Abstract:
UNLABELLED:Cell culture systems reproducing virus replication can serve as unique models for the discovery of novel bioactive molecules. Here, using a hepatitis C virus (HCV) cell culture system, we identified neoechinulin B (NeoB), a fungus-derived compound, as an inhibitor of the liver X receptor (LXR). NeoB was initially identified by chemical screening as a compound that impeded the production of infectious HCV. Genome-wide transcriptome analysis and reporter assays revealed that NeoB specifically inhibits LXR-mediated transcription. NeoB was also shown to interact directly with LXRs. Analysis of structural analogs suggested that the molecular interaction of NeoB with LXR correlated with the capacity to inactivate LXR-mediated transcription and to modulate lipid metabolism in hepatocytes. Our data strongly suggested that NeoB is a novel LXR antagonist. Analysis using NeoB as a bioprobe revealed that LXRs support HCV replication: LXR inactivation resulted in dispersion of double-membrane vesicles, putative viral replication sites. Indeed, cells treated with NeoB showed decreased replicative permissiveness for poliovirus, which also replicates in double-membrane vesicles, but not for dengue virus, which replicates via a distinct membrane compartment. Together, our data suggest that LXR-mediated transcription regulates the formation of virus-associated membrane compartments. Significantly, inhibition of LXRs by NeoB enhanced the activity of all known classes of anti-HCV agents, and NeoB showed especially strong synergy when combined with interferon or an HCV NS5A inhibitor. Thus, our chemical genetics analysis demonstrates the utility of the HCV cell culture system for identifying novel bioactive molecules and characterizing the virus-host interaction machinery. IMPORTANCE:Hepatitis C virus (HCV) is highly dependent on host factors for efficient replication. In the present study, we used an HCV cell culture system to screen an uncharacterized chemical library. Our results identified neoechinulin B (NeoB) as a novel inhibitor of the liver X receptor (LXR). NeoB inhibited the induction of LXR-regulated genes and altered lipid metabolism. Intriguingly, our results indicated that LXRs are critical to the process of HCV replication: LXR inactivation by NeoB disrupted double-membrane vesicles, putative sites of viral replication. Moreover, NeoB augmented the antiviral activity of all known classes of currently approved anti-HCV agents without increasing cytotoxicity. Thus, our strategy directly links the identification of novel bioactive compounds to basic virology and the development of new antiviral agents.
journal_name
J Viroljournal_title
Journal of virologyauthors
Nakajima S,Watashi K,Ohashi H,Kamisuki S,Izaguirre-Carbonell J,Kwon AT,Suzuki H,Kataoka M,Tsukuda S,Okada M,Moi ML,Takeuchi T,Arita M,Suzuki R,Aizaki H,Kato T,Suzuki T,Hasegawa H,Takasaki T,Sugawara F,Wakita Tdoi
10.1128/JVI.00856-16subject
Has Abstractpub_date
2016-09-29 00:00:00pages
9058-74issue
20eissn
0022-538Xissn
1098-5514pii
JVI.00856-16journal_volume
90pub_type
杂志文章abstract::The NS3 protein of hepatitis C virus (HCV) possesses protease activity responsible for the proteolytic cleavage of the viral polyprotein at the junctions of nonstructural proteins downstream of NS3. The NS3 protein was also found to be internally cleaved. In this study, we demonstrated that internal cleavages occurred...
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doi:10.1128/JVI.02712-06
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pub_type: 杂志文章
doi:10.1128/JVI.80.8.3833-3843.2006
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.64.9.4549-4552.1990
更新日期:1990-09-01 00:00:00
abstract::The investigation of papillomavirus chromatin has been hampered by the unavailability of a tissue culture system for vegetative growth of these viruses. We have used, therefore, bovine papillomavirus type 1-transformed hamster embryo fibroblasts containing 200 to 250 episomal genome equivalents per cell as a source of...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.46.2.567-574.1983
更新日期:1983-05-01 00:00:00
abstract::In wild aquatic birds and poultry around the world, influenza A viruses carrying 15 antigenic subtypes of hemagglutinin (HA) and 9 antigenic subtypes of neuraminidase (NA) have been described. Here we describe a previously unidentified antigenic subtype of HA (H16), detected in viruses circulating in black-headed gull...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.79.5.2814-2822.2005
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.69.11.7362-7366.1995
更新日期:1995-11-01 00:00:00
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journal_title:Journal of virology
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doi:10.1128/JVI.69.2.1181-1189.1995
更新日期:1995-02-01 00:00:00
abstract::Human immunodeficiency virus (HIV) infection is associated with immune activation, CD4⁺-T-cell loss, and a progressive decline of immune functions. Antiretroviral therapy (ART) only partially reverses HIV-associated immune dysfunction, suggesting that approaches that target immune activation and improve virus-specific...
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doi:10.1128/JVI.69.1.75-81.1995
更新日期:1995-01-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.61.4.1155-1163.1987
更新日期:1987-04-01 00:00:00
abstract::Human cytomegalovirus (HCMV) gene expression is highly cell and tissue specific. Cell factor-mediated regulatory interactions are involved in regulating the restricted expression of the HCMV major immediate-early (IE) gene (J. F. Baskar, P. P. Smith, G. Nilaver, R. A. Jupp, S. Hoffmann, N. J. Peffer, D. J. Tenney, A. ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.72.1.236-244.1998
更新日期:1998-01-01 00:00:00
abstract::Spleen cells from mice, 4 to 60 days after infection with mouse hepatitis virus type 4, produced interleukin-2, as well as interleukin-3, in the absence of exogenous stimulants in vitro. This unique lymphokine production by mouse hepatitis virus type 4 infection was controlled by host genes. ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.62.9.3506-3508.1988
更新日期:1988-09-01 00:00:00
abstract::To identify the domains of Vpr that are involved nuclear localization, we transfected HeLa cells with a panel of expression vectors that encode mutant Vpr protein with deletions or substitutions within putative domains. Immunofluorescence staining of transfected cells revealed that wild-type Vpr was localized predomin...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.74.15.7179-7186.2000
更新日期:2000-08-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.36.3.860-871.1980
更新日期:1980-12-01 00:00:00
abstract::Analysis of six monoclonal antibody-resistant (mar) mutants in herpes simplex virus type 1 glycoprotein B identified two type-common (II and III) and two type-specific (I and IV) antigenic sites on this molecule. To derive additional information on the location of these sites, mar mutations were mapped and nucleotide ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.63.2.730-738.1989
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.66.2.906-913.1992
更新日期:1992-02-01 00:00:00
abstract::Virus-specific CD4(+) T-cell function is thought to play a central role in induction and maintenance of effective CD8(+) T-cell responses in experimental animals or humans. However, the reasons that diminished proliferation of human immunodeficiency virus (HIV)-specific CD4(+) T cells is observed in the majority of in...
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更新日期:2003-10-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.61.2.594-596.1987
更新日期:1987-02-01 00:00:00
abstract::Papillomavirus disease poses a special challenge to people with compromised immune systems. Appropriate models to study infections in these individuals are lacking. We report here the development of a model that will help to address these deficiencies. The MmuPV1 genome was synthesized and used successfully to produce...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00777-13
更新日期:2013-08-01 00:00:00
abstract::FeLV-FAIDS, an immunodeficiency-inducing isolate of feline leukemia virus, is composed of a pathogenic but replication-defective genome (molecular clone 61C) and a replication-competent but non-immunodeficiency-inducing variant genome (molecular clone 61E). The chimeric virus EECC, composed of the 5' gag-pol of 61E fu...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.64.9.4338-4345.1990
更新日期:1990-09-01 00:00:00
abstract::A protein-protein interaction within the helicase domain of the Tobacco mosaic virus (TMV) 126- and 183-kDa replicase proteins was previously implicated in virus replication (S. Goregaoker, D. Lewandowski, and J. Culver, Virology 282:320-328, 2001). To further characterize the interaction, polypeptides covering the in...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.77.6.3549-3556.2003
更新日期:2003-03-01 00:00:00
abstract::The relatively small package capacity (less than 5 kb) of adeno-associated virus (AAV) vectors has been effectively doubled with the development of dual-vector heterodimerization approaches. However, the efficiency of such dual-vector systems is limited not only by the extent to which intermolecular recombination occu...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.79.1.364-379.2005
更新日期:2005-01-01 00:00:00
abstract::Transcript levels from the P5 promoter of adeno-associated virus type 2 (AAV) are negatively regulated by the AAV Rep78 and Rep68 proteins in the absence of helper virus. We have identified a Rep-responsive negative cis element of the P5 promoter between the P5 TATA box and transcription start site by using 5' and 3' ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.69.11.6787-6796.1995
更新日期:1995-11-01 00:00:00
abstract:UNLABELLED:Venezuelan equine encephalitis virus (VEEV) is an important human and animal pathogen, for which no safe and efficient vaccines or therapeutic means have been developed. Viral particle assembly and budding processes represent potential targets for therapeutic intervention. However, our understanding of the m...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.02680-15
更新日期:2015-12-09 00:00:00
abstract::Hypoxia-inducible factor 1 (HIF-1) is a ubiquitously expressed transcriptional regulator involved in induction of numerous genes associated with angiogenesis and tumor growth. Kaposi's sarcoma, associated with increased angiogenesis, is a highly vascularized, endothelial cell-derived tumor. Previously, we have shown t...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00611-07
更新日期:2007-10-01 00:00:00
abstract::Noroviruses are the causative agents of the majority of viral gastroenteritis outbreaks in humans. During the past 15 years, noroviruses of genotype GGII.4 have caused four epidemic seasons of viral gastroenteritis, during which four novel variants (termed epidemic variants) emerged and displaced the resident viruses....
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00674-07
更新日期:2007-09-01 00:00:00