49 results on '"John W. Schoggins"'
Search Results
2. Enterovirus 3C Protease Cleaves TRIM7 To Dampen Its Antiviral Activity
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Wenchun Fan, Matthew B. McDougal, and John W. Schoggins
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Glutamine ,Ubiquitin-Protein Ligases ,Immunology ,3C Viral Proteases ,Microbiology ,Virus-Cell Interactions ,Tripartite Motif Proteins ,Virology ,Insect Science ,Host-Pathogen Interactions ,Enterovirus Infections ,Animals ,Histidine ,Phascolarctidae ,Enterovirus - Abstract
Mammalian TRIM7 is an antiviral protein that inhibits multiple human enteroviruses by degrading the viral 2BC protein. Whether TRIM7 is reciprocally targeted by enteroviruses is not known. Here, we report that the 3C protease (3Cpro) from two enteroviruses, coxsackievirus B3 (CVB3) and poliovirus, targets TRIM7 for cleavage. CVB3 3Cpro cleaves TRIM7 at glutamine 24 (Q24), resulting in a truncated TRIM7 that fails to inhibit CVB3 due to dampened E3 ubiquitin ligase activity. TRIM7 Q24 is highly conserved across mammals, except in marsupials, which instead have a naturally occurring histidine (H24) that is not subject to 3Cpro cleavage. Marsupials also express two isoforms of TRIM7, and the two proteins from koalas have distinct antiviral activities. The longer isoform contains an additional exon due to alternate splice site usage. This additional exon contains a unique 3Cpro cleavage site, suggesting that certain enteroviruses may have evolved to target marsupial TRIM7 even if the canonical Q24 is missing. Combined with computational analyses indicating that TRIM7 is rapidly evolving, our data raise the possibility that TRIM7 may be targeted by enterovirus evasion strategies and that evolution of TRIM7 across mammals may have conferred unique antiviral properties. IMPORTANCE Enteroviruses are significant human pathogens that cause viral myocarditis, pancreatitis, and meningitis. Knowing how the host controls these viruses and how the viruses may evade host restriction is important for understanding fundamental concepts in antiviral immunity and for informing potential therapeutic interventions. In this study, we demonstrate that coxsackievirus B3 uses its virally encoded protease to target the host antiviral protein TRIM7 for cleavage, suggesting a potential mechanism of viral immune evasion. We additionally show that TRIM7 has evolved in certain mammalian lineages to express protein variants with distinct antiviral activities and susceptibilities to viral protease-mediated cleavage.
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- 2022
3. Identification of interferon-stimulated genes that attenuate Ebola virus infection
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Gabriele Neumann, John W. Schoggins, Charles M. Rice, Peter Halfmann, Makoto Ozawa, Lindsay Hill-Batorski, Makoto Kuroda, Tiago J. S. Lopes, and Yoshihiro Kawaoka
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0301 basic medicine ,viruses ,Fluorescent Antibody Technique ,Gene Expression ,General Physics and Astronomy ,Virus Replication ,medicine.disease_cause ,Ebola virus ,Interferon ,Chlorocebus aethiops ,lcsh:Science ,Multidisciplinary ,Reverse Transcriptase Polymerase Chain Reaction ,virus diseases ,Ebolavirus ,Protein Binding ,medicine.drug ,Cell Survival ,Viral protein ,Science ,Blotting, Western ,030106 microbiology ,Biology ,Virus-host interactions ,Article ,General Biochemistry, Genetics and Molecular Biology ,Virus ,Viral Proteins ,03 medical and health sciences ,Viral entry ,medicine ,Animals ,Humans ,Immunoprecipitation ,Vero Cells ,Gene ,Outbreak ,General Chemistry ,Virus Internalization ,Virology ,Cytoskeletal Proteins ,HEK293 Cells ,030104 developmental biology ,Vero cell ,lcsh:Q ,Interferons ,Restriction factors ,Carrier Proteins ,HeLa Cells - Abstract
The West Africa Ebola outbreak was the largest outbreak ever recorded, with over 28,000 reported infections; this devastating epidemic emphasized the need to understand the mechanisms to counteract virus infection. Here, we screen a library of nearly 400 interferon-stimulated genes (ISGs) against a biologically contained Ebola virus and identify several ISGs not previously known to affect Ebola virus infection. Overexpression of the top ten ISGs attenuates virus titers by up to 1000-fold. Mechanistic studies demonstrate that three ISGs interfere with virus entry, six affect viral transcription/replication, and two inhibit virion formation and budding. A comprehensive study of one ISG (CCDC92) that shows anti-Ebola activity in our screen reveals that CCDC92 can inhibit viral transcription and the formation of complete virions via an interaction with the viral protein NP. Our findings provide insights into Ebola virus infection that could be exploited for the development of therapeutics against this virus., Here, Kuroda et al. screen a library of nearly 400 interferon-stimulated genes (ISGs) and identify several ISGs that inhibit Ebola virus entry, viral transcription/replication, or virion formation. The study provides insights into interactions between Ebola and the host cells.
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- 2020
4. Defective viral RNA sensing linked to severe COVID-19
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John W. Schoggins
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2019-20 coronavirus outbreak ,Multidisciplinary ,Coronavirus disease 2019 (COVID-19) ,SARS-CoV-2 ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,fungi ,food and beverages ,COVID-19 ,Humans ,RNA, Viral ,Viral rna ,Biology ,Virology - Abstract
Genetic variation in a sensor of double-stranded RNA can exacerbate COVID-19
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- 2021
5. Over-expression Screen of Interferon-Stimulated Genes Identifies RARRES3 as a Restrictor ofToxoplasma gondiiInfection
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Michael E. Abrams, Neal M. Alto, Sumit K. Matta, John W. Schoggins, L. David Sibley, and Nicholas Rinkenberger
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medicine.medical_treatment ,Toxoplasma gondii ,Biology ,biology.organism_classification ,Virology ,Cytokine ,IRF1 ,Downregulation and upregulation ,Interferon ,Immunity ,parasitic diseases ,medicine ,Interferon gamma ,Gene ,medicine.drug - Abstract
Toxoplasma gondiiis an important human pathogen infecting an estimated 1 in 3 people worldwide. The cytokine interferon gamma (IFNγ) is induced during infection and is critical for restrictingT. gondiigrowth in human cells. Growth restriction is presumed to be due to the induction interferon stimulated genes (ISGs) that are upregulated to protect the host from infection. Although there are hundreds of ISGs induced by IFNγ, their individual roles in restricting parasite growth in human cells remain somewhat elusive. To address this deficiency, we screened a library of 414 IFNγ induced ISGs to identify factors that impactT. gondiiinfection in human cells. In addition to IRF1, which likely acts through induction of numerous downstream genes, we identified RARRES3 as a single factor that restrictsT. gondiiinfection by inducing premature egress of the parasite in multiple human cell lines. Overall, while we successfully identified a novel IFNγ induced factor restrictingT. gondiiinfection, the limited number of ISGs capable of restrictingT. gondiiinfection when individually expressed suggests that IFNγ mediated immunity toT. gondiiinfection is a complex, multifactorial process.
- Published
- 2021
6. In-Depth Evaluation of a Case of Presumed Myocarditis After the Second Dose of COVID-19 mRNA Vaccine
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Imran Hitto, Mark H. Drazner, Quan Zhen Li, James A. de Lemos, Benjamin Greenberg, Madhusudhanan Narasimhan, John W. Schoggins, Kiran Batra, Amit Khera, Xuan Jiang, Carol L. Croft, Chengsong Zhu, Lenin Mahimainathan, Justin L. Grodin, Franklin Fuda, James B Cutrell, Sean J. Morrison, Pradeep P.A. Mammen, and Alagarraju Muthukumar
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Male ,2019-20 coronavirus outbreak ,Myocarditis ,COVID-19 Vaccines ,Coronavirus disease 2019 (COVID-19) ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,Immunization, Secondary ,Physiology (medical) ,vaccine ,Medicine ,Humans ,Cases and Traces ,Messenger RNA ,business.industry ,SARS-CoV-2 ,COVID-19 ,Middle Aged ,medicine.disease ,Virology ,ComputingMethodologies_DOCUMENTANDTEXTPROCESSING ,Cardiovascular Case Series ,Cardiology and Cardiovascular Medicine ,business ,COVID-19 vaccine ,2019-nCoV Vaccine mRNA-1273 - Abstract
Supplemental Digital Content is available in the text.
- Published
- 2021
7. Comparative analysis of viral entry for Asian and African lineages of Zika virus
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John W. Schoggins and Nicholas Rinkenberger
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Infectivity ,0303 health sciences ,Asia ,Lineage (genetic) ,biology ,Zika Virus Infection ,030302 biochemistry & molecular biology ,Endosomes ,Zika Virus ,Virus Internalization ,biology.organism_classification ,Virology ,Endocytosis ,Article ,Zika virus ,03 medical and health sciences ,Emerging pathogen ,Flavivirus ,Viral entry ,Africa ,Humans ,030304 developmental biology ,Immune activation - Abstract
Zika virus (ZIKV) is an emerging pathogen with global health and economic impacts. ZIKV circulates as two major lineages, Asian or African. The Asian lineage has recently been associated with significant disease in humans. Numerous studies have revealed differences between African and Asian ZIKV strains with respect to cellular infectivity, pathogenesis, and immune activation. Less is known about the mechanism of ZIKV entry and whether viral entry differs between strains. Here, we characterized ZIKV entry with two Asian and two African strains. All viruses exhibited a requirement for clathrin-mediated endocytosis and Rab5a function. Additionally, all ZIKV strains tested were sensitive to pH in the range of 6.5-6.1 and were reliant on endosomal acidification for infection. Finally, we provide direct evidence that ZIKV primarily fuses with late endosomes. These findings contribute new insight into the ZIKV entry process and suggest that divergent ZIKV strains enter cells in a highly conserved manner.
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- 2019
8. Evolution of the interferon response: lessons from ISGs of diverse mammals
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Matthew B McDougal, Ian N Boys, Pamela De La Cruz-Rivera, and John W Schoggins
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Mammals ,Virus Diseases ,Virology ,Viruses ,Animals ,Humans ,Interferons ,Antiviral Agents - Abstract
The vertebrate interferon (IFN) response controls viral infections by inducing hundreds of interferon-stimulated genes (ISGs), many of which encode 'restriction factors' that uniquely target certain viruses. ISG studies have historically had a human-centric focus, which is justified because these natural defense mechanisms might be leveraged to treat human viral disease. However, certain mammals are reservoirs for zoonotic viruses that can 'spill over' into humans. Additionally, restriction factors have prominent roles in the ongoing evolutionary genetic conflicts between viruses and their hosts. Thus, there is a growing need to understand antiviral IFN/ISG responses in other species, particularly in known reservoirs of zoonotic viruses. This review focuses on functional and evolutionary insight into antiviral IFN responses that have been obtained from studying non-model mammalian species.
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- 2022
9. Laboratory Action Plan for Emerging SARS-CoV-2 Variants
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Jason Y. Park, Jeffrey A. SoRelle, Laura M. Filkins, and John W. Schoggins
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Biochemistry, medical ,2019-20 coronavirus outbreak ,Opinion ,Coronavirus disease 2019 (COVID-19) ,SARS-CoV-2 ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,Biochemistry (medical) ,Clinical Biochemistry ,COVID-19 ,Biology ,Virology ,COVID-19 Serological Testing ,Coronavirus Disease 2019 ,Action plan ,Humans ,VUI-202012/01 ,Laboratories ,B.1.1.7 ,VOC-202012/01 - Published
- 2021
10. TRIM7 inhibits enterovirus replication and promotes emergence of a viral variant with increased pathogenicity
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John M. Shelton, Wenchun Fan, Bret M. Evers, Milo M. Lin, Ilona K. Gaszek, Katrina B. Mar, Levent Sari, Qiang Cheng, Mary Wight-Carter, Daniel J. Siegwart, and John W. Schoggins
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Proteasome Endopeptidase Complex ,Viral pathogenesis ,viruses ,Ubiquitin-Protein Ligases ,Coxsackievirus ,Virus Replication ,General Biochemistry, Genetics and Molecular Biology ,Virus ,Article ,Cell Line ,Tripartite Motif Proteins ,03 medical and health sciences ,Viral Proteins ,0302 clinical medicine ,Interferon ,medicine ,Animals ,Humans ,030304 developmental biology ,Enterovirus ,Adenosine Triphosphatases ,Inflammation ,0303 health sciences ,biology ,Effector ,Ubiquitin ,biology.organism_classification ,Virology ,Ubiquitin ligase ,Mice, Inbred C57BL ,Viral replication ,Viral evolution ,Mutation ,Proteolysis ,biology.protein ,RNA, Viral ,Female ,030217 neurology & neurosurgery ,medicine.drug ,Protein Binding - Abstract
To control viral infection, vertebrates rely on both inducible interferon responses and less well characterized cell intrinsic responses comprised of “at the ready” antiviral effector proteins. Here, we show that E3 ubiquitin ligase TRIM7 is a cell intrinsic antiviral effector that restricts multiple human enteroviruses by targeting viral 2BC, a membrane remodeling protein, for ubiquitination and proteasome-dependent degradation. Selective pressure exerted by TRIM7 results in emergence of a TRIM7-resistant coxsackievirus with a single point mutation in the viral 2C ATPase/helicase. In cultured cells, the mutation helps the virus evade TRIM7, but impairs optimal viral replication, and this correlates with a hyperactive and structurally plastic 2C ATPase. Unexpectedly, the TRIM7-resistant virus has a replication advantage in mice and causes lethal pancreatitis. These findings reveal a unique mechanism for targeting enterovirus replication and provide molecular insight into the benefits and trade-offs of viral evolution imposed by a host restriction factor.
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- 2020
11. A Sensitive Yellow Fever Virus Entry Reporter Identifies Valosin-Containing Protein (VCP/p97) as an Essential Host Factor for Flavivirus Uncoating
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Brett D. Lindenbach, John W. Schoggins, Shuo Zhang, Harish N. Ramanathan, Jinhong Chang, Priscilla L. Yang, Katrina B. Mar, Alexander Ploss, and Florian Douam
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Valosin-containing protein ,viruses ,nucleocapsid ,lcsh:A ,Genome, Viral ,Dengue virus ,Kidney ,Virus Replication ,medicine.disease_cause ,Microbiology ,Virus ,Cell Line ,Host-Microbe Biology ,Mice ,flavivirus ,Genes, Reporter ,Valosin Containing Protein ,Viral entry ,Cricetinae ,Virus Uncoating ,Virology ,ubiquitin ,medicine ,Animals ,Humans ,biology ,VCP/p97 ,RNA ,Virus Internalization ,biology.organism_classification ,QR1-502 ,Mice, Inbred C57BL ,Flavivirus ,HEK293 Cells ,Capsid ,biology.protein ,Female ,viral entry ,uncoating ,lcsh:General Works ,Yellow fever virus ,RNA transfection ,HeLa Cells ,Research Article - Abstract
Flaviviruses are an important group of RNA viruses that cause significant human disease. The mechanisms by which flavivirus nucleocapsids are disassembled during virus entry remain unclear. Here, we used a yellow fever virus entry reporter, which expresses a sensitive reporter enzyme but does not replicate, to show that nucleocapsid disassembly requires the cellular protein-disaggregating enzyme valosin-containing protein, also known as p97., While the basic mechanisms of flavivirus entry and fusion are understood, little is known about the postfusion events that precede RNA replication, such as nucleocapsid disassembly. We describe here a sensitive, conditionally replication-defective yellow fever virus (YFV) entry reporter, YFVΔSK/Nluc, to quantitively monitor the translation of incoming, virus particle-delivered genomes. We validated that YFVΔSK/Nluc gene expression can be neutralized by YFV-specific antisera and requires known flavivirus entry pathways and cellular factors, including clathrin- and dynamin-mediated endocytosis, endosomal acidification, YFV E glycoprotein-mediated fusion, and cellular LY6E and RPLP1 expression. The initial round of YFV translation was shown to require cellular ubiquitylation, consistent with recent findings that dengue virus capsid protein must be ubiquitylated in order for nucleocapsid uncoating to occur. Importantly, translation of incoming YFV genomes also required valosin-containing protein (VCP)/p97, a cellular ATPase that unfolds and extracts ubiquitylated client proteins from large complexes. RNA transfection and washout experiments showed that VCP/p97 functions at a postfusion, pretranslation step in YFV entry. Finally, VCP/p97 activity was required by other flaviviruses in mammalian cells and by YFV in mosquito cells. Together, these data support a critical role for VCP/p97 in the disassembly of incoming flavivirus nucleocapsids during a postfusion step in virus entry.
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- 2020
12. LY6E impairs coronavirus fusion and confers immune control of viral disease
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Gert Zimmer, Stephanie Pfaender, Daniel Todt, Wenchun Fan, Tom Gallagher, Eleftherios Michailidis, Hans Heinrich Hoffmann, Katrina B. Mar, Annika Kratzel, Philip V'kovski, Nadine Ebert, Matthew B. McDougal, Ronald Dijkman, Natasha W. Hanners, Mohsan Saeed, Volker Thiel, Jenna N. Kelly, Mary Wight-Carter, Charles M. Rice, Hannah Kleine-Weber, Dagny Hirt, Ian N. Boys, Hanspeter Stalder, Eike Steinmann, Markus Hoffmann, John W. Schoggins, and Stefan Pöhlmann
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Male ,viruses ,Viral pathogenesis ,medicine.disease_cause ,Applied Microbiology and Biotechnology ,Pathogenesis ,Mice ,0302 clinical medicine ,Coronavirus ,Mice, Knockout ,0303 health sciences ,virus diseases ,respiratory system ,3. Good health ,Haematopoiesis ,Severe acute respiratory syndrome-related coronavirus ,030220 oncology & carcinogenesis ,Antigens, Surface ,Knockout mouse ,Middle East Respiratory Syndrome Coronavirus ,Female ,Angiotensin-Converting Enzyme 2 ,Viral disease ,Coronavirus Infections ,Viral load ,Microbiology (medical) ,Middle East respiratory syndrome coronavirus ,Pneumonia, Viral ,Immunology ,Peptidyl-Dipeptidase A ,Biology ,GPI-Linked Proteins ,Microbiology ,Article ,Betacoronavirus ,03 medical and health sciences ,Immune system ,Antigen ,Genetics ,medicine ,Animals ,Pandemics ,030304 developmental biology ,Innate immune system ,SARS-CoV-2 ,COVID-19 ,Cell Biology ,Virus Internalization ,biochemical phenomena, metabolism, and nutrition ,medicine.disease ,Virology ,respiratory tract diseases ,Mice, Inbred C57BL ,Middle East respiratory syndrome - Abstract
Zoonotic coronaviruses (CoVs) are significant threats to global health, as exemplified by the recent emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)1. Host immune responses to CoV are complex and regulated in part through antiviral interferons. However, the interferon-stimulated gene products that inhibit CoV are not well characterized2. Here, we show that interferon-inducible lymphocyte antigen 6 complex, locus E (LY6E) potently restricts cellular infection by multiple CoVs, including SARS-CoV, SARS-CoV-2, and Middle East respiratory syndrome coronavirus (MERS-CoV). Mechanistic studies revealed that LY6E inhibits CoV entry into cells by interfering with spike protein-mediated membrane fusion. Importantly, mice lacking Ly6e in hematopoietic cells were highly susceptible to murine CoV infection. Exacerbated viral pathogenesis in Ly6e knockout mice was accompanied by loss of hepatic and splenic immune cells and reduction in global antiviral gene pathways. Accordingly, we found that Ly6e directly protects primary B cells and dendritic cells from murine CoV infection. Our results demonstrate that LY6E is a critical antiviral immune effector that controls CoV infection and pathogenesis. These findings advance our understanding of immune-mediated control of CoV in vitro and in vivo, knowledge that could help inform strategies to combat infection by emerging CoV.
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- 2020
13. The immune vulnerability landscape of the 2019 Novel Coronavirus, SARS-CoV-2
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Yunguan Wang, Lenny Moise, He Zhang, Yang Xie, Xue Xiao, Xiaowei Zhan, James Zhu, John W. Schoggins, Jiwoong Kim, Shuang Jiang, Lin Xu, Tao Wang, Ran Chen, Danni Luo, Guanghua Xiao, Andres H. Gutierrz, and Anne S. De Groot
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B cell ,SARS-CoV-2 ,viruses ,T cell ,Immunogenicity ,virus diseases ,immunogenicity ,biochemical phenomena, metabolism, and nutrition ,Biology ,Virology ,Article ,Virus ,Epitope ,respiratory tract diseases ,medicine.anatomical_structure ,Immune system ,Immunity ,medicine ,mutation ,skin and connective tissue diseases ,CD8 - Abstract
The outbreak of the 2019 Novel Coronavirus (SARS-CoV-2) rapidly spread from Wuhan, China to more than 150 countries, areas, or territories, causing staggering numbers of infections and deaths. In this study, bioinformatics analyses were performed on 5,568 complete genomes of SARS-CoV-2 virus to predict the T cell and B cell immunogenic epitopes of all viral proteins, which formed a systematic immune vulnerability landscape of SARS-CoV-2. The immune vulnerability and genetic variation profiles of SARS-CoV were compared with those of SARS-CoV and MERS-CoV. In addition, a web portal was developed to broadly share the data and results as a resource for the research community. Using this resource, we showed that genetic variations in SARS-CoV-2 are associated with loss of B cell immunogenicity, an increase in CD4+ T cell immunogenicity, and a minimum loss in CD8+ T cell immunogenicity, indicating the existence of a curious correlation between SARS-CoV-2 genetic evolutions and the immunity pressure from the host. Overall, we present an immunological resource for SARS-CoV-2 that could promote both therapeutic/vaccine development and mechanistic research.
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- 2020
14. The IFN Response in Bats Displays Distinctive IFN-Stimulated Gene Expression Kinetics with Atypical RNASEL Induction
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Chao Xing, John W. Schoggins, Mohammed Kanchwala, Lin-Fa Wang, Hanquan Liang, Pamela C. De La Cruz-Rivera, and Ashwani Kumar
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0301 basic medicine ,Regulation of gene expression ,Disease reservoir ,Innate immune system ,biology ,Effector ,Immunology ,virus diseases ,biology.organism_classification ,Virology ,03 medical and health sciences ,030104 developmental biology ,Cell culture ,Immunity ,Immunology and Allergy ,Pteropus alecto ,Gene - Abstract
Bats host a large number of zoonotic viruses, including several viruses that are highly pathogenic to other mammals. The mechanisms underlying this rich viral diversity are unknown, but they may be linked to unique immunological features that allow bats to act as asymptomatic viral reservoirs. Vertebrates respond to viral infection by inducing IFNs, which trigger antiviral defenses through IFN-stimulated gene (ISG) expression. Although the IFN system of several bats is characterized at the genomic level, less is known about bat IFN-mediated transcriptional responses. In this article, we show that IFN signaling in bat cells from the black flying fox (Pteropus alecto) consists of conserved and unique ISG expression profiles. In IFN-stimulated cells, bat ISGs comprise two unique temporal subclusters with similar early induction kinetics but distinct late-phase declines. In contrast, human ISGs lack this decline phase and remained elevated for longer periods. Notably, in unstimulated cells, bat ISGs were expressed more highly than their human counterparts. We also found that the antiviral effector 2-5A–dependent endoribonuclease, which is not an ISG in humans, is highly IFN inducible in black flying fox cells and contributes to cell-intrinsic control of viral infection. These studies reveal distinctive innate immune features that may underlie a unique virus–host relationship in bats.
- Published
- 2018
15. Genetic Variation at IFNL4 Influences Extrahepatic Interferon-Stimulated Gene Expression in Chronic HCV Patients
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Brad R. Rosenberg, Spencer T. Chen, Rachel Caron, Charles M. Rice, John W. Schoggins, Naoko Imanaka, Andrew H. Talal, Jennifer L. Eitson, Skyler Uhl, Marija Zeremski, Ira M. Jacobson, and Catherine A. Freije
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0301 basic medicine ,Hepatitis C virus ,Alpha interferon ,Biology ,medicine.disease_cause ,Peripheral blood mononuclear cell ,Major Articles and Brief Reports ,03 medical and health sciences ,Interferon ,Gene expression ,medicine ,Humans ,Immunologic Factors ,Immunology and Allergy ,Regulation of gene expression ,Blood Cells ,Sequence Analysis, RNA ,Gene Expression Profiling ,Interleukins ,Interferon-stimulated gene ,Genetic Variation ,Interferon-alpha ,virus diseases ,Hepatitis C ,Hepatitis C, Chronic ,medicine.disease ,Virology ,030104 developmental biology ,Infectious Diseases ,Gene Expression Regulation ,Immunology ,Leukocytes, Mononuclear ,medicine.drug - Abstract
Polymorphisms at IFNL4 strongly influence spontaneous resolution and interferon therapeutic response in hepatitis C virus (HCV) infection. In chronic HCV, unfavorable alleles are associated with elevated interferon (IFN)-stimulated gene (ISG) expression in the liver, but extrahepatic effects are less well characterized. We used RNA sequencing (RNA-Seq) to examine whether IFNL4 genetic variation (rs368234815) modulates ISG expression in peripheral blood mononuclear cells (PBMC) during chronic HCV infection. ISG expression was elevated in unstimulated PBMC homozygous for the unfavorable ΔG IFNL4 variant; expression following IFN-α stimulation was comparable across genotypes. These findings suggest that lambda interferons may have broader systemic effects during HCV infection.
- Published
- 2017
16. The cellular endosomal protein stannin inhibits intracellular trafficking of human papillomavirus during virus entry
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Asu Erden, John W. Schoggins, Alex Lipovsky, Akiko Iwasaki, John D. MacMicking, Clinton J. Bradfield, Daniel DiMaio, Wei Zhang, Mac Crite, and Eriko Kanaya
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Keratinocytes ,0301 basic medicine ,Retromer ,Endosome ,viruses ,Biology ,Virus ,Cell Line ,03 medical and health sciences ,Viral entry ,Virology ,Animals ,Humans ,Gene ,Human papillomavirus 16 ,Neuropeptides ,Virus Uncoating ,virus diseases ,Oncogene Proteins, Viral ,Virus Internalization ,female genital diseases and pregnancy complications ,Cell biology ,030104 developmental biology ,Capsid ,Capsid Proteins ,Intracellular ,Protein Binding ,Research Article - Abstract
Human papillomaviruses (HPVs) are the most common sexually transmitted viruses and one of the most important infectious causes of cancers worldwide. While prophylactic vaccines are effective against certain strains of HPV, established infections still cause deadly cancers in both men and women. HPV traffics to the nucleus via the retrograde transport pathway, but the mechanism of intracellular transport of non-enveloped viruses such as HPV is incompletely understood. Using an overexpression screen, we identify several genes that control HPV16 entry. We focused on the mechanism by which one of the screen hits, stannin, blocks HPV16 infection. Stannin has not been previously implicated in virus entry. Overexpression of stannin specifically inhibits infection by several HPV types, but not other viruses tested. Stannin is constitutively expressed in human keratinocytes, and its basal levels limit entry by HPV16. Stannin is localized to the endolysosomal compartment and does not affect HPV16 binding to cells, virus uptake, or virus uncoating, but inhibits the entry of HPV into the trans-Golgi network (TGN) and stimulates HPV degradation. We further show that stannin interacts with L1 major capsid protein and impairs the interaction of the L2 minor capsid protein with retromer, which is required for virus trafficking to the TGN. Our findings shed light on a novel cellular protein that interferes with HPV entry and highlight the role of retrograde transport in HPV entry.
- Published
- 2017
17. Functional-genomic analysis reveals intraspecies diversification of antiviral receptor transporter proteins in Xenopus laevis
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Ian N. Boys, Katrina B. Mar, and John W. Schoggins
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RNA viruses ,0106 biological sciences ,Cancer Research ,African clawed frog ,Xenopus ,Xenopus Proteins ,QH426-470 ,01 natural sciences ,Xenopus laevis ,Spectrum Analysis Techniques ,Medical Conditions ,Medicine and Health Sciences ,Genetics (clinical) ,Data Management ,Mammals ,Genetics ,0303 health sciences ,biology ,Effector ,Eukaryota ,Vertebrate ,Phylogenetic Analysis ,Genomics ,Animal Models ,Flow Cytometry ,Phylogenetics ,Infectious Diseases ,Experimental Organism Systems ,Spectrophotometry ,Viral evolution ,Vertebrates ,Viruses ,Frogs ,Cytophotometry ,Research Article ,Computer and Information Sciences ,Infectious Disease Control ,Protein family ,Research and Analysis Methods ,Antiviral Agents ,Synteny ,Microbiology ,010603 evolutionary biology ,Evolution, Molecular ,Amphibians ,03 medical and health sciences ,Model Organisms ,Virology ,biology.animal ,Animals ,Evolutionary Systematics ,Molecular Biology ,Ecology, Evolution, Behavior and Systematics ,Taxonomy ,030304 developmental biology ,Evolutionary Biology ,Organisms ,Membrane Transport Proteins ,Biology and Life Sciences ,RNA ,biology.organism_classification ,Viral Replication ,Poly I-C ,Amniotes ,Animal Studies ,Zoology ,Function (biology) - Abstract
The Receptor Transporter Protein (RTP) family is present in most, if not all jawed vertebrates. Most of our knowledge of this protein family comes from studies on mammalian RTPs, which are multi-function proteins that regulate cell-surface G-protein coupled receptor levels, influence olfactory system development, regulate immune signaling, and directly inhibit viral infection. However, mammals comprise less than one-tenth of extant vertebrate species, and our knowledge about the expression, function, and evolution of non-mammalian RTPs is limited. Here, we explore the evolutionary history of RTPs in vertebrates. We identify signatures of positive selection in many vertebrate RTP clades and characterize multiple, independent expansions of the RTP family outside of what has been described in mammals. We find a striking expansion of RTPs in the African clawed frog, Xenopus laevis, with 11 RTPs in this species as opposed to 1 to 4 in most other species. RNA sequencing revealed that most X. laevis RTPs are upregulated following immune stimulation. In functional assays, we demonstrate that at least three of these X. laevis RTPs inhibit infection by RNA viruses, suggesting that RTP homologs may serve as antiviral effectors outside of Mammalia., Author summary Receptor Transporter Proteins (RTPs) are a family of proteins which, in mammals, are involved in diverse physiological processes such as olfaction and innate immunity. Less, however, is known about non-mammalian RTPs. In this study, we investigate the evolutionary history of RTPs and identify antiviral RTPs in the African clawed frog, Xenopus laevis.
- Published
- 2021
18. Interferon-Stimulated Genes: What Do They All Do?
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John W. Schoggins
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Biology ,Virus ,03 medical and health sciences ,0302 clinical medicine ,Interferon ,Virology ,medicine ,Animals ,Humans ,Gene ,030304 developmental biology ,0303 health sciences ,Innate immune system ,virus diseases ,Acquired immune system ,Cell biology ,Virus Diseases ,Interferon Regulatory Factors ,Viruses ,Human genome ,Interferons ,Signal transduction ,030217 neurology & neurosurgery ,Function (biology) ,medicine.drug ,Virus Physiological Phenomena - Abstract
In the absence of an intact interferon (IFN) response, mammals may be susceptible to lethal viral infection. IFNs are secreted cytokines that activate a signal transduction cascade leading to the induction of hundreds of interferon-stimulated genes (ISGs). Remarkably, approximately 10% of the genes in the human genome have the potential to be regulated by IFNs. What do all of these genes do? It is a complex question without a simple answer. From decades of research, we know that many of the protein products encoded by these ISGs work alone or in concert to achieve one or more cellular outcomes, including antiviral defense, antiproliferative activities, and stimulation of adaptive immunity. The focus of this review is the antiviral activities of the IFN/ISG system. This includes general paradigms of ISG function, supported by specific examples in the literature, as well as methodologies to identify and characterize ISG function.
- Published
- 2019
19. RTP4 Is a Potent IFN-Inducible Anti-flavivirus Effector Engaged in a Host-Virus Arms Race in Bats and Other Mammals
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John W. Schoggins, Jennifer L. Eitson, Ian N. Boys, Pamela C. De La Cruz-Rivera, Katrina B. Mar, Benjamin Moon, and Elaine Xu
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Male ,viruses ,RNA-dependent RNA polymerase ,Human pathogen ,Genome, Viral ,Virus Replication ,Antiviral Agents ,Microbiology ,Virus ,Cell Line ,03 medical and health sciences ,Flaviviridae ,0302 clinical medicine ,Species Specificity ,Interferon ,Chiroptera ,Virology ,medicine ,Animals ,Humans ,030304 developmental biology ,Mammals ,0303 health sciences ,biology ,Host (biology) ,Effector ,Flavivirus ,virus diseases ,biochemical phenomena, metabolism, and nutrition ,Preview ,biology.organism_classification ,Host-Pathogen Interactions ,Viruses ,Female ,Parasitology ,Interferons ,030217 neurology & neurosurgery ,Molecular Chaperones ,medicine.drug - Abstract
Summary Among mammals, bats are particularly rich in zoonotic viruses, including flaviviruses. Certain bat species can be productively yet asymptomatically infected with viruses that cause overt disease in other species. However, little is known about the antiviral effector repertoire in bats relative to other mammals. Here, we report the black flying fox receptor transporter protein 4 (RTP4) as a potent interferon (IFN)-inducible inhibitor of human pathogens in the Flaviviridae family, including Zika, West Nile, and hepatitis C viruses. Mechanistically, RTP4 associates with the flavivirus replicase, binds viral RNA, and suppresses viral genome amplification. Comparative approaches revealed that RTP4 undergoes positive selection, that a flavivirus can mutate to escape RTP4-imposed restriction, and that diverse mammalian RTP4 orthologs exhibit striking patterns of specificity against distinct Flaviviridae members. Our findings reveal an antiviral mechanism that has likely adapted over 100 million years of mammalian evolution to accommodate unique host-virus genetic conflicts.
- Published
- 2020
20. Western Zika Virus in Human Fetal Neural Progenitors Persists Long Term with Partial Cytopathic and Limited Immunogenic Effects
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Eric M. Wexler, R. Blake Richardson, Genevieve Konopka, John W. Schoggins, Jennifer L. Eitson, Natasha W. Hanners, and Noriyoshi Usui
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0301 basic medicine ,Intrinsic immunity ,Microcephaly ,Cell type ,Time Factors ,Biology ,Virus Replication ,Article ,General Biochemistry, Genetics and Molecular Biology ,Cell Line ,Zika virus ,03 medical and health sciences ,Fetus ,0302 clinical medicine ,Cytopathogenic Effect, Viral ,Neural Stem Cells ,medicine ,Humans ,Progenitor cell ,lcsh:QH301-705.5 ,Zika Virus Infection ,Zika Virus ,medicine.disease ,biology.organism_classification ,Virology ,3. Good health ,030104 developmental biology ,Viral replication ,lcsh:Biology (General) ,Immunology ,030217 neurology & neurosurgery ,Pyknosis - Abstract
SUMMARY The recent Zika virus (ZIKV) outbreak in the Western hemisphere is associated with severe pathology in newborns, including microcephaly and brain damage. The mechanisms underlying these outcomes are under intense investigation. Here, we show that a 2015 ZIKV isolate replicates in multiple cell types, including primary human fetal neural progenitors (hNPs). In immortalized cells, ZIKV is cytopathic and grossly rearranges endoplasmic reticulum membranes similar to other flaviviruses. In hNPs, ZIKV infection has a partial cytopathic phase characterized by cell rounding, pyknosis, and activation of caspase 3. Despite notable cell death, ZIKV did not activate a cytokine response in hNPs. This lack of cell intrinsic immunity to ZIKV is consistent with our observation that virus replication persists in hNPs for at least 28 days. These findings, supported by published fetal neuropathology, establish a proof-of-concept that neural progenitors in the developing human fetus can be direct targets of detrimental ZIKV-induced pathology., In Brief Hanners et al. establish a cell-culture model of a 2015 ZIKV isolate in primary human fetal neural progenitors. They show that ZIKV infection kills a subset of fetal neural progenitors with limited activation of inflammatory or immune responses. In the surviving neural progenitors, ZIKV replication persists for many weeks, mirroring fetal neuropathological findings in vivo.
- Published
- 2016
21. Identification of Interferon-Stimulated Genes with Antiretroviral Activity
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John W. Schoggins, Trinity Zang, Fengwen Zhang, Tonya Kueck, Melissa Kane, Mudathir Alim, Suzannah J. Rihn, Charles M. Rice, Paul D. Bieniasz, and Sam J. Wilson
- Subjects
0301 basic medicine ,Resource ,Cancer Research ,Ubiquitin-Protein Ligases ,Virus Replication ,Microbiology ,Macaque ,Antiviral Agents ,03 medical and health sciences ,Interferon ,Immunology and Microbiology(all) ,Virology ,biology.animal ,medicine ,Animals ,Humans ,Indoleamine-Pyrrole 2,3,-Dioxygenase ,Genomic library ,Genetic Testing ,Molecular Biology ,Gene ,Host protein ,Gene Library ,030102 biochemistry & molecular biology ,biology ,virus diseases ,Macaca mulatta ,3. Good health ,030104 developmental biology ,Retroviridae ,Viral replication ,Parasitology ,Interferons ,medicine.drug - Abstract
Summary Interferons (IFNs) exert their anti-viral effects by inducing the expression of hundreds of IFN-stimulated genes (ISGs). The activity of known ISGs is insufficient to account for the antiretroviral effects of IFN, suggesting that ISGs with antiretroviral activity are yet to be described. We constructed an arrayed library of ISGs from rhesus macaques and tested the ability of hundreds of individual macaque and human ISGs to inhibit early and late replication steps for 11 members of the retroviridae from various host species. These screens uncovered numerous ISGs with antiretroviral activity at both the early and late stages of virus replication. Detailed analyses of two antiretroviral ISGs indicate that indoleamine 2,3-dioxygenase 1 (IDO1) can inhibit retroviral replication by metabolite depletion while tripartite motif-56 (TRIM56) accentuates ISG induction by IFNα and inhibits the expression of late HIV-1 genes. Overall, these studies reveal numerous host proteins that mediate the antiretroviral activity of IFNs., Graphical Abstract Image 1, Highlights • ISG screening identifies direct and indirect antiretroviral proteins • Interferon-γ inhibits HIV-1 through IDO1-mediated tryptophan depletion • TRIM56 enhances the antiretroviral potential of interferon-α, Screening of interferon-stimulated genes for antiretroviral activity reveals numerous genes that directly or indirectly inhibit retroviral replication. Detailed analyses of two antiretroviral effectors indicate that IDO1 inhibits retroviral replication via nutrient depletion while TRIM56 increases the antiretroviral potential of IFNα.
- Published
- 2016
22. Mucolipin-2 Cation Channel Increases Trafficking Efficiency of Endocytosed Viruses
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Nicholas Rinkenberger, John W. Schoggins, and Nancy C. Reich
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0301 basic medicine ,Endosome ,viruses ,virus entry ,Endocytosis ,medicine.disease_cause ,Microbiology ,Virus ,Cell Line ,03 medical and health sciences ,Transient Receptor Potential Channels ,flavivirus ,Viral life cycle ,Viral entry ,Virology ,Influenza A virus ,medicine ,endocytosis ,Humans ,virus-host interactions ,Host factor ,biology ,ion channels ,Zika Virus ,Virus Internalization ,biology.organism_classification ,QR1-502 ,3. Good health ,Cell biology ,Flavivirus ,030104 developmental biology ,Host-Pathogen Interactions ,Yellow fever virus ,influenza ,vesicular trafficking ,Research Article - Abstract
Receptor-mediated endocytosis is a cellular process commonly hijacked by viruses to enter cells. The stages of entry are well described for certain viruses, but the host factors that mediate each step are less well characterized. We previously identified endosomal cation channel mucolipin-2 (MCOLN2) as a host factor that promotes viral infection. Here, we assign a role for MCOLN2 in modulating viral entry. We show that MCOLN2 specifically promotes viral vesicular trafficking and subsequent escape from endosomal compartments. This mechanism requires channel activity, occurs independently of antiviral signaling, and broadly applies to enveloped RNA viruses that require transport to late endosomes for infection, including influenza A virus, yellow fever virus, and Zika virus. We further identify a rare allelic variant of human MCOLN2 that has a loss-of-function phenotype with respect to viral enhancement. These findings establish a mechanistic link between an endosomal cation channel and late stages of viral entry., IMPORTANCE Viruses must co-opt cellular processes to complete their life cycle. To enter cells, viruses frequently take advantage of cellular receptor-mediated endocytosis pathways. A growing number of host proteins are implicated in these viral uptake pathways. Here, we describe a new role for the gated cation channel MCOLN2 in viral entry. This endosomal protein modulates viral entry by enhancing the efficiency of viral trafficking through the endosomal system. Thus, MCOLN2-mediated enhancement of infection may represent a key vulnerability in the viral life cycle that could be targeted for therapeutic intervention.
- Published
- 2018
23. The Cellular Endosomal Protein Stannin Inhibits Human Papillomavirus Entry
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Mac Crite, Alex Lipovsky, Clinton J. Bradfield, Daniel DiMaio, Wei Zhang, John D. MacMicking, Asu Erden, Akiko Iwasaki, John W. Schoggins, and Eriko Kanaya
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Retromer ,Endosome ,viruses ,virus diseases ,Biology ,Virology ,female genital diseases and pregnancy complications ,medicine.anatomical_structure ,Viral envelope ,Capsid ,Viral entry ,Lysosome ,Axoplasmic transport ,medicine ,Compartment (development) - Abstract
Human papillomaviruses (HPVs) are the most common sexually transmitted viruses and one of the most important infectious causes of cancers worldwide. While prophylactic vaccines are effective against certain strains of HPV, established infections still cause deadly cancers in both men and women. HPV traffics to the nucleus via the retrograde transport pathway, but the mechanism of intracellular transport of nonenveloped viruses such as HPV is incompletely understood. Using an overexpression screen, we identify several novel genes that control HPV16 entry. We focused on the mechanism by which one of the screen hits, stannin, blocks HPV16 infection. Stannin has not been previously implicated in virus entry. We show that stannin is constitutively expressed in human keratinocytes, and its basal levels inhibit entry by HPV16. Overexpression of stannin specifically inhibits infection by several HPV types, but not other non-enveloped viruses tested. Stannin is localized to the endolysosomal compartment and does not affect HPV16 binding to cells, uptake, or uncoating, but diverts HPV away from the trans-Golgi network and directs it to the lysosome where it is degraded. We further show that SNN interacts with L1 major capsid protein and that this interaction is modulated by the retromer binding motifs in the C-terminus of the minor capsid protein L2. Our findings shed light on novel cellular factors that control HPV entry.
- Published
- 2018
24. The IFN response in bat cells consists of canonical and non-canonical ISGs with unique temporal expression kinetics
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Ashwani Kumar, Pamela C. De La Cruz-Rivera, Mohammed Kanchwala, John W. Schoggins, Hanquan Liang, Chao Xing, and Lin-Fa Wang
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Genetics ,0303 health sciences ,Innate immune system ,biology ,Ifn response ,030302 biochemistry & molecular biology ,Vertebrate ,virus diseases ,biology.organism_classification ,Virology ,03 medical and health sciences ,Non canonical ,Interferon ,biology.animal ,medicine ,Transcriptional response ,Pteropus alecto ,Gene ,030304 developmental biology ,medicine.drug - Abstract
Bats are reservoirs for a number of highly pathogenic zoonotic viruses, yet they remain relatively asymptomatic during infection. Whether this viral resistance is due to a unique innate immune system is unknown. An evolutionarily conserved feature of vertebrate antiviral immunity is the interferon (IFN) response, which triggers cellular defenses through interferon-stimulated gene (ISG) expression. While bats encode an intact IFN system, global ISG expression patterns in bat cells are not well characterized. Here, we used RNA-Seq to assess the transcriptional response to IFNα in cells derived from the batPteropus alecto(black flying fox). We show induction of more than 100 transcripts, most of which are canonical ISGs observed in other species. Kinetic gene profiling revealed thatP. alectoISGs fall into two unique temporal subclusters with similar early induction kinetics but distinct late-phase declines. In contrast to bat ISGs, human ISGs generally remained elevated for longer periods following IFN treatment, suggesting host-based differences in gene regulatory mechanisms. Notably, we also identified a small group of non-canonical bat ISGs, including an enzymatically active RNASEL that plays a role in controlling viral infection. These studies provide insight into the innate immune response of an important viral reservoir and lay a foundation for studies into the immunological features that may underlie unique virus-host relationship in bats.Significance StatementBats are considered unique in their ability to resist disease caused by viruses that are often pathogenic in humans. While the nature of this viral resistance is unknown, genomic data suggest bat innate immune systems may be specialized in controlling these disease-causing viruses. A critical cell intrinsic antiviral defense system in vertebrates is the interferon response, which suppresses viral infection through induction of hundreds of interferon-stimulated genes (ISGs). In this study, we report the repertoire of ISGs and several unique features of ISG induction kinetics in bat cells. We also characterize induction and antiviral activity of bat RNASEL, which is induced by IFN in bat, but not human cells. These studies lay the foundation for discovery of potentially new antiviral mechanisms in bats, which may spur research into development of therapies to combat viral infection.
- Published
- 2017
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25. Interferon-stimulated genes: roles in viral pathogenesis
- Author
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John W. Schoggins
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Regulation of gene expression ,Extramural ,viruses ,Viral pathogenesis ,virus diseases ,Biology ,Virology ,Article ,Virus ,Gene Expression Regulation ,Virus Diseases ,Interferon ,Host-Pathogen Interactions ,Viruses ,Immunology ,Knockout mouse ,medicine ,Animals ,Humans ,Interferons ,Viral disease ,Gene ,medicine.drug - Abstract
Interferon-stimulated genes (ISGs) are critical for controlling virus infections. As new antiviral ISGs continue to be identified and characterized, their roles in viral pathogenesis are also being explored in more detail. Our current understanding of how ISGs impact viral pathogenesis comes largely from studies in knockout mice, with isolated examples from human clinical data. This review outlines recent developments on the contributions of various ISGs to viral disease outcomes in vivo.
- Published
- 2014
26. Inhibition of HIV-1 Particle Assembly by 2′,3′-Cyclic-Nucleotide 3′-Phosphodiesterase
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Charles M. Rice, Mudathir Alim, John W. Schoggins, Sebla B. Kutluay, Nolwenn Jouvenet, Paul D. Bieniasz, Julia Bitzegeio, Trinity Zang, and Sam J. Wilson
- Subjects
Cancer Research ,Sequence analysis ,viruses ,Molecular Sequence Data ,Sequence alignment ,Plasma protein binding ,Microbiology ,gag Gene Products, Human Immunodeficiency Virus ,Article ,2',3'-Cyclic-nucleotide 3'-phosphodiesterase ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,0302 clinical medicine ,2',3'-Cyclic Nucleotide 3'-Phosphodiesterase ,Virology ,Immunology and Microbiology(all) ,Animals ,Humans ,Amino Acid Sequence ,Peptide sequence ,Gene ,Molecular Biology ,Cells, Cultured ,030304 developmental biology ,0303 health sciences ,biology ,Virus Assembly ,virus diseases ,Sequence Analysis, DNA ,biology.organism_classification ,Molecular biology ,3. Good health ,chemistry ,Lentivirus ,HIV-1 ,Leukocytes, Mononuclear ,Parasitology ,Sequence Alignment ,030217 neurology & neurosurgery ,DNA ,Protein Binding - Abstract
SummaryThe expression of hundreds of interferon-stimulated genes (ISGs) causes the cellular “antiviral state” in which the replication of many viruses, including HIV-1, is attenuated. We conducted a screen for ISGs that inhibit HIV-1 virion production and found that 2′,3′-cyclic-nucleotide 3′-phosphodiesterase (CNP), a membrane-associated protein with unknown function in mammals has this property. CNP binds to the structural protein Gag and blocks HIV-1 particle assembly after Gag and viral RNA have associated with the plasma membrane. Several primate lentiviruses are CNP-sensitive, and CNP sensitivity/resistance is determined by a single, naturally dimorphic, codon (E/K40) in the matrix domain of Gag. Like other antiretroviral proteins, CNP displays interspecies variation in antiviral activity. Mice encode an inactive CNP variant and a single amino acid difference in murine versus human CNP determines Gag binding and antiviral activity. Some cell types express high levels of CNP and we speculate that CNP evolved to restrict lentivirus replication therein.
- Published
- 2012
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27. Cell-Based Screen Identifies Human Interferon-Stimulated Regulators of Listeria monocytogenes Infection
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Michael E. Abrams, Didi Chen, Sofya S. Perelman, Marcel Mettlen, Neal M. Alto, Jennifer L. Eitson, John W. Schoggins, and Alyssa Jimenez
- Subjects
0301 basic medicine ,RNA viruses ,medicine.disease_cause ,Pathology and Laboratory Medicine ,Polymerase Chain Reaction ,Biochemistry ,Immune Receptors ,Spectrum Analysis Techniques ,Interferon ,Animal Cells ,Medicine and Health Sciences ,Listeriosis ,Internalization ,lcsh:QH301-705.5 ,Pathogen ,media_common ,Connective Tissue Cells ,Immune System Proteins ,Virulence ,High-Throughput Nucleotide Sequencing ,Flow Cytometry ,3. Good health ,Bacterial Pathogens ,Antibody opsonization ,Medical Microbiology ,Connective Tissue ,Spectrophotometry ,Viral Pathogens ,Interferon Type I ,Viruses ,Cytophotometry ,Pathogens ,Cellular Types ,Anatomy ,medicine.drug ,Research Article ,Signal Transduction ,lcsh:Immunologic diseases. Allergy ,UNC93B1 ,media_common.quotation_subject ,Immunoblotting ,Immunology ,Context (language use) ,Biology ,Research and Analysis Methods ,Microbiology ,Cell Line ,03 medical and health sciences ,Immune system ,Listeria monocytogenes ,Gene Types ,Virology ,Retroviruses ,medicine ,Fc Receptors ,Genetics ,Humans ,Molecular Biology ,Microbial Pathogens ,030102 biochemistry & molecular biology ,Lentivirus ,Organisms ,Biology and Life Sciences ,Proteins ,Cell Biology ,Fibroblasts ,Listeria Monocytogenes ,030104 developmental biology ,Biological Tissue ,lcsh:Biology (General) ,Microscopy, Electron, Scanning ,Regulator Genes ,Parasitology ,Interferons ,lcsh:RC581-607 ,Transcriptome - Abstract
The type I interferon (IFN) activated transcriptional response is a critical antiviral defense mechanism, yet its role in bacterial pathogenesis remains less well characterized. Using an intracellular pathogen Listeria monocytogenes (Lm) as a model bacterial pathogen, we sought to identify the roles of individual interferon-stimulated genes (ISGs) in context of bacterial infection. Previously, IFN has been implicated in both restricting and promoting Lm growth and immune stimulatory functions in vivo. Here we adapted a gain-of-function flow cytometry based approach to screen a library of more than 350 human ISGs for inhibitors and enhancers of Lm infection. We identify 6 genes, including UNC93B1, MYD88, AQP9, and TRIM14 that potently inhibit Lm infection. These inhibitors act through both transcription-mediated (MYD88) and non-transcriptional mechanisms (TRIM14). Further, we identify and characterize the human high affinity immunoglobulin receptor FcγRIa as an enhancer of Lm internalization. Our results reveal that FcγRIa promotes Lm uptake in the absence of known host Lm internalization receptors (E-cadherin and c-Met) as well as bacterial surface internalins (InlA and InlB). Additionally, FcγRIa-mediated uptake occurs independently of Lm opsonization or canonical FcγRIa signaling. Finally, we established the contribution of FcγRIa to Lm infection in phagocytic cells, thus potentially linking the IFN response to a novel bacterial uptake pathway. Together, these studies provide an experimental and conceptual basis for deciphering the role of IFN in bacterial defense and virulence at single-gene resolution., Author Summary While the type I interferon response is known to be activated by both viruses and bacteria, it has mostly been characterized in terms of its antiviral properties. Listeria monocytogenes, an opportunistic Gram-positive bacterial pathogen with up to 50% mortality rate and a variety of clinical manifestations, is a potent activator of interferon secretion. In mouse models, interferon has been previously implicated in both restricting and promoting L. monocytogenes infection. Here, we utilized a high-throughput flow-cytometry based approach to screen a library of human interferon I stimulated genes (ISGs) and identified regulators of L. monocytogenes infection. These include inhibitors that act through both transcriptional (MYD88) and transcription-independent (TRIM14) mechanisms. Strikingly, expression of the human high affinity immunoglobulin receptor FcγRIa (CD64) was found to potently enhance L. monocytogenes infection. Both biochemical and cellular studies indicate that FcγRIa increases primary invasion of L. monocytogenes through a previously uncharacterized IgG-independent internalization mechanism. Together, these studies provide an important insight into the complex role of interferon response in bacterial virulence and host defense.
- Published
- 2016
28. NLRX1 Helps HIV Avoid a STING Operation
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John W. Schoggins and Katrina B. Mar
- Subjects
0301 basic medicine ,Innate immune system ,Human immunodeficiency virus (HIV) ,Acute infection ,Biology ,Evasion (ethics) ,medicine.disease_cause ,Sting operation ,Microbiology ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Immune system ,Virology ,Immunology ,medicine ,Parasitology ,Latency (engineering) ,NLRX1 ,030215 immunology - Abstract
Immune evasion by HIV-1 during acute infection is critical for the establishment of latency. Barouch et al. (2016) and Guo et al. (2016) demonstrate in independent studies that Nod-like receptor X1 (NLRX1) may facilitate early systemic dissemination of HIV-1 by inhibiting the virus-triggered innate immune response.
- Published
- 2016
29. Interferon-stimulated genes and their antiviral effector functions
- Author
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John W. Schoggins and Charles M. Rice
- Subjects
Biology ,Article ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Viral life cycle ,Transcription (biology) ,Interferon ,Virology ,medicine ,Animals ,Humans ,Gene ,030304 developmental biology ,Regulation of gene expression ,0303 health sciences ,Effector ,Gene Expression Profiling ,virus diseases ,Immunity, Innate ,3. Good health ,Gene Expression Regulation ,Viral replication ,Virus Diseases ,030220 oncology & carcinogenesis ,Interferon Type I ,Viruses ,Interferon type I ,medicine.drug - Abstract
Many viruses trigger the type I interferon (IFN) system, leading to the transcription of hundreds of interferon-stimulated genes (ISGs). The products of these ISGs exert numerous antiviral effector functions, many of which are still not fully described. Recent efforts have been aimed at identifying which ISGs are antiviral and further characterizing their mechanisms of action. IFN effectors vary widely in their magnitude of inhibitory activity and display combinatorial antiviral properties. Collectively, ISGs can target almost any step in a virus life cycle. Some of the most potent antiviral effectors reinforce the system by further inducing IFN or ISGs. Other genes enhance or facilitate viral replication, suggesting that some viruses may have evolved to co-opt IFN effectors for a survival advantage.
- Published
- 2011
30. A diverse range of gene products are effectors of the type I interferon antiviral response
- Author
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Maryline Panis, Charles M. Rice, Mary Murphy, Christopher T. Jones, Sam J. Wilson, Paul D. Bieniasz, and John W. Schoggins
- Subjects
viruses ,Biology ,Virus Replication ,medicine.disease_cause ,Article ,Virus ,Cell Line ,Interferon ,medicine ,Animals ,Humans ,Genetics ,Multidisciplinary ,Cyclic GMP-AMP synthase ,Gene Expression Profiling ,Interferon-stimulated gene ,virus diseases ,MDA5 ,Virology ,HEK293 Cells ,IRF1 ,Gene Expression Regulation ,Viral replication ,Protein Biosynthesis ,Interferon Type I ,Viruses ,Venezuelan equine encephalitis virus ,medicine.drug - Abstract
The type I interferon response protects cells against invading viral pathogens. The cellular factors that mediate this defence are the products of interferon-stimulated genes (ISGs). Although hundreds of ISGs have been identified since their discovery more than 25 years ago, only a few have been characterized with respect to antiviral activity. For most ISG products, little is known about their antiviral potential, their target specificity and their mechanisms of action. Using an overexpression screening approach, here we show that different viruses are targeted by unique sets of ISGs. We find that each viral species is susceptible to multiple antiviral genes, which together encompass a range of inhibitory activities. To conduct the screen, more than 380 human ISGs were tested for their ability to inhibit the replication of several important human and animal viruses, including hepatitis C virus, yellow fever virus, West Nile virus, chikungunya virus, Venezuelan equine encephalitis virus and human immunodeficiency virus type-1. Broadly acting effectors included IRF1, C6orf150 (also known as MB21D1), HPSE, RIG-I (also known as DDX58), MDA5 (also known as IFIH1) and IFITM3, whereas more targeted antiviral specificity was observed with DDX60, IFI44L, IFI6, IFITM2, MAP3K14, MOV10, NAMPT (also known as PBEF1), OASL, RTP4, TREX1 and UNC84B (also known as SUN2). Combined expression of pairs of ISGs showed additive antiviral effects similar to those of moderate type I interferon doses. Mechanistic studies uncovered a common theme of translational inhibition for numerous effectors. Several ISGs, including ADAR, FAM46C, LY6E and MCOLN2, enhanced the replication of certain viruses, highlighting another layer of complexity in the highly pleiotropic type I interferon system.
- Published
- 2011
31. Splicing Diversity of the Human OCLN Gene and Its Biological Significance for Hepatitis C Virus Entry
- Author
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Evgenia Korol, Ludmila Prokunina-Olsson, Charles M. Rice, Kathy Mu, Thomas R. O'Brien, Indu Kohaar, Alexander Ploss, and John W. Schoggins
- Subjects
Hepatitis C virus ,Molecular Sequence Data ,Immunology ,Hepacivirus ,Biology ,medicine.disease_cause ,Occludin ,Microbiology ,Virus ,Exon ,Virology ,medicine ,Humans ,Protein Isoforms ,Tissue Distribution ,Gene ,Chromosomes, Human, Pair 15 ,Liver Neoplasms ,Alternative splicing ,Membrane Proteins ,Exons ,Hepatitis C, Chronic ,Virus Internalization ,Virus-Cell Interactions ,Alternative Splicing ,Liver ,Insect Science ,RNA splicing ,Tissue tropism ,HeLa Cells - Abstract
Persistent hepatitis C virus (HCV) infection is a primary etiological factor for the development of chronic liver disease, including cirrhosis and cancer. A recent study identified occludin (OCLN), an integral tight junction protein, as one of the key factors for HCV entry into cells. We explored the splicing diversity of OCLN in normal human liver and observed variable expression of alternative splice variants, including two known forms ( WT-OCLN and OCLN-ex4del ) and six novel forms ( OCLN-ex7ext , OCLN-ex3pdel , OCLN-ex3del , OCLN-ex3 - 4del , OCLN-ex3p-9pdel , and OCLN-ex3p-7pdel ). Recombinant protein isoforms WT-OCLN and OCLN-ex7ext, which retained the HCV-interacting MARVEL domain, were expressed on the cell membrane and were permissive for HCV infection in in vitro infectivity assays. All other forms lacked the MARVEL domain, were expressed in the cytoplasm, and were nonpermissive for HCV infection. Additionally, we observed variable expression of OCLN splicing forms across human tissues and cell lines. Our study suggests that the remarkable natural splicing diversity of OCLN might contribute to HCV tissue tropism and possibly modify the outcome of HCV infection in humans. Genetic factors crucial for regulation of OCLN expression and susceptibility to HCV infection remain to be elucidated.
- Published
- 2010
32. Fiber and Penton Base Capsid Modifications Yield Diminished Adenovirus Type 5 Transduction and Proinflammatory Gene Expression with Retention of Antigen-Specific Humoral Immunity
- Author
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John W. Schoggins and Erik Falck-Pedersen
- Subjects
Adenoviridae Infections ,viruses ,Genetic Vectors ,Immunology ,Gene Expression ,Antibodies, Viral ,medicine.disease_cause ,Microbiology ,Adenoviridae ,Cell Line ,Mice ,Transduction (genetics) ,Immune system ,Transduction, Genetic ,Virology ,medicine ,Animals ,Humans ,Antigens, Viral ,RGD motif ,biology ,Macrophage Activation ,biology.organism_classification ,Molecular biology ,Mastadenovirus ,Capsid ,Cell culture ,Insect Science ,Humoral immunity ,Cytokines ,Pathogenesis and Immunity ,Capsid Proteins ,Chemokines - Abstract
Fiber and penton base capsid proteins of adenovirus type 5 (Ad5) mediate a well-characterized two-step entry pathway in permissive tissue culture cell lines. Fiber binds with high affinity to the cell surface coxsackievirus-and-adenovirus receptor (CAR), and penton base facilitates viral internalization by binding αv integrins through an RGD motif. In vivo, the entry pathway is complicated by interactions of capsid proteins with additional cell surface molecules and blood factors. When administered systemically in mice, adenovirus vectors (Adv) localize primarily to hepatic tissue, resulting in efficient gene transduction and potent activation of the host antiviral immune response. The goal of the present study was to detarget Adv uptake through fiber and penton base capsid protein manipulations and determine how detargeted vectors influence transduction efficiency, inflammatory activation, and activation of the adaptive arm of the immune system. By manipulating fiber and the penton base, we have generated highly detargeted vectors (up to 1,200-fold reduction in transgene expression in vivo) with reduced macrophage stimulatory activity in vitro and in vivo. In spite of the diminished transduction and macrophage activation, the detargeted vectors induce strong neutralizing immunity as well as efficient antitransgene antibody. Three of the modified vectors produce antitransgene humoral immunity at levels that exceed or are equal to that seen with an unmodified Ad5-based vector. The fiber-pseudotyped and penton base constructs with RGD deleted have attributes that could be important enhancements in a number of vaccine applications.
- Published
- 2006
33. Influence of Fiber Detargeting on Adenovirus-Mediated Innate and Adaptive Immune Activation
- Author
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John W. Schoggins, Nicola J. Philpott, Marcelo Nociari, and Erik Falck-Pedersen
- Subjects
Coxsackie and Adenovirus Receptor-Like Membrane Protein ,Virus genetics ,Chemokine ,Genetic Vectors ,Immunology ,Biology ,medicine.disease_cause ,Microbiology ,Adenoviridae ,Cell Line ,Mice ,Transduction (genetics) ,Transduction, Genetic ,Virology ,medicine ,Animals ,Humans ,Vector (molecular biology) ,Antigen-presenting cell ,Antigens, Viral ,Macrophages ,Dendritic Cells ,Acquired immune system ,Adaptation, Physiological ,Molecular biology ,Immunity, Innate ,Virus-Cell Interactions ,Insect Science ,DNA, Viral ,Hepatocytes ,Pseudotyping ,biology.protein ,Cytokines ,Receptors, Virus ,Capsid Proteins ,Female ,Chemokines - Abstract
The major adenovirus (Ad) capsid proteins hexon, penton, and fiber influence the efficiency and tropism of gene transduction by Ad vectors. Fiber is the high-affinity receptor binding protein that serves to mediate cell attachment in vitro when using coxsackie-adenovirus receptor (CAR)-containing cell lines. This contrasts with transduction efficiency in macrophages or dendritic cells that lack high concentrations of CAR. To determine how fiber influences gene transduction and immune activation in a murine model, we have characterized Ad type 5 (Ad5) vectors with two classes of chimeric fiber, CAR binding and non-CAR binding. In a systemic infection, Ad5 fiber contributes to DNA localization and vector transduction in hepatic tissue. However, the majority of vector localization is due to Ad5 fiber-specific functions distinct from CAR binding. CAR-directed transduction occurs but at a modest level. In contrast to CAR binding vectors, the F7 and F7F41S non-CAR-binding vectors demonstrate a 2-log decrease in hepatic transduction, with a 10-fold decrease in the amount of vector DNA localizing to the hepatic tissue. To characterize the innate response to early infection using fiber chimeric vectors, intrahepatic cytokine and chemokine mRNAs were quantified 5 hours postinfection. Tumor necrosis factor alpha mRNA levels resulting from Ad5 fiber infections were elevated compared to viruses expressing serotype 7 or 41 fiber. Levels of chemokine mRNA (gamma interferon-inducible protein 10, T-cell activation gene 3, and macrophage inflammatory protein 1β) were 10- to 20-fold higher with CAR binding vectors (Ad5 and F41T) than with non-CAR-binding vectors (F7 and F7F41S). In spite of quantitative differences in vector localization and innate activation, fiber pseudotyping did not significantly change the outcome of anti-Ad adaptive immunity. All vectors were cleared with the same kinetics as wild-type Ad5 vectors, and each induced neutralizing antibody. Although non-CAR-binding vectors were impaired in transduction by nearly 2 orders of magnitude, the level of antitransgene immunity was the same for each of the vectors. Using primary bone marrow-derived macrophages and dendritic cells, we demonstrate that transduction, induction of cytokine/chemokine, and phenotypic maturation of these antigen-presenting cells are independent of fiber content. Our data support a model where fiber-mediated hepatic localization enhances innate responses to virus infection but minimally impacts on adaptive immunity.
- Published
- 2005
34. Pan-viral specificity of IFN-induced genes reveals new roles for cGAS in innate immunity
- Author
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Herbert W. Virgin, Alan Aderem, Eric J. Snijder, Alexander V. Ratushny, Vladimir Litvak, John W. Schoggins, Maria D. Gainey, Katrina B. Mar, Naoko Imanaka, John D. Aitchison, Jennifer L. Eitson, R. Blake Richardson, Richard M. Elliott, Balaji Manicassamy, Vincent R. Racaniello, Rea Dabelic, Donna A. MacDuff, Adolfo García-Sastre, Charles M. Rice, Bimmi Shrestha, Wayne M. Yokoyama, and Michael S. Diamond
- Subjects
viruses ,Biology ,Virus ,Article ,Substrate Specificity ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,0302 clinical medicine ,Interferon ,medicine ,Animals ,Cluster Analysis ,RNA Viruses ,Gene ,Virus classification ,030304 developmental biology ,Gene Library ,Genetics ,Mice, Knockout ,0303 health sciences ,Multidisciplinary ,Cyclic GMP-AMP synthase ,DNA Viruses ,RNA ,Membrane Proteins ,RNA virus ,biology.organism_classification ,Flow Cytometry ,Virology ,Nucleotidyltransferases ,Immunity, Innate ,3. Good health ,STAT1 Transcription Factor ,chemistry ,030220 oncology & carcinogenesis ,Viruses ,Interferon Regulatory Factor-3 ,Interferons ,DNA ,medicine.drug - Abstract
The type I interferon (IFN) response protects cells from viral infection by inducing hundreds of interferon-stimulated genes (ISGs), some of which encode direct antiviral effectors. Recent screening studies have begun to catalogue ISGs with antiviral activity against several RNA and DNA viruses. However, antiviral ISG specificity across multiple distinct classes of viruses remains largely unexplored. Here we used an ectopic expression assay to screen a library of more than 350 human ISGs for effects on 14 viruses representing 7 families and 11 genera. We show that 47 genes inhibit one or more viruses, and 25 genes enhance virus infectivity. Comparative analysis reveals that the screened ISGs target positive-sense single-stranded RNA viruses more effectively than negative-sense single-stranded RNA viruses. Gene clustering highlights the cytosolic DNA sensor cyclic GMP-AMP synthase (cGAS, also known as MB21D1) as a gene whose expression also broadly inhibits several RNA viruses. In vitro, lentiviral delivery of enzymatically active cGAS triggers a STING-dependent, IRF3-mediated antiviral program that functions independently of canonical IFN/STAT1 signalling. In vivo, genetic ablation of murine cGAS reveals its requirement in the antiviral response to two DNA viruses, and an unappreciated contribution to the innate control of an RNA virus. These studies uncover new paradigms for the preferential specificity of IFN-mediated antiviral pathways spanning several virus families.
- Published
- 2014
35. Lipids in innate antiviral defense
- Author
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John W. Schoggins and Glenn Randall
- Subjects
Cancer Research ,Membrane lipids ,Lipid composition ,viruses ,Virus diseases ,Biology ,Microbiology ,Article ,03 medical and health sciences ,Membrane Lipids ,Viral envelope ,Immunity ,Immunology and Microbiology(all) ,Virology ,Animals ,Humans ,Molecular Biology ,030304 developmental biology ,0303 health sciences ,Innate immune system ,030306 microbiology ,Extramural ,Virus Internalization ,Immunity, Innate ,3. Good health ,Cell biology ,Membrane ,Virus Diseases ,Viruses ,Parasitology - Abstract
It is becoming apparent that infections by a major class of viruses, those with envelopes, can be inhibited during their entry at the step of fusion with cellular membranes. In this review, we discuss multiple innate immune mechanisms that have evolved to modify the lipid composition of cellular and viral membranes to inhibit virion fusion of enveloped viruses.
- Published
- 2013
36. Viruses carry antiviral cargo
- Author
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John W. Schoggins
- Subjects
Immune defense ,Multidisciplinary ,biology ,Interferon beta ,Host (biology) ,Arms race ,Bridgeman ,biology.organism_classification ,Virology ,Viral infection ,chemistry.chemical_compound ,Muromegalovirus ,chemistry ,Vaccinia - Abstract
Infected cells generate a factor that is incorporated into viruses and transferred to other cells [Also see Reports by Bridgeman et al. and Gentili et al. ]
- Published
- 2015
37. Host restriction factor screening: let the virus do the work
- Author
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John W. Schoggins and Michael S. Diamond
- Subjects
Genetics ,Sindbis virus ,Cancer Research ,biology ,viruses ,RNA ,biology.organism_classification ,Virus Replication ,Microbiology ,Virology ,Virus ,Viral replication ,Viral evolution ,Immunology and Microbiology(all) ,Host-Pathogen Interactions ,Animals ,Humans ,Parasitology ,Genetic Testing ,Sindbis Virus ,Permissive ,Gene ,Molecular Biology ,Oncovirus - Abstract
In this issue of Cell Host & Microbe, Varble et al. (2013) engineer a library of RNA viruses to express small interfering RNAs and couple this with the power of virus evolution and selection to screen for host genes that when silenced resulted in greater viral infection in vivo.
- Published
- 2013
38. Dengue reporter viruses reveal viral dynamics in interferon receptor-deficient mice and sensitivity to interferon effectors in vitro
- Author
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Michael Feulner, Alexander Ploss, Naoko Imanaka, Charles M. Rice, John W. Schoggins, Marcus Dorner, and Mary Murphy
- Subjects
viruses ,Green Fluorescent Proteins ,Dengue virus ,In Vitro Techniques ,medicine.disease_cause ,Virus ,Dengue ,Mice ,Interferon ,Genes, Reporter ,Luciferases, Firefly ,Chlorocebus aethiops ,medicine ,Bioluminescence imaging ,Animals ,Luciferase ,Vero Cells ,Gene Library ,Analysis of Variance ,Multidisciplinary ,biology ,Interferon-stimulated gene ,virus diseases ,biochemical phenomena, metabolism, and nutrition ,Dengue Virus ,Biological Sciences ,biology.organism_classification ,Flow Cytometry ,Virology ,High-Throughput Screening Assays ,Flavivirus ,Vero cell ,Genetic Engineering ,medicine.drug ,Plasmids - Abstract
Dengue virus (DENV) is a global disease threat for which there are no approved antivirals or vaccines. Establishing state-of-the-art screening systems that rely on fluorescent or luminescent reporters may accelerate the development of anti-DENV therapeutics. However, relatively few reporter DENV platforms exist. Here, we show that DENV can be genetically engineered to express a green fluorescent protein or firefly luciferase. Reporter viruses are infectious in vitro and in vivo and are sensitive to antiviral compounds, neutralizing antibodies, and interferons. Bioluminescence imaging was used to follow the dynamics of DENV infection in mice and revealed that the virus localized predominantly to lymphoid and gut-associated tissues. The high-throughput potential of reporter DENV was demonstrated by screening a library of more than 350 IFN-stimulated genes for antiviral activity. Several antiviral effectors were identified, and they targeted DENV at two distinct life cycle steps. These viruses provide a powerful platform for applications ranging from validation of vaccine candidates to antiviral discovery.
- Published
- 2012
39. Multiple interferon stimulated genes synergize with the zinc finger antiviral protein to mediate anti-alphavirus activity
- Author
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Margaret R. MacDonald, Charles M. Rice, Sophiya Karki, John W. Schoggins, Suyan Tian, Melody M. H. Li, and Lee, Young-Min
- Subjects
RNA viruses ,Anatomy and Physiology ,lcsh:Medicine ,Viral classification ,Interferon ,Immune Physiology ,Cricetinae ,lcsh:Science ,Host factor ,0303 health sciences ,Multidisciplinary ,030302 biochemistry & molecular biology ,virus diseases ,RNA-Binding Proteins ,hemic and immune systems ,Drug Synergism ,Innate Immunity ,3. Good health ,Infectious Diseases ,Gene Knockdown Techniques ,Cytokines ,Medicine ,Research Article ,medicine.drug ,Sindbis virus ,Interferon Regulatory Factor 2 ,General Science & Technology ,Mechanisms of Resistance and Susceptibility ,1.1 Normal biological development and functioning ,Immunology ,Alpha interferon ,chemical and pharmacologic phenomena ,Alphavirus ,Biology ,Microbiology ,Cell Line ,Vaccine Related ,03 medical and health sciences ,Underpinning research ,Virology ,Biodefense ,medicine ,Genetics ,Animals ,Humans ,030304 developmental biology ,Prevention ,lcsh:R ,Immunity ,Interferon-alpha ,biology.organism_classification ,Rats ,Vector-Borne Diseases ,Emerging Infectious Diseases ,Viral replication ,Immune System ,Togaviridae ,Clinical Immunology ,lcsh:Q ,Sindbis Virus ,Carrier Proteins ,Interferon Regulatory Factor-2 - Abstract
The zinc finger antiviral protein (ZAP) is a host factor that mediates inhibition of viruses in the Filoviridae, Retroviridae and Togaviridae families. We previously demonstrated that ZAP blocks replication of Sindbis virus (SINV), the prototype Alphavirus in the Togaviridae family at an early step prior to translation of the incoming genome and that synergy between ZAP and one or more interferon stimulated genes (ISGs) resulted in maximal inhibitory activity. The present study aimed to identify those ISGs that synergize with ZAP to mediate Alphavirus inhibition. Using a library of lentiviruses individually expressing more than 350 ISGs, we screened for inhibitory activity in interferon defective cells with or without ZAP overexpression. Confirmatory tests of the 23 ISGs demonstrating the largest infection reduction in combination with ZAP revealed that 16 were synergistic. Confirmatory tests of all potentially synergistic ISGs revealed 15 additional ISGs with a statistically significant synergistic effect in combination with ZAP. These 31 ISGs are candidates for further mechanistic studies. The number and diversity of the identified ZAP-synergistic ISGs lead us to speculate that ZAP may play an important role in priming the cell for optimal ISG function.
- Published
- 2012
40. A phospholipase linkAGE to SARS susceptibility
- Author
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John W. Schoggins
- Subjects
Immunology ,Age Factors ,Dendritic Cells ,News ,Biology ,Phospholipase ,Severe Acute Respiratory Syndrome ,Insights ,Group II Phospholipases A2 ,Lipids ,Virology ,Immunity, Innate ,CD11c Antigen ,Mice, Inbred C57BL ,Mice ,Oxidative Stress ,Disease susceptibility ,Animals ,Humans ,Immunology and Allergy ,Disease Susceptibility ,Lung - Abstract
Oxidative stress and chronic low-grade inflammation in the lungs are associated with aging and may contribute to age-related immune dysfunction. To maintain lung homeostasis, chronic inflammation is countered by enhanced expression of proresolving/antiinflammatory factors. Here, we show that age-dependent increases of one such factor in the lungs, a phospholipase A2 (PLA2) group IID (PLA2G2D) with antiinflammatory properties, contributed to worse outcomes in mice infected with severe acute respiratory syndrome-coronavirus (SARS-CoV). Strikingly, infection of mice lacking PLA2G2D expression (Pla2g2d(-/-) mice) converted a uniformly lethal infection to a nonlethal one (80% survival), subsequent to development of enhanced respiratory DC migration to the draining lymph nodes, augmented antivirus T cell responses, and diminished lung damage. We also observed similar effects in influenza A virus-infected middle-aged Pla2g2d(-/-) mice. Furthermore, oxidative stress, probably via lipid peroxidation, was found to induce PLA2G2D expression in mice and in human monocyte-derived macrophages. Thus, our results suggest that directed inhibition of a single inducible phospholipase, PLA2G2D, in the lungs of older patients with severe respiratory infections is potentially an attractive therapeutic intervention to restore immune function.
- Published
- 2015
- Full Text
- View/download PDF
41. A genetically humanized mouse model for hepatitis C virus infection
- Author
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Alexander Ploss, Charles M. Rice, Marcus Dorner, Qian Feng, Kathy Mu, Joshua A. Horwitz, Mansun Law, John W. Schoggins, Dennis R. Burton, Walter T. Barry, Maria Teresa Catanese, Justin B. Robbins, and Christopher T. Jones
- Subjects
Viral pathogenesis ,Hepacivirus ,medicine.disease_cause ,Recombinant virus ,Antigens, CD81 ,Mice ,0302 clinical medicine ,CD81 ,Claudin-1 ,Orthopoxvirus ,Cells, Cultured ,0303 health sciences ,Multidisciplinary ,biology ,Scavenger Receptors, Class B ,Hepatitis C ,3. Good health ,Science & Technology - Other Topics ,Receptors, Virus ,030211 gastroenterology & hepatology ,FIBROBLASTS ,Genotype ,General Science & Technology ,Hepatitis C virus ,Transfection ,Virus ,Article ,Tetraspanin 28 ,Adenoviridae ,03 medical and health sciences ,Viral entry ,Antigens, CD ,HUMAN LIVER ,MD Multidisciplinary ,medicine ,Animals ,Humans ,Antibodies, Blocking ,030304 developmental biology ,CELL-CULTURE ,Science & Technology ,RECEPTOR ,MULTIDISCIPLINARY SCIENCES ,Immunization, Passive ,Membrane Proteins ,Pathogenesis and modulation of inflammation Infection and autoimmunity [N4i 1] ,biology.organism_classification ,Virology ,Disease Models, Animal ,Viral Tropism ,B TYPE-I ,Humanized mouse ,Immunology ,REPLICATION ,ANTIBODIES ,Tissue tropism ,Hepatocytes ,HIGH-DENSITY-LIPOPROTEIN - Abstract
Item does not contain fulltext Hepatitis C virus (HCV) remains a major medical problem. Antiviral treatment is only partially effective and a vaccine does not exist. Development of more effective therapies has been hampered by the lack of a suitable small animal model. Although xenotransplantation of immunodeficient mice with human hepatocytes has shown promise, these models are subject to important challenges. Building on the previous observation that CD81 and occludin comprise the minimal human factors required to render mouse cells permissive to HCV entry in vitro, we attempted murine humanization via a genetic approach. Here we show that expression of two human genes is sufficient to allow HCV infection of fully immunocompetent inbred mice. We establish a precedent for applying mouse genetics to dissect viral entry and validate the role of scavenger receptor type B class I for HCV uptake. We demonstrate that HCV can be blocked by passive immunization, as well as showing that a recombinant vaccinia virus vector induces humoral immunity and confers partial protection against heterologous challenge. This system recapitulates a portion of the HCV life cycle in an immunocompetent rodent for the first time, opening opportunities for studying viral pathogenesis and immunity and comprising an effective platform for testing HCV entry inhibitors in vivo.
- Published
- 2011
42. Expression of paramyxovirus V proteins promotes replication and spread of hepatitis C virus in cultures of primary human fetal liver cells
- Author
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Timothy P. Sheahan, Svetlana Marukian, Charles M. Rice, Walter T. Barry, Christopher T. Jones, Alexander Ploss, Kartik Trehan, Lynn B. Dustin, John W. Schoggins, Sangeeta N. Bhatia, Maria Teresa Catanese, Linda Andrus, Institute for Medical Engineering and Science, Harvard University--MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science, Trehan, Kartik, and Bhatia, Sangeeta N.
- Subjects
Viral Hepatitis ,Hepacivirus ,Hepatitis C virus ,Primary Cell Culture ,Population ,Virus Replication ,medicine.disease_cause ,Virus ,Measles virus ,Viral Proteins ,03 medical and health sciences ,0302 clinical medicine ,Transduction, Genetic ,Interferon ,medicine ,Humans ,education ,030304 developmental biology ,0303 health sciences ,education.field_of_study ,Hepatology ,biology ,Interleukins ,Lentivirus ,virus diseases ,biology.organism_classification ,Hepatitis C ,Virology ,3. Good health ,Liver ,Viral replication ,Cell culture ,030220 oncology & carcinogenesis ,Hepatocytes ,Interferons ,medicine.drug - Abstract
Here we demonstrate that primary cultures of human fetal liver cells (HFLC) reliably support infection with laboratory strains of hepatitis C virus (HCV), although levels of virus replication vary significantly between different donor cell preparations and frequently decline in a manner suggestive of active viral clearance. To investigate possible contributions of the interferon (IFN) system to control HCV infection in HFLC, we exploited the well-characterized ability of paramyxovirus (PMV) V proteins to counteract both IFN induction and antiviral signaling. The V proteins of measles virus (MV) and parainfluenza virus 5 (PIV5) were introduced into HFLC using lentiviral vectors encoding a fluorescent reporter for visualization of HCV-infected cells. V protein-transduced HFLC supported enhanced (10 to 100-fold) levels of HCV infection relative to untransduced or control vector-transduced HFLC. Infection was assessed by measurement of virus-driven luciferase, by assays for infectious HCV and viral RNA, and by direct visualization of HCV-infected hepatocytes. Live cell imaging between 48 and 119 hours postinfection demonstrated little or no spread of infection in the absence of PMV V protein expression. In contrast, V protein-transduced HFLC showed numerous HCV infection events. V protein expression efficiently antagonized the HCV-inhibitory effects of added IFNs in HFLC. In addition, induction of the type III IFN, IL29, following acute HCV infection was inhibited in V protein-transduced cultures. Conclusion: These studies suggest that the cellular IFN response plays a significant role in limiting the spread of HCV infection in primary hepatocyte cultures. Strategies aimed at dampening this response may be key to further development of robust HCV culture systems, enabling studies of virus pathogenicity and the mechanisms by which HCV spreads in its natural host cell population., National Institutes of Health (U.S.) (NIH Roadmap for Medical Research Grant 1 R01 DK085713-01), Greenberg Institute for Medical Research, Starr Foundation
- Published
- 2011
43. Real-time imaging of hepatitis C virus infection using a fluorescent cell-based reporter system
- Author
-
Andrew J. Syder, John W. Schoggins, Salman R. Khetani, Charles M. Rice, Margaret R. MacDonald, Christopher T. Jones, Lok Man J. Law, Maria Teresa Catanese, Sangeeta N. Bhatia, Alexander Ploss, Thomas S. Oh, Harvard University--MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science, Khetani, Salman R., and Bhatia, Sangeeta N.
- Subjects
Hepacivirus ,medicine.medical_treatment ,Hepatitis C virus ,Cell ,Population ,Biomedical Engineering ,Bioengineering ,Liver transplantation ,medicine.disease_cause ,Applied Microbiology and Biotechnology ,03 medical and health sciences ,0302 clinical medicine ,Fluorescent cell ,Computer Systems ,Genes, Reporter ,medicine ,education ,Gene ,030304 developmental biology ,0303 health sciences ,education.field_of_study ,biology ,Staining and Labeling ,biology.organism_classification ,Image Enhancement ,Virology ,3. Good health ,Genetically modified organism ,medicine.anatomical_structure ,Microscopy, Fluorescence ,030220 oncology & carcinogenesis ,Molecular Medicine ,Biotechnology - Abstract
Author Manuscript 2010 August 1, Hepatitis C virus (HCV), which infects 2–3% of the world population, is a causative agent of chronic hepatitis and the leading indication for liver transplantation1. The ability to propagate HCV in cell culture (HCVcc) is a relatively recent breakthrough and a key tool in the quest for specific antiviral therapeutics. Monitoring HCV infection in culture generally involves bulk population assays, use of genetically modified viruses and/or terminal processing of potentially precious samples. Here we develop a cell-based fluorescent reporter system that allows sensitive distinction of individual HCV-infected cells in live or fixed samples. We demonstrate use of this technology for several previously intractable applications, including live-cell imaging of viral propagation and host response, as well as visualizing infection of primary hepatocyte cultures. Integration of this reporter with modern image-based analysis methods could open new doors for HCV research., New York (State). Dept. of Health (Empire State Stem Cell Fund Contract C023046), United States. Public Health Service (Grant R01 DK56966), National Institutes of Health (U.S.) (Roadmap for Medical Research Grant 1 R01 DK085713-01), Howard Hughes Medical Institute (Investigator)
- Published
- 2009
44. Serotype 5 Adenovirus fiber (F7F41S) chimeric vectors incur packaging deficiencies when targeting peptides are inserted into Ad41 short fiber
- Author
-
Erik Falck-Pedersen and John W. Schoggins
- Subjects
viruses ,Genetic Vectors ,Molecular Sequence Data ,Sequence alignment ,Biology ,Virus ,Article ,Viral vector ,Cell Line ,Transduction (genetics) ,Viral Proteins ,Viral entry ,Virology ,Adenovirus ,Animals ,Humans ,Fiber ,Amino Acid Sequence ,Gene ,Peptide sequence ,Adenoviruses, Human ,Virus Assembly ,Gene Transfer Techniques ,Cell biology ,Protein Structure, Tertiary ,Capsid structure ,Mutagenesis, Insertional ,Capsid ,Capsid Proteins ,Sequence Alignment - Abstract
Adenovirus is a well-established viral gene transfer model system that presents two major hurdles when being considered for cell-specific targeting applications. First is the need to detarget the vector from inherent host binding mechanisms, and second is the need to establish a productive and stable method to retarget the vector to a desired cell receptor. In previous studies we had generated an adenovirus vector platform that lacks the normal targeting attributes derived from the fiber and penton capsid proteins. In the current study we characterized our detargeted Ad5-based vectors (Ad5.F7F41S and Ad5.F7F41SΔRGD) as platforms for novel retargeted viruses. The experimental strategy relied on incorporating small peptide ligands into several sites of the Ad 41short fiber knob domain (AB, CD, HI, G and Cterm). Reengineering of Ad41 short fiber resulted either in a bypass to fiber 7 usage, or in a dominant negative packaging/production deficiency phenotype. Under specific growth conditions we could remedy some of the capsid deficiencies and generate high titer viruses. However when examined by Western blot analysis, the resulting viruses were still defective in capsid content. The tandem fiber F7F41S platform has revealed an unanticipated sensitivity of Adenovirus packaging to fiber 41short structural modifications. These studies indicate fiber assembly into an intact virion or fiber influenced capsid stability as a bottleneck to efficient particle production. We also demonstrate that virus particles characterized as mature virions following CsCl banding can vary significantly in capsid protein content. Considering the complexity of virus entry into a target cell, modified “mature virions” may be compromised at the level of transduction not only through the intended modification, but also by virtue of secondary structural packaging conflicts.
- Published
- 2009
45. Adenovirus Capsid Chimeras: Fiber Terminal Exon Insertions/Gene Replacements in the Major Late Transcription Unit
- Author
-
Erik Falck-Pedersen, John W. Schoggins, and Jason G. D. Gall
- Subjects
Immune recognition ,Messenger RNA ,Exon ,Immune system ,Capsid ,Transcription (biology) ,viruses ,medicine ,Adenovirus infection ,Biology ,medicine.disease ,Virology ,Gene - Abstract
The adenovirus major late transcription unit (MLTU) encodes the main structural capsid proteins. Expression from the MLTU is accomplished through alternative mRNA processing and use of a terminal exon coding strategy. The capsid proteins hexon, penton, and fiber contribute to efficient infection by adenovirus, and each contributes in some manner to the antiviral immune response against adenovirus infection. The ability to manipulate these genes affords one the opportunity to "detarget" adenovirus, to retarget adenovirus, and to alter immune recognition. In this chapter, we are presenting a terminal exon-replacement strategy that can be used to genetically manipulate capsid proteins expressed from the MLTU. An emphasis will be placed on manipulations of fiber as an intact terminal exon.
- Published
- 2007
46. Sensing Infection by Adenovirus: Toll-Like Receptor-Independent Viral DNA Recognition Signals Activation of the Interferon Regulatory Factor 3 Master Regulator▿
- Author
-
John W. Schoggins, Oksana Ocheretina, Marcelo Nociari, and Erik Falck-Pedersen
- Subjects
Male ,viruses ,Adenoviridae Infections ,Immunology ,Alpha interferon ,Cellular Response to Infection ,Antigen-Presenting Cells ,Biology ,medicine.disease_cause ,Microbiology ,Mice ,Viral entry ,Interferon ,Virology ,medicine ,Animals ,Phosphorylation ,Cells, Cultured ,Toll-like receptor ,Interleukin-6 ,Tumor Necrosis Factor-alpha ,Interferon-alpha ,Fibroblasts ,Acquired immune system ,Adenoviridae ,Mice, Inbred C57BL ,Disease Models, Animal ,Insect Science ,DNA, Viral ,Female ,Interferon Regulatory Factor-3 ,IRF3 ,medicine.drug ,Interferon regulatory factors ,Signal Transduction - Abstract
Infection with adenovirus vectors (AdV) results in rapid activation of innate immunity, which serves the dual purpose of stimulating inflammatory antiviral host defenses and the adaptive immune system. Viral recognition by macrophages, dendritic cells, and other cell types requires an ability to sense the presence of a foreign molecular pattern by “pattern recognition receptors.” The nature of the adenoviral sensor, the target ligand of the sensor, and the downstream antiviral signaling response triggered by virus infection have not been defined for this nonenveloped double-stranded DNA (dsDNA) virus. We have identified four critical links involved in AdV recognition by murine antigen-presenting cells (APC) and primary lung fibroblasts: (i) viral recognition occurs chiefly via a Toll-like receptor (TLR)-independent nucleic acid-sensing mechanism recognizing the viral dsDNA genome, (ii) the intact viral particle and capsid proteins are required for efficient intracellular delivery of the viral genome, (iii) delivery of the viral genome triggers interferon regulatory factor 3 (IRF3) phosphorylation, and (iv) IRF3 activation is the required dominant antiviral signaling pathway used by APC, whereas the “primary” involvement of NF-κB, mitogen-activated protein kinase, or Akt pathways is less prominent. In this study we provide the first direct evidence that infection by a dsDNA virus stimulates an IRF3-mediated interferon and proinflammatory response through a TLR-independent DNA-sensing mechanism.
- Published
- 2007
47. Erratum: Corrigendum: A diverse range of gene products are effectors of the type I interferon antiviral response
- Author
-
John W. Schoggins, Mary Murphy, Paul D. Bieniasz, Sam J. Wilson, Christopher T. Jones, Charles M. Rice, and Maryline Panis
- Subjects
Multidisciplinary ,Effector ,West Nile virus ,Interferon ,Venezuelan equine encephalitis virus ,medicine ,Biology ,medicine.disease_cause ,Gene ,Virology ,Virus ,medicine.drug - Abstract
Nature 472, 481–485 (2011); doi:10.1038/nature09907 We have recently discovered that the WNV-GFP stock used in the data set (Fig. 2 and Supplementary Table 8 of the original Letter) for West Nile virus (WNV) in this Letter was actually Venezuelan equine encephalitis virus (VEEV-GFP). The error has been tracked to a technical mistake made during the virus production process.
- Published
- 2015
48. I04 Identification and characterization of antiviral interferon effectors across the viral phylogeny
- Author
-
John W. Schoggins
- Subjects
viruses ,Hepatitis C virus ,Immunology ,virus diseases ,RNA ,DNA virus ,Hematology ,Biology ,Dengue virus ,medicine.disease_cause ,Biochemistry ,Virology ,Viral life cycle ,Interferon ,Viral evolution ,Influenza A virus ,medicine ,Immunology and Allergy ,Molecular Biology ,medicine.drug - Abstract
The type I interferon (IFN) response protects cells from invading viral pathogens by triggering the transcription of interferon-stimulated genes (ISGs). Although hundreds of ISGs have been identified, relatively few have been characterized with respect to antiviral activity. For most, little is known about their antiviral potential, their target specificity, and their mechanisms of action. We have established an overexpression screening platform to evaluate the antiviral potential of more than 350 ISGs against a diverse panel of animal viruses spanning the viral phylogeny. To date, we have screened 13 plus-strand RNA viruses, 7 negative-strand RNA viruses, and one DNA virus. Hierarchical clustering of top ISG hits indicates that viruses segregate based on their genomic content, with plus-stranded RNA viruses exhibiting the highest sensitivity to the greatest number of ISGs. We have also probed the mechanisms of action of select antiviral ISGs against hepatitis C virus, dengue virus, and influenza A virus. Many of these ISGs feed back into antiviral programs, while others appear to have more targeted effector functions. Using a variety of experimental techniques, we show that inhibition of early viral life cycle events is a common mechanism targeted by numerous IFN effectors. Gaining additional insight into the cellular and biochemical mechanisms if ISG-mediated inhibition may provide a platform for the development of novel antiviral therapies.
- Published
- 2012
49. PS1-122 Unraveling the complexity of the type I interferon antiviral response
- Author
-
Mary Murphy, John W. Schoggins, Charles M. Rice, Christopher T. Jones, Sam J. Wilson, Paul D. Bieniasz, and Maryline Panis
- Subjects
Interferon ,Immunology ,medicine ,Immunology and Allergy ,Hematology ,Biology ,Molecular Biology ,Biochemistry ,Virology ,medicine.drug - Published
- 2011
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