducibly expressing the HCV polyprotein (9). As in these cell lines, the membranous web was often found closely associated with the rough endoplasmic reticulum (ER). On the basis of this observation, together with earlier studies demonstrating the colocalization of individually expressed HCV proteins with membranes of the ER (7, 12, 28, 33) and recent data indicating that HCV RNA replication takes place in a compartment that sustains endoglycosidase H-sensitive glycosylation (14), we speculate that the membranous web is derived from membranes of the ER.



View larger version (175K):
[in this window]
[in a new window]
  		FIG. 2. Membrane alterations in Huh-7 cells harboring a subgenomic replicon. (a) A naive Huh-7 cell displays unaltered organelles. Bar, 1 µm. (b) Low-power overview showing the membranous web (arrows) in a 9-13 replicon cell. Bar, 1 µm. (c) Higher magnification of the membranous web (arrows). A second membrane alteration that closely resembles the contiguous vesicles found in HCV-inducible cell lines and that did not harbor HCV proteins (9) was less abundant in replicon cells (arrowhead). Bar, 500 nm. G, Golgi apparatus; M, mitochondria; N, nucleus.

Immuno-EM was performed to investigate the localization of HCV nonstructural proteins. For immuno-EM, cells were fixed and embedded in LRGold (London Resin Co., London, United Kingdom) at -20°C as previously described (3). Sections were incubated with MAbs, followed by incubation with gold-conjugated secondary antibodies (Amersham Pharmacia Biotech, Piscataway, N.J.) as previously described (2). As shown at low magnification in Fig. 3a, NS5A-specific label strongly accumulated on a membranous structure. In addition, some label was found to be associated with the ER. Simultaneously processed naive Huh-7 cells did not show label accumulation above the background (Fig. 3b). Thus, distribution of NS5A at the ultrastructural level corresponds very well to the strong dot-like and weak reticular staining pattern observed by IF. Higher magnification allowed us to unequivocally define the structure extensively labeled by MAbs against NS5A and NS3 as the membranous web (Fig. 3c and d). Identical findings were obtained with MAbs against NS4A, NS4B, and NS5B (data not shown). Thus, all HCV nonstructural proteins are associated with the membranous web in Huh-7 cells harboring subgenomic HCV replicons. This result is in perfect agreement with our previous observations using tetracycline-regulated cell lines (9).



View larger version (200K):
[in this window]
[in a new window]
  		FIG. 3. Immunogold detection of HCV nonstructural proteins on the membranous web in replicon cells. (a) 9-13 replicon cells were labeled with NS5A-specific MAb 11H (7; kindly provided by Jan Albert Hellings, Organon Teknika B.V., Boxtel, The Netherlands). Gold particles accumulated on a membranous structure (surrounded by arrowheads). In addition, some label was found on the ER (arrows). Bar, 500 nm. (b) Identically treated naive Huh-7 cells were virtually devoid of gold particles. Bar, 500 nm. (c and d) At higher magnification, 9-13 replicon cells labeled with NS5A-specific MAb 11H (c) and NS3-specific MAb 1B6 (d). Both antigens are found almost exclusively on the membranous web. Bar, 200 nm.

The replicon system also enabled us to localize the genuinely replicated viral RNA by EM-in situ hybridization. A digoxigenin-labeled HCV-specific riboprobe of negative polarity was prepared that was analogous to the probe used for FISH (see above). After hybridization, RNA was detected with a gold-conjugated anti-digoxigenin antibody (Aurion, Wageningen, The Netherlands). The results shown in Fig. 4a reveal that the bulk of the HCV-specific plus-strand RNA was associated with the membranous web. An adjacent area of the same cell showed only background labeling (Fig. 4b). In conclusion, the membranous web contains, besides all of the HCV nonstructural proteins, the autonomously replicated viral RNA.



View larger version (163K):
[in this window]
[in a new window]
  		FIG. 4. EM-in situ hybridization of Huh-7 cells harboring a subgenomic HCV replicon. (a) HCV plus-strand RNA in 9-13 replicon cells, detected with a digoxigenin-labeled riboprobe of negative polarity and a gold-conjugated anti-digoxigenin antibody, is associated with the membranous web. (b) An adjacent area of the same cell shows no label accumulation above the background. Bar, 200 nm.

The observation that all of the viral nonstructural proteins localized to the membranous web allowed us to select arbitrarily one of the viral nonstructural proteins, i.e., NS5A, as a marker for the membranous web in subsequent IF studies. The observation that viral proteins and plus-strand RNA localized to the membranous web validated this membrane alteration as a candidate HCV replication complex. To conclusively identify the replication complex, we determined the site of viral RNA-dependent RNA synthesis by metabolic labeling with 5-bromouridine 5'-triphosphate as previously described (11). By double-labeling confocal laser scanning microscopy, we determined the localization of newly synthesized viral RNA in relation to NS5A as a marker of the membranous web. Digital optical sections were taken with a confocal laser scanning microscope (model TCS4D; Leica Lasertechnik, Heidelberg, Germany). NS5A was located with the polyclonal antiserum WU144 (kindly provided by Charles M. Rice, The Rockefeller University, New York, N.Y.) and an FITC-conjugated secondary antibody (green in Fig. 5a), and bromouridine-labeled newly synthesized viral RNA was detected with a MAb against bromodeoxyuridine (Bio Cell Consulting, Reinach, Switzerland), followed by a Texas Red-conjugated secondary antibody (red in Fig. 5b). Both signals clearly colocalized to the same cytoplasmic dot-like structures (yellow in Fig. 5c). The finding that NS5A, a marker of the membranous web, colocalizes with newly synthesized viral RNA demonstrates that the membranous web represents the site of viral RNA synthesis and, therefore, the HCV replication complex.



View larger version (5K):
[in this window]
[in a new window]
  		FIG. 5. Detection of nascent HCV RNA in Huh-7 cells harboring a subgenomic replicon. 9-13 replicon cells were metabolically labeled with 5-bromouridine 5'-triphosphate in the presence of actinomycin D and then subjected to double-labeling confocal laser scanning microscopy. (a) Detection of NS5A with the polyclonal antiserum WU144 and an FITC-conjugated secondary antibody. (b) Detection of newly synthesized, bromouridine-labeled viral RNA with a MAb against bromodeoxyuridine and a Texas Red-conjugated secondary antibody. (c) The overlay demonstrates colocalization (yellow) of NS5A and nascent viral RNA. Each image covers an area of 37 by 37 µm. Negative controls included identically treated naive Huh-7 cells.

The similarity of the membranous web identified in the replicon cells to that previously found in tetracycline-regulated cell lines indicates that expression of HCV proteins is sufficient to induce formation of the membranous web that accommodates active RNA replication. This is consistent with results obtained with other plus-strand RNA viruses where single proteins expressed alone or in the context of the entire open reading frame induced membrane alterations very similar to membrane changes found during natural infection (10, 30, 31). In the replicon cells, the membranous web was found less frequently than the inducible cell lines. Also, additional membrane alterations found in the tetracycline-regulated cell lines, notably, loose vesicles and contiguous vesicles, were much less abundant in the replicon cells. This is likely due to the overall lower protein expression levels in the replicon cells and may more closely reflect the natural situation in HCV-infected human and chimpanzee hepatocytes (15, 16). In this context, it is important to note that intracellular organelles, particularly the ER and the Golgi complex, remained intact in 9-13 cells. Thus, characteristic membrane changes described in other virus systems as cytopathic effects (5, 8) were not observed. This is consistent with unaltered growth characteristics of these cells compared to those of naive Huh-7 cells (26).

In our initial ultrastructural studies of Huh-7 cells carrying a persistently replicating HCV replicon, we did not detect the membranous web (26). The most likely reasons for this are differences in the specimen preparation methods that influence the recognizability of this structure in EM. Moreover, the previous identification of the membranous web in inducible cell lines that express high levels of the HCV polyprotein facilitated the finding of this structure in cells with a replicon where expression levels of HCV proteins are lower and formation of the membranous web is less frequent. Accordingly, we have very recently also identified the membranous web in Huh-7 cells harboring genomic length HCV replicons (25) (R.G. et al., unpublished observations).

In conclusion, ultrastructural analyses, combined with metabolic labeling of nascent viral RNA, have allowed us to identify, for the first time, the HCV replication complex in Huh-7 cells harboring autonomously replicating subgenomic HCV RNAs. Future studies will be aimed at isolating and further characterizing this complex and at defining the viral and cellular processes involved in formation of the membranous web. Elucidation of these processes will, in all likelihood, allow the definition of novel targets for antiviral intervention against hepatitis C.


arrow
ACKNOWLEDGMENTS
 
We gratefully acknowledge Lukas Landmann, Institute of Anatomy, University of Basel, Basel, Switzerland, for access to the confocal laser scanning microscope and Jan Albert Hellings and Charles M. Rice for antibodies.

This work was supported by grant Mo 799/1-3 and SFB 490, Teilprojekt A2 from the Deutsche Forschungsgemeinschaft, grant 31-055397.98 from the Swiss National Science Foundation, grants QLK2-CT1999-00356 and QLK2-CT2002-01329 from the European Commission, grant 01 KI 9951 from the Bundesministerium für Bildung und Forschung, the Lucie Bolte Foundation, and the Jubiläumsstiftung der Schweizerischen Rentenanstalt/Swiss Life.


arrow
FOOTNOTES
 
* Corresponding author. Mailing address: Department of Medicine II, University of Freiburg, D-79106 Freiburg, Germany. Phone: 49 761 270 3510. Fax: 49 761 270 3610. E-mail: Darius.Moradpour{at}uni-freiburg.de. Back


arrow
REFERENCES
 
    1
  1. Ahola, T., A. Lampio, P. Auvinen, and L. Kääriäinen. 1999. Semliki Forest virus mRNA capping enzyme requires association with anionic membrane phospholipids for activity. EMBO J. 18:3164-3172.[CrossRef][Medline]
    2. 2
  2. Bienz, K., and D. Egger. 1995. Immunocytochemistry and in situ hybridization in the electron microscope: combined application in the study of virus-infected cells. Histochem. Cell Biol. 103:325-338.[CrossRef][Medline]
    3. 3
  3. Bienz, K., D. Egger, and L. Pasamontes. 1987. 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 160:220-226.[CrossRef][Medline]
    4. 4
  4. Bienz, K., D. Egger, T. Pfister, and M. Troxler. 1992. Structural and functional characterization of the poliovirus replication complex. J. Virol. 66:2740-2747.[Abstract/Free Full Text]
    5. 5
  5. Bienz, K., D. Egger, Y. Rasser, and W. Bossart. 1980. Kinetics and location of poliovirus macromolecular synthesis in correlation to virus-induced cytopathology. Virology 100:390-399.[CrossRef][Medline]
    6. 6
  6. Blight, K. J., A. A. Kolykhalov, and C. M. Rice. 2000. Efficient initiation of HCV RNA replication in cell culture. Science 290:1972-1974.[Abstract/Free Full Text]
    7. 7
  7. Brass, V., E. Bieck, R. Montserret, B. Wölk, J. A. Hellings, H. E. Blum, F. Penin, and D. Moradpour. 2002. An amino-terminal amphipathic alpha-helix mediates membrane association of the hepatitis C virus nonstructural protein 5A. J. Biol. Chem. 277:8130-8139.[Abstract/Free Full Text]
    8. 8
  8. Egger, D., R. Gosert, and K. Bienz. 2002. Role of cellular structures in viral RNA replication, p. 247-253. In B. Semler and E. Wimmer (ed.), Molecular biology of picornaviruses. ASM Press, Washington, D.C.
    9. 9
  9. Egger, D., B. Wölk, R. Gosert, L. Bianchi, H. E. Blum, D. Moradpour, and K. Bienz. 2002. Expression of hepatitis C virus proteins induces distinct membrane alterations including a candidate viral replication complex. J. Virol. 76:5974-5984.[Abstract/Free Full Text]
    10. 10
  10. Gosert, R., D. Egger, and K. Bienz. 2000. A cytopathic and a cell culture adapted hepatitis A virus strain differ in cell killing but not in intracellular membrane rearrangements. Virology 266:157-169.[CrossRef][Medline]
    11. 11
  11. Gosert, R., A. Kanjanahaluethai, D. Egger, K. Bienz, and S. C. Baker. 2002. RNA replication of mouse hepatitis virus takes place at double-membrane vesicles. J. Virol. 76:3697-3708.[Abstract/Free Full Text]
    12. 12
  12. Hügle, T., F. Fehrmann, E. Bieck, M. Kohara, H.-G. Kräusslich, C. M. Rice, H. E. Blum, and D. Moradpour. 2001. The hepatitis C virus nonstructural protein 4B is an integral endoplasmic reticulum membrane protein. Virology 284:70-81.[CrossRef][Medline]
    13. 13
  13. Ikeda, M., M. Yi, K. Li, and S. M. Lemon. 2002. 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. 76:2997-3006.[Abstract/Free Full Text]
    14. 14
  14. Ivashkina, N., B. Wölk, V. Lohmann, R. Bartenschlager, H. E. Blum, F. Penin, and D. Moradpour. 2002. The hepatitis C virus RNA-dependent RNA polymerase membrane insertion sequence is a transmembrane segment. J. Virol. 76:13088-13093.[Abstract/Free Full Text]
    15. 15
  15. Krawczynski, K., M. J. Beach, D. W. Bradley, G. Kuo, A. M. Di Bisceglie, M. Houghton, G. R. Reyes, J. P. Kim, Q.-L. Choo, and M. J. Alter. 1992. Hepatitis C virus antigen in hepatocytes: immunomorphologic detection and identification. Gastroenterology 103:622-629.[Medline]
    16. 16
  16. Lau, J. Y. N., K. Krawczynski, F. Negro, and R. P. González-Peralta. 1996. In situ detection of hepatitis C virus—a critical appraisal. J. Hepatol. 24(Suppl. 2):43-51.[CrossRef][Medline]
    17. 17
  17. Lindenbach, B. D., and C. M. Rice. 2001. Flaviviridae: the viruses and their replication, p. 991-1041. In D. M. Knipe and P. M. Howley (ed.), Fields virology, 4th ed., vol. 1. Lippincott-Raven Publishers, Philadelphia, Pa.
    18. 18
  18. Lohmann, V., F. Körner, A. Dobierzewska, and R. Bartenschlager. 2001. Mutations in hepatitis C virus RNAs conferring cell culture adaptation. J. Virol. 75:1437-1449.[Abstract/Free Full Text]
    19. 19
  19. Lohmann, V., F. Körner, J.-O. Koch, U. Herian, L. Theilmann, and R. Bartenschlager. 1999. Replication of subgenomic hepatitis C virus RNAs in a hepatoma cell line. Science 285:110-113.[Abstract/Free Full Text]
    20. 20
  20. Lyle, J. M., E. Bullitt, K. Bienz, and K. Kirkegaard. 2002. Visualization and functional analysis of RNA-dependent RNA polymerase lattices. Science 296:2218-2222.[Abstract/Free Full Text]
    21. 21
  21. Moradpour, D., V. Brass, R. Gosert, B. Wölk, and H. E. Blum. 2002. Hepatitis C: molecular virology and antiviral targets. Trends Mol. Med. 8:476-482.[CrossRef][Medline]
    22. 22
  22. Moradpour, D., P. Kary, C. M. Rice, and H. E. Blum. 1998. Continuous human cell lines inducibly expressing hepatitis C virus structural and nonstructural proteins. Hepatology 28:192-201.[CrossRef][Medline]
    23. 23
  23. Pedersen, K. W., Y. van der Meer, N. Roos, and E. J. Snijder. 1999. 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. 73:2016-2026.[Abstract/Free Full Text]
    24. 24
  24. Pfeifer, U., R. Thomssen, K. Legler, U. Böttcher, W. Gerlich, E. Weinmann, and O. Klinge. 1980. Experimental non-A, non-B hepatitis: four types of cytoplasmic alteration in hepatocytes of infected chimpanzees. Virchows Arch. B Cell Pathol. Incl. Mol. Pathol. 33:233-243.
    25. 25
  25. Pietschmann, T., V. Lohmann, A. Kaul, N. Krieger, G. Rinck, G. Rutter, D. Strand, and R. Bartenschlager. 2002. Persistent and transient replication of full-length hepatitis C virus genomes in cell culture. J. Virol. 76:4008-4021.[Abstract/Free Full Text]
    26. 26
  26. Pietschmann, T., V. Lohmann, G. Rutter, K. Kurpanek, and R. Bartenschlager. 2001. Characterization of cell lines carrying self-replicating hepatitis C virus RNAs. J. Virol. 75:1252-1264.[Abstract/Free Full Text]
    27. 27
  27. Schaad, M. C., P. E. Jensen, and J. C. Carrington. 1997. Formation of plant RNA virus replication complexes on membranes: role of an endoplasmic reticulum-targeted viral protein. EMBO J. 16:4049-4059.[CrossRef][Medline]
    28. 28
  28. Schmidt-Mende, J., E. Bieck, T. Hügle, F. Penin, C. M. Rice, H. E. Blum, and D. Moradpour. 2001. Determinants for membrane association of the hepatitis C virus RNA-dependent RNA polymerase. J. Biol. Chem. 276:44052-44063.[Abstract/Free Full Text]
    29. 29
  29. Schwartz, M., J. Chen, M. Janda, M. Sullivan, J. den Boon, and P. Ahlquist. 2002. A positive-strand RNA virus replication complex parallels form and function of retrovirus capsids. Mol. Cell 9:505-514.[CrossRef][Medline]
    30. 30
  30. Snijder, E. J., H. van Tol, N. Roos, and K. W. Pedersen. 2001. Non-structural proteins 2 and 3 interact to modify host cell membranes during the formation of the arterivirus replication complex. J. Gen. Virol. 82:985-994.[Abstract/Free Full Text]
    31. 31
  31. Teterina, N. L., D. Egger, K. Bienz, D. M. Brown, B. L. Semler, and E. Ehrenfeld. 2001. Requirements for assembly of poliovirus replication complexes and negative-strand RNA synthesis. J. Virol. 75:3841-3850.[Abstract/Free Full Text]
    32. 32
  32. Westaway, E. G., A. A. Khromykh, and J. M. Mackenzie. 1999. Nascent flavivirus RNA colocalized in situ with double-stranded RNA in stable replication complexes. Virology 258:108-117.[CrossRef][Medline]
    33. 33
  33. Wölk, B., D. Sansonno, H.-G. Kräusslich, F. Dammacco, C. M. Rice, H. E. Blum, and D. Moradpour. 2000. Subcellular localization, stability and trans-cleavage competence of the hepatitis C virus NS3-NS4A complex expressed in tetracycline-regulated cell lines. J. Virol. 74:2293-2304.[Abstract/Free Full Text]
    34. 34
  34. Wu, S.-X., P. Ahlquist, and P. Kaesberg. 1992. Active complete in vitro replication of nodavirus RNA requires glycerophospholipid. Proc. Natl. Acad. Sci. USA 89:11136-11140.[Abstract/Free Full Text]

Journal of Virology, May 2003, p. 5487-5492, Vol. 77, No. 9
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.9.5487-5492.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Brohm, C., Steinmann, E., Friesland, M., Lorenz, I. C., Patel, A., Penin, F., Bartenschlager, R., Pietschmann, T. (2009). Characterization of Determinants Important for Hepatitis C Virus p7 Function in Morphogenesis by Using trans-Complementation. J. Virol. 83: 11682-11693 [Abstract] [Full Text]  
  • Gouttenoire, J., Montserret, R., Kennel, A., Penin, F., Moradpour, D. (2009). An Amphipathic {alpha}-Helix at the C Terminus of Hepatitis C Virus Nonstructural Protein 4B Mediates Membrane Association. J. Virol. 83: 11378-11384 [Abstract] [Full Text]  
  • Taguwa, S., Kambara, H., Omori, H., Tani, H., Abe, T., Mori, Y., Suzuki, T., Yoshimori, T., Moriishi, K., Matsuura, Y. (2009). Cochaperone Activity of Human Butyrate-Induced Transcript 1 Facilitates Hepatitis C Virus Replication through an Hsp90-Dependent Pathway. J. Virol. 83: 10427-10436 [Abstract] [Full Text]  
  • Yu, X., Sainz, B. Jr., Uprichard, S. L. (2009). Development of a Cell-Based Hepatitis C Virus Infection Fluorescent Resonance Energy Transfer Assay for High-Throughput Antiviral Compound Screening. Antimicrob. Agents Chemother. 53: 4311-4319 [Abstract] [Full Text]  
  • Amako, Y., Sarkeshik, A., Hotta, H., Yates, J. III, Siddiqui, A. (2009). Role of Oxysterol Binding Protein in Hepatitis C Virus infection. J. Virol. 83: 9237-9246 [Abstract] [Full Text]  
  • Dreux, M., Gastaminza, P., Wieland, S. F., Chisari, F. V. (2009). The autophagy machinery is required to initiate hepatitis C virus replication. Proc. Natl. Acad. Sci. USA 106: 14046-14051 [Abstract] [Full Text]  
  • Liu, Z., Yang, F., Robotham, J. M., Tang, H. (2009). Critical Role of Cyclophilin A and Its Prolyl-Peptidyl Isomerase Activity in the Structure and Function of the Hepatitis C Virus Replication Complex. J. Virol. 83: 6554-6565 [Abstract] [Full Text]  
  • Gouttenoire, J., Castet, V., Montserret, R., Arora, N., Raussens, V., Ruysschaert, J.-M., Diesis, E., Blum, H. E., Penin, F., Moradpour, D. (2009). Identification of a Novel Determinant for Membrane Association in Hepatitis C Virus Nonstructural Protein 4B. J. Virol. 83: 6257-6268 [Abstract] [Full Text]  
  • Hara, H., Aizaki, H., Matsuda, M., Shinkai-Ouchi, F., Inoue, Y., Murakami, K., Shoji, I., Kawakami, H., Matsuura, Y., Lai, M. M. C., Miyamura, T., Wakita, T., Suzuki, T. (2009). Involvement of Creatine Kinase B in Hepatitis C Virus Genome Replication through Interaction with the Viral NS4A Protein. J. Virol. 83: 5137-5147 [Abstract] [Full Text]  
  • Roohvand, F., Maillard, P., Lavergne, J.-P., Boulant, S., Walic, M., Andreo, U., Goueslain, L., Helle, F., Mallet, A., McLauchlan, J., Budkowska, A. (2009). Initiation of Hepatitis C Virus Infection Requires the Dynamic Microtubule Network: ROLE OF THE VIRAL NUCLEOCAPSID PROTEIN. J. Biol. Chem. 284: 13778-13791 [Abstract] [Full Text]  
  • Berger, K. L., Cooper, J. D., Heaton, N. S., Yoon, R., Oakland, T. E., Jordan, T. X., Mateu, G., Grakoui, A., Randall, G. (2009). Roles for endocytic trafficking and phosphatidylinositol 4-kinase III alpha in hepatitis C virus replication. Proc. Natl. Acad. Sci. USA 106: 7577-7582 [Abstract] [Full Text]  
  • Ujino, S., Yamaguchi, S., Shimotohno, K., Takaku, H. (2009). Heat-shock Protein 90 Is Essential for Stabilization of the Hepatitis C Virus Nonstructural Protein NS3. J. Biol. Chem. 284: 6841-6846 [Abstract] [Full Text]  
  • Jones, D. M., Patel, A. H., Targett-Adams, P., McLauchlan, J. (2009). The Hepatitis C Virus NS4B Protein Can trans-Complement Viral RNA Replication and Modulates Production of Infectious Virus. J. Virol. 83: 2163-2177 [Abstract] [Full Text]  
  • Jennings, T. A., Mackintosh, S. G., Harrison, M. K., Sikora, D., Sikora, B., Dave, B., Tackett, A. J., Cameron, C. E., Raney, K. D. (2009). NS3 Helicase from the Hepatitis C Virus Can Function as a Monomer or Oligomer Depending on Enzyme and Substrate Concentrations. J. Biol. Chem. 284: 4806-4814 [Abstract] [Full Text]  
  • Vassilaki, N., Friebe, P., Meuleman, P., Kallis, S., Kaul, A., Paranhos-Baccala, G., Leroux-Roels, G., Mavromara, P., Bartenschlager, R. (2008). Role of the Hepatitis C Virus Core+1 Open Reading Frame and Core cis-Acting RNA Elements in Viral RNA Translation and Replication. J. Virol. 82: 11503-11515 [Abstract] [Full Text]  
  • Ji, J., Glaser, A., Wernli, M., Berke, J. M., Moradpour, D., Erb, P. (2008). Suppression of short interfering RNA-mediated gene silencing by the structural proteins of hepatitis C virus. J. Gen. Virol. 89: 2761-2766 [Abstract] [Full Text]  
  • Paredes, A. M., Blight, K. J. (2008). A Genetic Interaction between Hepatitis C Virus NS4B and NS3 Is Important for RNA Replication. J. Virol. 82: 10671-10683 [Abstract] [Full Text]  
  • Wolk, B., Buchele, B., Moradpour, D., Rice, C. M. (2008). A Dynamic View of Hepatitis C Virus Replication Complexes. J. Virol. 82: 10519-10531 [Abstract] [Full Text]  
  • Nielsen, S. U., Bassendine, M. F., Martin, C., Lowther, D., Purcell, P. J., King, B. J., Neely, D., Toms, G. L. (2008). Characterization of hepatitis C RNA-containing particles from human liver by density and size. J. Gen. Virol. 89: 2507-2517 [Abstract] [Full Text]  
  • Lai, C.-K., Jeng, K.-S., Machida, K., Lai, M. M. C. (2008). Association of Hepatitis C Virus Replication Complexes with Microtubules and Actin Filaments Is Dependent on the Interaction of NS3 and NS5A. J. Virol. 82: 8838-8848 [Abstract] [Full Text]  
  • Okamoto, K., Mori, Y., Komoda, Y., Okamoto, T., Okochi, M., Takeda, M., Suzuki, T., Moriishi, K., Matsuura, Y. (2008). Intramembrane Processing by Signal Peptide Peptidase Regulates the Membrane Localization of Hepatitis C Virus Core Protein and Viral Propagation. J. Virol. 82: 8349-8361 [Abstract] [Full Text]  
  • Ma, Y., Yates, J., Liang, Y., Lemon, S. M., Yi, M. (2008). NS3 Helicase Domains Involved in Infectious Intracellular Hepatitis C Virus Particle Assembly. J. Virol. 82: 7624-7639 [Abstract] [Full Text]  
  • Steinmann, E., Brohm, C., Kallis, S., Bartenschlager, R., Pietschmann, T. (2008). Efficient trans-Encapsidation of Hepatitis C Virus RNAs into Infectious Virus-Like Particles. J. Virol. 82: 7034-7046 [Abstract] [Full Text]  
  • Bellecave, P., Cazenave, C., Rumi, J., Staedel, C., Cosnefroy, O., Andreola, M.-L., Ventura, M., Tarrago-Litvak, L., Astier-Gin, T. (2008). Inhibition of Hepatitis C Virus (HCV) RNA Polymerase by DNA Aptamers: Mechanism of Inhibition of In Vitro RNA Synthesis and Effect on HCV-Infected Cells. Antimicrob. Agents Chemother. 52: 2097-2110 [Abstract] [Full Text]  
  • Yang, F., Robotham, J. M., Nelson, H. B., Irsigler, A., Kenworthy, R., Tang, H. (2008). Cyclophilin A Is an Essential Cofactor for Hepatitis C Virus Infection and the Principal Mediator of Cyclosporine Resistance In Vitro. J. Virol. 82: 5269-5278 [Abstract] [Full Text]  
  • Bechill, J., Chen, Z., Brewer, J. W., Baker, S. C. (2008). Coronavirus Infection Modulates the Unfolded Protein Response and Mediates Sustained Translational Repression. J. Virol. 82: 4492-4501 [Abstract] [Full Text]  
  • Okamoto, T., Omori, H., Kaname, Y., Abe, T., Nishimura, Y., Suzuki, T., Miyamura, T., Yoshimori, T., Moriishi, K., Matsuura, Y. (2008). A Single-Amino-Acid Mutation in Hepatitis C Virus NS5A Disrupting FKBP8 Interaction Impairs Viral Replication. J. Virol. 82: 3480-3489 [Abstract] [Full Text]  
  • Huang, P., Goff, D. A., Huang, Q., Martinez, A., Xu, X., Crowder, S., Issakani, S. D., Anderson, E., Sheng, N., Achacoso, P., Yen, A., Kinsella, T., Darwish, I. S., Kolluri, R., Hong, H., Qu, K., Stauffer, E., Goldstein, E., Singh, R., Payan, D. G., Lu, H. H. (2008). Discovery and Characterization of Substituted Diphenyl Heterocyclic Compounds as Potent and Selective Inhibitors of Hepatitis C Virus Replication. Antimicrob. Agents Chemother. 52: 1419-1429 [Abstract] [Full Text]  
  • Targett-Adams, P., Boulant, S., McLauchlan, J. (2008). Visualization of Double-Stranded RNA in Cells Supporting Hepatitis C Virus RNA Replication. J. Virol. 82: 2182-2195 [Abstract] [Full Text]  
  • Tellinghuisen, T. L., Foss, K. L., Treadaway, J. C., Rice, C. M. (2008). Identification of Residues Required for RNA Replication in Domains II and III of the Hepatitis C Virus NS5A Protein. J. Virol. 82: 1073-1083 [Abstract] [Full Text]  
  • Yao, H., Ye, J. (2008). Long Chain Acyl-CoA Synthetase 3-mediated Phosphatidylcholine Synthesis Is Required for Assembly of Very Low Density Lipoproteins in Human Hepatoma Huh7 Cells. J. Biol. Chem. 283: 849-854 [Abstract] [Full Text]  
  • Supekova, L., Supek, F., Lee, J., Chen, S., Gray, N., Pezacki, J. P., Schlapbach, A., Schultz, P. G. (2008). Identification of Human Kinases Involved in Hepatitis C Virus Replication by Small Interference RNA Library Screening. J. Biol. Chem. 283: 29-36 [Abstract] [Full Text]  
  • Kaul, A., Woerz, I., Meuleman, P., Leroux-Roels, G., Bartenschlager, R. (2007). Cell Culture Adaptation of Hepatitis C Virus and In Vivo Viability of an Adapted Variant. J. Virol. 81: 13168-13179 [Abstract] [Full Text]  
  • Tang, W., Lazaro, C. A., Campbell, J. S., Parks, W. T., Katze, M. G., Fausto, N. (2007). Responses of Nontransformed Human Hepatocytes to Conditional Expression of Full-Length Hepatitis C Virus Open Reading Frame. Am. J. Pathol. 171: 1831-1846 [Abstract] [Full Text]  
  • Isken, O., Baroth, M., Grassmann, C. W., Weinlich, S., Ostareck, D. H., Ostareck-Lederer, A., Behrens, S.-E. (2007). Nuclear factors are involved in hepatitis C virus RNA replication. RNA 13: 1675-1692 [Abstract] [Full Text]  
  • Tellinghuisen, T. L., Evans, M. J., von Hahn, T., You, S., Rice, C. M. (2007). Studying Hepatitis C Virus: Making the Best of a Bad Virus. J. Virol. 81: 8853-8867 [Full Text]  
  • Blight, K. J. (2007). Allelic Variation in the Hepatitis C Virus NS4B Protein Dramatically Influences RNA Replication. J. Virol. 81: 5724-5736 [Abstract] [Full Text]  
  • Binder, M., Quinkert, D., Bochkarova, O., Klein, R., Kezmic, N., Bartenschlager, R., Lohmann, V. (2007). Identification of Determinants Involved in Initiation of Hepatitis C Virus RNA Synthesis by Using Intergenotypic Replicase Chimeras. J. Virol. 81: 5270-5283 [Abstract] [Full Text]  
  • Schaller, T., Appel, N., Koutsoudakis, G., Kallis, S., Lohmann, V., Pietschmann, T., Bartenschlager, R. (2007). Analysis of Hepatitis C Virus Superinfection Exclusion by Using Novel Fluorochrome Gene-Tagged Viral Genomes. J. Virol. 81: 4591-4603 [Abstract] [Full Text]  
  • Stone, M., Jia, S., Heo, W. D., Meyer, T., Konan, K. V. (2007). Participation of Rab5, an Early Endosome Protein, in Hepatitis C Virus RNA Replication Machinery. J. Virol. 81: 4551-4563 [Abstract] [Full Text]  
  • Van Wynsberghe, P. M., Chen, H.-R., Ahlquist, P. (2007). Nodavirus RNA Replication Protein A Induces Membrane Association of Genomic RNA. J. Virol. 81: 4633-4644 [Abstract] [Full Text]  
  • Kim, C. S., Seol, S. K., Song, O.-K., Park, J. H., Jang, S. K. (2007). An RNA-Binding Protein, hnRNP A1, and a Scaffold Protein, Septin 6, Facilitate Hepatitis C Virus Replication. J. Virol. 81: 3852-3865 [Abstract] [Full Text]  
  • Huang, H., Sun, F., Owen, D. M., Li, W., Chen, Y., Gale, M. Jr., Ye, J. (2007). From the Cover: Hepatitis C virus production by human hepatocytes dependent on assembly and secretion of very low-density lipoproteins. Proc. Natl. Acad. Sci. USA 104: 5848-5853 [Abstract] [Full Text]  
  • Brass, V., Pal, Z., Sapay, N., Deleage, G., Blum, H. E., Penin, F., Moradpour, D. (2007). Conserved Determinants for Membrane Association of Nonstructural Protein 5A from Hepatitis C Virus and Related Viruses. J. Virol. 81: 2745-2757 [Abstract] [Full Text]  
  • Quintavalle, M., Sambucini, S., Summa, V., Orsatti, L., Talamo, F., De Francesco, R., Neddermann, P. (2007). Hepatitis C Virus NS5A Is a Direct Substrate of Casein Kinase I-{alpha}, a Cellular Kinase Identified by Inhibitor Affinity Chromatography Using Specific NS5A Hyperphosphorylation Inhibitors. J. Biol. Chem. 282: 5536-5544 [Abstract] [Full Text]  
  • Jones, D. M., Gretton, S. N., McLauchlan, J., Targett-Adams, P. (2007). Mobility analysis of an NS5A-GFP fusion protein in cells actively replicating hepatitis C virus subgenomic RNA. J. Gen. Virol. 88: 470-475 [Abstract] [Full Text]  
  • Dahari, H., Ribeiro, R. M., Rice, C. M., Perelson, A. S. (2007). Mathematical Modeling of Subgenomic Hepatitis C Virus Replication in Huh-7 Cells. J. Virol. 81: 750-760 [Abstract] [Full Text]  
  • Lee, H., Liu, Y., Mejia, E., Paul, A. V., Wimmer, E. (2006). The C-Terminal Hydrophobic Domain of Hepatitis C Virus RNA Polymerase NS5B Can Be Replaced with a Heterologous Domain of Poliovirus Protein 3A. J. Virol. 80: 11343-11354 [Abstract] [Full Text]  
  • Lundin, M., Lindstrom, H., Gronwall, C., Persson, M. A. A. (2006). Dual topology of the processed hepatitis C virus protein NS4B is influenced by the NS5A protein.. J. Gen. Virol. 87: 3263-3272 [Abstract] [Full Text]  
  • Noppornpanth, S., Lien, T. X., Poovorawan, Y., Smits, S. L., Osterhaus, A. D. M. E., Haagmans, B. L. (2006). Identification of a naturally occurring recombinant genotype 2/6 hepatitis C virus.. J. Virol. 80: 7569-7577 [Abstract] [Full Text]  
  • Nomura-Takigawa, Y., Nagano-Fujii, M., Deng, L., Kitazawa, S., Ishido, S., Sada, K., Hotta, H. (2006). Non-structural protein 4A of Hepatitis C virus accumulates on mitochondria and renders the cells prone to undergoing mitochondria-mediated apoptosis. J. Gen. Virol. 87: 1935-1945 [Abstract] [Full Text]  
  • Lin, K., Perni, R. B., Kwong, A. D., Lin, C. (2006). VX-950, a Novel Hepatitis C Virus (HCV) NS3-4A Protease Inhibitor, Exhibits Potent Antiviral Activities in HCV Replicon Cells.. Antimicrob. Agents Chemother. 50: 1813-1822 [Abstract] [Full Text]  
  • Shimakami, T., Honda, M., Kusakawa, T., Murata, T., Shimotohno, K., Kaneko, S., Murakami, S. (2006). Effect of Hepatitis C Virus (HCV) NS5B-Nucleolin Interaction on HCV Replication with HCV Subgenomic Replicon.. J. Virol. 80: 3332-3340 [Abstract] [Full Text]  
  • Rouille, Y., Helle, F., Delgrange, D., Roingeard, P., Voisset, C., Blanchard, E., Belouzard, S., McKeating, J., Patel, A. H., Maertens, G., Wakita, T., Wychowski, C., Dubuisson, J. (2006). Subcellular Localization of Hepatitis C Virus Structural Proteins in a Cell Culture System That Efficiently Replicates the Virus. J. Virol. 80: 2832-2841 [Abstract] [Full Text]  
  • Munakata, T., Nakamura, M., Liang, Y., Li, K., Lemon, S. M. (2005). Down-regulation of the retinoblastoma tumor suppressor by the hepatitis C virus NS5B RNA-dependent RNA polymerase. Proc. Natl. Acad. Sci. USA 102: 18159-18164 [Abstract] [Full Text]  
  • Sandrin, V., Boulanger, P., Penin, F., Granier, C., Cosset, F.-L., Bartosch, B. (2005). Assembly of functional hepatitis C virus glycoproteins on infectious pseudoparticles occurs intracellularly and requires concomitant incorporation of E1 and E2 glycoproteins. J. Gen. Virol. 86: 3189-3199 [Abstract] [Full Text]  
  • Targett-Adams, P., McLauchlan, J. (2005). Development and characterization of a transient-replication assay for the genotype 2a hepatitis C virus subgenomic replicon. J. Gen. Virol. 86: 3075-3080 [Abstract] [Full Text]  
  • Quinkert, D., Bartenschlager, R., Lohmann, V. (2005). Quantitative Analysis of the Hepatitis C Virus Replication Complex. J. Virol. 79: 13594-13605 [Abstract] [Full Text]  
  • Mannova, P., Beretta, L. (2005). Activation of the N-Ras-PI3K-Akt-mTOR Pathway by Hepatitis C Virus: Control of Cell Survival and Viral Replication. J. Virol. 79: 8742-8749 [Abstract] [Full Text]  
  • Gosert, R., Jendrsczok, W., Berke, J. M., Brass, V., Blum, H. E., Moradpour, D. (2005). Characterization of Nonstructural Protein Membrane Anchor Deletion Mutants Expressed in the Context of the Hepatitis C Virus Polyprotein. J. Virol. 79: 7911-7917 [Abstract] [Full Text]  
  • Wolk, B., Gremion, C., Ivashkina, N., Engler, O. B., Grabscheid, B., Bieck, E., Blum, H. E., Cerny, A., Moradpour, D. (2005). Stable human lymphoblastoid cell lines constitutively expressing hepatitis C virus proteins. J. Gen. Virol. 86: 1737-1746 [Abstract] [Full Text]  
  • Svitkin, Y. V., Pause, A., Lopez-Lastra, M., Perreault, S., Sonenberg, N. (2005). Complete Translation of the Hepatitis C Virus Genome In Vitro: Membranes Play a Critical Role in the Maturation of All Virus Proteins except for NS3. J. Virol. 79: 6868-6881 [Abstract] [Full Text]  
  • Wilson, J. A., Richardson, C. D. (2005). Hepatitis C Virus Replicons Escape RNA Interference Induced by a Short Interfering RNA Directed against the NS5b Coding Region. J. Virol. 79: 7050-7058 [Abstract] [Full Text]  
  • Ferreon, J. C., Ferreon, A. C. M., Li, K., Lemon, S. M. (2005). Molecular Determinants of TRIF Proteolysis Mediated by the Hepatitis C Virus NS3/4A Protease. J. Biol. Chem. 280: 20483-20492 [Abstract] [Full Text]  
  • Gretton, S. N., Taylor, A. I., McLauchlan, J. (2005). Mobility of the hepatitis C virus NS4B protein on the endoplasmic reticulum membrane and membrane-associated foci. J. Gen. Virol. 86: 1415-1421 [Abstract] [Full Text]  
  • Tackett, A. J., Chen, Y., Cameron, C. E., Raney, K. D. (2005). Multiple Full-length NS3 Molecules Are Required for Optimal Unwinding of Oligonucleotide DNA in Vitro. J. Biol. Chem. 280: 10797-10806 [Abstract] [Full Text]  
  • Appel, N., Pietschmann, T., Bartenschlager, R. (2005). Mutational Analysis of Hepatitis C Virus Nonstructural Protein 5A: Potential Role of Differential Phosphorylation in RNA Replication and Identification of a Genetically Flexible Domain. J. Virol. 79: 3187-3194 [Abstract] [Full Text]  
  • Li, K., Foy, E., Ferreon, J. C., Nakamura, M., Ferreon, A. C. M., Ikeda, M., Ray, S. C., Gale, M. Jr., Lemon, S. M. (2005). Immune evasion by hepatitis C virus NS3/4A protease-mediated cleavage of the Toll-like receptor 3 adaptor protein TRIF. Proc. Natl. Acad. Sci. USA 102: 2992-2997 [Abstract] [Full Text]  
  • Appel, N., Herian, U., Bartenschlager, R. (2005). Efficient Rescue of Hepatitis C Virus RNA Replication by trans-Complementation with Nonstructural Protein 5A. J. Virol. 79: 896-909 [Abstract] [Full Text]  
  • Friebe, P., Boudet, J., Simorre, J.-P., Bartenschlager, R. (2005). Kissing-Loop Interaction in the 3' End of the Hepatitis C Virus Genome Essential for RNA Replication. J. Virol. 79: 380-392 [Abstract] [Full Text]  
  • Bigger, C. B., Guerra, B., Brasky, K. M., Hubbard, G., Beard, M. R., Luxon, B. A., Lemon, S. M., Lanford, R. E. (2004). Intrahepatic Gene Expression during Chronic Hepatitis C Virus Infection in Chimpanzees. J. Virol. 78: 13779-13792 [Abstract] [Full Text]  
  • Moradpour, D., Brass, V., Bieck, E., Friebe, P., Gosert, R., Blum, H. E., Bartenschlager, R., Penin, F., Lohmann, V. (2004). Membrane Association of the RNA-Dependent RNA Polymerase Is Essential for Hepatitis C Virus RNA Replication. J. Virol. 78: 13278-13284 [Abstract] [Full Text]  
  • Tellinghuisen, T. L., Marcotrigiano, J., Gorbalenya, A. E., Rice, C. M. (2004). The NS5A Protein of Hepatitis C Virus Is a Zinc Metalloprotein. J. Biol. Chem. 279: 48576-48587 [Abstract] [Full Text]  
  • Einav, S., Elazar, M., Danieli, T., Glenn, J. S. (2004). A Nucleotide Binding Motif in Hepatitis C Virus (HCV) NS4B Mediates HCV RNA Replication. J. Virol. 78: 11288-11295 [Abstract] [Full Text]  
  • Elazar, M., Liu, P., Rice, C. M., Glenn, J. S. (2004). An N-Terminal Amphipathic Helix in Hepatitis C Virus (HCV) NS4B Mediates Membrane Association, Correct Localization of Replication Complex Proteins, and HCV RNA Replication. J. Virol. 78: 11393-11400 [Abstract] [Full Text]  
  • Penin, F., Brass, V., Appel, N., Ramboarina, S., Montserret, R., Ficheux, D., Blum, H. E., Bartenschlager, R., Moradpour, D. (2004). Structure and Function of the Membrane Anchor Domain of Hepatitis C Virus Nonstructural Protein 5A. J. Biol. Chem. 279: 40835-40843 [Abstract] [Full Text]  
  • Panaviene, Z., Panavas, T., Serva, S., Nagy, P. D. (2004). Purification of the Cucumber Necrosis Virus Replicase from Yeast Cells: Role of Coexpressed Viral RNA in Stimulation of Replicase Activity. J. Virol. 78: 8254-8263 [Abstract] [Full Text]  
  • Moradpour, D., Evans, M. J., Gosert, R., Yuan, Z., Blum, H. E., Goff, S. P., Lindenbach, B. D., Rice, C. M. (2004). Insertion of Green Fluorescent Protein into Nonstructural Protein 5A Allows Direct Visualization of Functional Hepatitis C Virus Replication Complexes. J. Virol. 78: 7400-7409 [Abstract] [Full Text]  
  • Ma, Y., Shimakami, T., Luo, H., Hayashi, N., Murakami, S. (2004). Mutational Analysis of Hepatitis C Virus NS5B in the Subgenomic Replicon Cell Culture. J. Biol. Chem. 279: 25474-25482 [Abstract] [Full Text]  
  • Gao, L., Aizaki, H., He, J.-W., Lai, M. M. C. (2004). Interactions between Viral Nonstructural Proteins and Host Protein hVAP-33 Mediate the Formation of Hepatitis C Virus RNA Replication Complex on Lipid Raft. J. Virol. 78: 3480-3488 [Abstract] [Full Text]  
  • Lee, K. J., Choi, J., Ou, J.-h., Lai, M. M. C. (2004). The C-Terminal Transmembrane Domain of Hepatitis C Virus (HCV) RNA Polymerase Is Essential for HCV Replication In Vivo. J. Virol. 78: 3797-3802 [Abstract] [Full Text]  
  • Shimakami, T., Hijikata, M., Luo, H., Ma, Y. Y., Kaneko, S., Shimotohno, K., Murakami, S. (2004). Effect of Interaction between Hepatitis C Virus NS5A and NS5B on Hepatitis C Virus RNA Replication with the Hepatitis C Virus Replicon. J. Virol. 78: 2738-2748 [Abstract] [Full Text]  
  • WARIS, G., SARKER, S., SIDDIQUI, A. (2004). Two-step affinity purification of the hepatitis C virus ribonucleoprotein complex. RNA 10: 321-329 [Abstract] [Full Text]  
  • Ali, S., Pellerin, C., Lamarre, D., Kukolj, G. (2004). Hepatitis C Virus Subgenomic Replicons in the Human Embryonic Kidney 293 Cell Line. J. Virol. 78: 491-501 [Abstract] [Full Text]  
  • Miyanari, Y., Hijikata, M., Yamaji, M., Hosaka, M., Takahashi, H., Shimotohno, K. (2003). Hepatitis C Virus Non-structural Proteins in the Probable Membranous Compartment Function in Viral Genome Replication. J. Biol. Chem. 278: 50301-50308 [Abstract] [Full Text]  
  • El-Hage, N., Luo, G. (2003). Replication of hepatitis C virus RNA occurs in a membrane-bound replication complex containing nonstructural viral proteins and RNA. J. Gen. Virol. 84: 2761-2769 [Abstract] [Full Text]  
  • Zhong, W., An, H., Barawkar, D., Hong, Z. (2003). Dinucleotide Analogues as Novel Inhibitors of RNA-Dependent RNA Polymerase of Hepatitis C Virus. Antimicrob. Agents Chemother. 47: 2674-2681 [Abstract] [Full Text]  

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Gosert, R.
Right arrow Articles by Moradpour, D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Gosert, R.
Right arrow Articles by Moradpour, D.

Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»