PLoS Articles
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UNC-authored articles published by the Public Library of Science (PLoS)
Works (2112)
2091. MyD88 Is Required for Protection from Lethal Infection with a Mouse-Adapted SARS-CoV
- Title Tesim:
- MyD88 Is Required for Protection from Lethal Infection with a Mouse-Adapted SARS-CoV
- Creator:
- Baric, Ralph S., Sheahan, Timothy, Heise, Mark T., Akira, Shizou, Uematsu, Satoshi, Morrison, Thomas E., and Funkhouser, William
- Date of publication:
- 2008
- Abstract Tesim:
- A novel human coronavirus, SARS-CoV, emerged suddenly in 2003, causing approximately 8000 human cases and more than 700 deaths worldwide. Since most animal models fail to faithfully recapitulate the clinical course of SARS-CoV in humans, the virus and host factors that mediate disease pathogenesis remain unclear. Recently, our laboratory and others developed a recombinant mouse-adapted SARS-CoV (rMA15) that was lethal in BALB/c mice. In contrast, intranasal infection of young 10-week-old C57BL/6 mice with rMA15 results in a nonlethal infection characterized by high titer replication within the lungs, lung inflammation, destruction of lung tissue, and loss of body weight, thus providing a useful model to identify host mediators of protection. Here, we report that mice deficient in MyD88 (MyD88−/−), an adapter protein that mediates Toll-like receptor (TLR), IL-1R, and IL-18R signaling, are far more susceptible to rMA15 infection. The genetic absence of MyD88 resulted in enhanced pulmonary pathology and greater than 90% mortality by day 6 post-infection. MyD88−/− mice had significantly higher viral loads in lung tissue throughout the course of infection. Despite increased viral loads, the expression of multiple proinflammatory cytokines and chemokines within lung tissue and recruitment of inflammatory monocytes/macrophages to the lung was severely impaired in MyD88−/− mice compared to wild-type mice. Furthermore, mice deficient in chemokine receptors that contribute to monocyte recruitment to the lung were more susceptible to rMA15-induced disease and exhibited severe lung pathology similar to that seen in MyD88−/−mice. These data suggest that MyD88-mediated innate immune signaling and inflammatory cell recruitment to the lung are required for protection from lethal rMA15 infection.
- Resource type:
- Article
- Admin Note:
- deposited as part of the 1Science batch
- Affiliation Label Tesim:
- Department of Microbiology and Immunology, Department of Genetics, and Department of Pathology and Laboratory Medicine
- Type:
- http://purl.org/dc/dcmitype/Text
- DOI:
- https://doi.org/10.17615/9e08-dg24
- Identifier:
- https://dx.doi.org/10.1371/journal.ppat.1000240
- ISSN:
- 1553-7366
- Journal Issue:
- 12
- Journal Title:
- PLoS Pathogens
- Journal Volume:
- 4
- Language Label:
- English
- ORCID:
- Other Affiliation:
- and Osaka University
- Person:
- Baric, Ralph S., Sheahan, Timothy, Heise, Mark T., Akira, Shizou, Uematsu, Satoshi, Morrison, Thomas E., and Funkhouser, William
- Rights Statement Label:
- In Copyright
- Source:
- 8p58pj753
2092. A Network Integration Approach to Predict Conserved Regulators Related to Pathogenicity of Influenza and SARS-CoV Respiratory Viruses
- Title Tesim:
- A Network Integration Approach to Predict Conserved Regulators Related to Pathogenicity of Influenza and SARS-CoV Respiratory Viruses
- Creator:
- Benecke, Arndt G., Tchitchek, Nicolas, Mitchell, Hugh D., Luna, Maria L., Smith, Richard D., McDermott, Jason E., Ellis, Amy L., Neumann, Gabriele, Sims, Amy C., Katze, Michael G., Kawaoka, Yoshihiro, Tilton, Susan C., Metz, Thomas O., Waters, Katrina M., Eisfeld, Amie J., Matzke, Melissa M., Josset, Laurence, Li, Chengjun, Chang, Jean H., Baric, Ralph S., Schepmoes, Athena A., Shukla, Anil K., Heegel, Robert A., and Webb-Robertson, Bobbi-Jo M.
- Date of publication:
- 2013
- Abstract Tesim:
- Respiratory infections stemming from influenza viruses and the Severe Acute Respiratory Syndrome corona virus (SARS-CoV) represent a serious public health threat as emerging pandemics. Despite efforts to identify the critical interactions of these viruses with host machinery, the key regulatory events that lead to disease pathology remain poorly targeted with therapeutics. Here we implement an integrated network interrogation approach, in which proteome and transcriptome datasets from infection of both viruses in human lung epithelial cells are utilized to predict regulatory genes involved in the host response. We take advantage of a novel "crowd-based" approach to identify and combine ranking metrics that isolate genes/proteins likely related to the pathogenicity of SARS-CoV and influenza virus. Subsequently, a multivariate regression model is used to compare predicted lung epithelial regulatory influences with data derived from other respiratory virus infection models. We predicted a small set of regulatory factors with conserved behavior for consideration as important components of viral pathogenesis that might also serve as therapeutic targets for intervention. Our results demonstrate the utility of integrating diverse 'omic datasets to predict and prioritize regulatory features conserved across multiple pathogen infection models.
- Resource type:
- Article
- Admin Note:
- deposited as part of the 1Science batch
- Affiliation Label Tesim:
- Department of Epidemiology
- Type:
- http://purl.org/dc/dcmitype/Text
- DOI:
- https://doi.org/10.17615/mbhe-hg94
- Identifier:
- https://dx.doi.org/10.1371/journal.pone.0069374
- ISSN:
- 1932-6203
- Journal Issue:
- 7
- Journal Title:
- PloS One
- Journal Volume:
- 8
- Language Label:
- English
- ORCID:
- Other Affiliation:
- University of Washington, Pacific Northwest National Laboratory, University of Wisconsin-Madison, and
- Person:
- Benecke, Arndt G., Tchitchek, Nicolas, Mitchell, Hugh D., Luna, Maria L., Smith, Richard D., McDermott, Jason E., Ellis, Amy L., Neumann, Gabriele, Sims, Amy C., Katze, Michael G., Kawaoka, Yoshihiro, Tilton, Susan C., Metz, Thomas O., Waters, Katrina M., Eisfeld, Amie J., Matzke, Melissa M., Josset, Laurence, Li, Chengjun, Chang, Jean H., Baric, Ralph S., Schepmoes, Athena A., Shukla, Anil K., Heegel, Robert A., and Webb-Robertson, Bobbi-Jo M.
- Rights Statement Label:
- In Copyright
- Source:
- kd17cz846
2093. Yeast Based Small Molecule Screen for Inhibitors of SARS-CoV
- Title Tesim:
- Yeast Based Small Molecule Screen for Inhibitors of SARS-CoV
- Creator:
- Jones, Grant, Matthews, Krystal, Pickles, Raymond, Basu, Dipanwita, Taylor, Justin, Frieman, Matthew, Baric, Ralph S., and Engel, Daniel A.
- Date of publication:
- 2011
- Abstract Tesim:
- Severe acute respiratory coronavirus (SARS-CoV) emerged in 2002, resulting in roughly 8000 cases worldwide and 10% mortality. The animal reservoirs for SARS-CoV precursors still exist and the likelihood of future outbreaks in the human population is high. The SARS-CoV papain-like protease (PLP) is an attractive target for pharmaceutical development because it is essential for virus replication and is conserved among human coronaviruses. A yeast-based assay was established for PLP activity that relies on the ability of PLP to induce a pronounced slow-growth phenotype when expressed in S. cerevisiae. Induction of the slow-growth phenotype was shown to take place over a 60-hour time course, providing the basis for conducting a screen for small molecules that restore growth by inhibiting the function of PLP. Five chemical suppressors of the slow-growth phenotype were identified from the 2000 member NIH Diversity Set library. One of these, NSC158362, potently inhibited SARS-CoV replication in cell culture without toxic effects on cells, and it specifically inhibited SARS-CoV replication but not influenza virus replication. The effect of NSC158362 on PLP protease, deubiquitinase and anti-interferon activities was investigated but the compound did not alter these activities. Another suppressor, NSC158011, demonstrated the ability to inhibit PLP protease activity in a cell-based assay. The identification of these inhibitors demonstrated a strong functional connection between the PLP-based yeast assay, the inhibitory compounds, and SARS-CoV biology. Furthermore the data with NSC158362 suggest a novel mechanism for inhibition of SARS-CoV replication that may involve an unknown activity of PLP, or alternatively a direct effect on a cellular target that modifies or bypasses PLP function in yeast and mammalian cells.
- Resource type:
- Article
- Admin Note:
- deposited as part of 1Science batch
- Affiliation Label Tesim:
- Gene Therapy Center and Gillings School of Global Public Health
- Type:
- http://purl.org/dc/dcmitype/Text
- DOI:
- https://doi.org/10.17615/sds4-5721
- Identifier:
- https://dx.doi.org/10.1371/journal.pone.0028479
- ISSN:
- 1932-6203
- Journal Issue:
- 12
- Journal Title:
- PloS One
- Journal Volume:
- 6
- Language Label:
- English
- ORCID:
- Other Affiliation:
- University of Maryland, , and University of Virginia School of Medicine
- Person:
- Jones, Grant, Matthews, Krystal, Pickles, Raymond, Basu, Dipanwita, Taylor, Justin, Frieman, Matthew, Baric, Ralph S., and Engel, Daniel A.
- Rights Statement Label:
- In Copyright
- Source:
- v979v777t
2094. Comparative Pathogenesis of Three Human and Zoonotic SARS-CoV Strains in Cynomolgus Macaques
- Title Tesim:
- Comparative Pathogenesis of Three Human and Zoonotic SARS-CoV Strains in Cynomolgus Macaques
- Creator:
- Porcella, Stephen F., Feldmann, Friederike, Feldmann, Heinz, Kercher, Lisa, Baric, Ralph S., Rosenke, Rebecca, Viraneva, Kimmo, Brining, Douglas, Mattoon, John, Rockx, Barry, LaCasse, Rachel, Parnell, Michael, Sturdevant, Daniel E., and Long, Dan
- Date of publication:
- 2011
- Abstract Tesim:
- The severe acute respiratory syndrome (SARS) epidemic was characterized by increased pathogenicity in the elderly due to an early exacerbated innate host response. SARS-CoV is a zoonotic pathogen that entered the human population through an intermediate host like the palm civet. To prevent future introductions of zoonotic SARS-CoV strains and subsequent transmission into the human population, heterologous disease models are needed to test the efficacy of vaccines and therapeutics against both late human and zoonotic isolates. Here we show that both human and zoonotic SARS-CoV strains can infect cynomolgus macaques and resulted in radiological as well as histopathological changes similar to those seen in mild human cases. Viral replication was higher in animals infected with a late human phase isolate compared to a zoonotic isolate. While there were significant differences in the number of host genes differentially regulated during the host responses between the three SARS-CoV strains, the top pathways and functions were similar and only apparent early during infection with the majority of genes associated with interferon signaling pathways. This study characterizes critical disease models in the evaluation and licensure of therapeutic strategies against SARS-CoV for human use.
- Resource type:
- Article
- Admin Note:
- deposited as part of the 1Science batch
- Affiliation Label Tesim:
- Department of Epidemiology
- Type:
- http://purl.org/dc/dcmitype/Text
- DOI:
- https://doi.org/10.17615/hz4z-jn29
- Identifier:
- https://dx.doi.org/10.1371/journal.pone.0018558
- ISSN:
- 1932-6203
- Journal Issue:
- 4
- Journal Title:
- PloS One
- Journal Volume:
- 6
- Language Label:
- English
- ORCID:
- Other Affiliation:
- National Institutes of Health, , and Washington State University
- Person:
- Porcella, Stephen F., Feldmann, Friederike, Feldmann, Heinz, Kercher, Lisa, Baric, Ralph S., Rosenke, Rebecca, Viraneva, Kimmo, Brining, Douglas, Mattoon, John, Rockx, Barry, LaCasse, Rachel, Parnell, Michael, Sturdevant, Daniel E., and Long, Dan
- Rights Statement Label:
- In Copyright
- Source:
- t722hf60h
2095. Vaccine Efficacy in Senescent Mice Challenged with Recombinant SARS-CoV Bearing Epidemic and Zoonotic Spike Variants
- Title Tesim:
- Vaccine Efficacy in Senescent Mice Challenged with Recombinant SARS-CoV Bearing Epidemic and Zoonotic Spike Variants
- Creator:
- Whitmore, Alan, West, Ande, Baric, Ralph S., Harkema, Jack, Johnston, Robert, Sims, Amy, Pickles, Raymond, Suthar, Mehul, Deming, Damon, Yount, Boyd, Davis, Nancy, Curtis, Kristopher, Heise, Mark, Donaldson, Eric, and Sheahan, Timothy
- Date of publication:
- 2006
- Abstract Tesim:
- Background: In 2003, severe acute respiratory syndrome coronavirus (SARS-CoV) was identified as the etiological agent of severe acute respiratory syndrome, a disease characterized by severe pneumonia that sometimes results in death. SARS-CoV is a zoonotic virus that crossed the species barrier, most likely originating from bats or from other species including civets, raccoon dogs, domestic cats, swine, and rodents. A SARS-CoV vaccine should confer long-term protection, especially in vulnerable senescent populations, against both the 2003 epidemic strains and zoonotic strains that may yet emerge from animal reservoirs. We report the comprehensive investigation of SARS vaccine efficacy in young and senescent mice following homologous and heterologous challenge. Methods and Findings: Using Venezuelan equine encephalitis virus replicon particles (VRP) expressing the 2003 epidemic Urbani SARS-CoV strain spike (S) glycoprotein (VRP-S) or the nucleocapsid (N) protein from the same strain (VRP-N), we demonstrate that VRP-S, but not VRP-N vaccines provide complete short- and long-term protection against homologous strain challenge in young and senescent mice. To test VRP vaccine efficacy against a heterologous SARS-CoV, we used phylogenetic analyses, synthetic biology, and reverse genetics to construct a chimeric virus (icGDO3-S) encoding a synthetic S glycoprotein gene of the most genetically divergent human strain, GDO3, which clusters among the zoonotic SARS-CoV. icGD03-S replicated efficiently in human airway epithelial cells and in the lungs of young and senescent mice, and was highly resistant to neutralization with antisera directed against the Urbani strain. Although VRP-S vaccines provided complete short-term protection against heterologous icGD03-S challenge in young mice, only limited protection was seen in vaccinated senescent animals. VRP-N vaccines not only failed to protect from homologous or heterologous challenge, but resulted in enhanced immunopathology with eosinophilic infiltrates within the lungs of SARS-CoV–challenged mice. VRP-N–induced pathology presented at day 4, peaked around day 7, and persisted through day 14, and was likely mediated by cellular immune responses. Conclusions: This study identifies gaps and challenges in vaccine design for controlling future SARS-CoV zoonosis, especially in vulnerable elderly populations. The availability of a SARS-CoV virus bearing heterologous S glycoproteins provides a robust challenge inoculum for evaluating vaccine efficacy against zoonotic strains, the most likely source of future outbreaks.
- Resource type:
- Article
- Admin Note:
- deposited as part of the 1Science batch
- Affiliation Label Tesim:
- Carolina Vaccine Institute, Department of Microbiology and Immunology, and Department of Epidemiology
- Type:
- http://purl.org/dc/dcmitype/Text
- DOI:
- https://doi.org/10.17615/72cq-r833
- Identifier:
- https://doi.org/10.1371/journal.pmed.0030525
- ISSN:
- 1549-1277
- Journal Issue:
- 12
- Journal Title:
- PLoS Medicine
- Journal Volume:
- 3
- Language Label:
- English
- ORCID:
- Other Affiliation:
- , Michigan State University, and United States Army Medical Research Institute of Infectious Diseases
- Person:
- Whitmore, Alan, West, Ande, Baric, Ralph S., Harkema, Jack, Johnston, Robert, Sims, Amy, Pickles, Raymond, Suthar, Mehul, Deming, Damon, Yount, Boyd, Davis, Nancy, Curtis, Kristopher, Heise, Mark, Donaldson, Eric, and Sheahan, Timothy
- Rights Statement Label:
- In Copyright
- Source:
- sj139694g
2096. Genome Wide Identification of SARS-CoV Susceptibility Loci Using the Collaborative Cross
- Title Tesim:
- Genome Wide Identification of SARS-CoV Susceptibility Loci Using the Collaborative Cross
- Creator:
- Bell, Timothy A., Katze, Michael G., Menachery, Vineet D., Whitmore, Alan C., Threadgill, David W., Buus, Ryan J., Ferris, Martin T., Gralinski, Lisa E., Deming, Damon, Valdar, William, Baric, Ralph S., Green, Richard, McMillan, Leonard, Frieman, Matthew B., de Villena, Fernando Pardo-Manuel, Miller, Darla R., Aylor, David L., Heise, Mark T., and Churchill, Gary A.
- Date of publication:
- 2015
- Abstract Tesim:
- New systems genetics approaches are needed to rapidly identify host genes and genetic networks that regulate complex disease outcomes. Using genetically diverse animals from incipient lines of the Collaborative Cross mouse panel, we demonstrate a greatly expanded range of phenotypes relative to classical mouse models of SARS-CoV infection including lung pathology, weight loss and viral titer. Genetic mapping revealed several loci contributing to differential disease responses, including an 8.5Mb locus associated with vascular cuffing on chromosome 3 that contained 23 genes and 13 noncoding RNAs. Integrating phenotypic and genetic data narrowed this region to a single gene, Trim55, an E3 ubiquitin ligase with a role in muscle fiber maintenance. Lung pathology and transcriptomic data from mice genetically deficient in Trim55 were used to validate its role in SARS-CoV-induced vascular cuffing and inflammation. These data establish the Collaborative Cross platform as a powerful genetic resource for uncovering genetic contributions of complex traits in microbial disease severity, inflammation and virus replication in models of outbred populations.
- Resource type:
- Article
- Admin Note:
- deposited as part of the 1Science batch
- Affiliation Label Tesim:
- Department of Genetics, Department of Epidemiology, and Department of Computer Science
- Type:
- http://purl.org/dc/dcmitype/Text
- DOI:
- https://doi.org/10.17615/z06v-7412
- Identifier:
- https://dx.doi.org/10.1371/journal.pgen.1005504
- ISSN:
- 1553-7390
- Journal Issue:
- 10
- Journal Title:
- PLoS Genetics
- Journal Volume:
- 11
- Language Label:
- English
- ORCID:
- Other Affiliation:
- , University of Washington, Texas A&M University, and The Jackson Laboratory
- Person:
- Bell, Timothy A., Katze, Michael G., Menachery, Vineet D., Whitmore, Alan C., Threadgill, David W., Buus, Ryan J., Ferris, Martin T., Gralinski, Lisa E., Deming, Damon, Valdar, William, Baric, Ralph S., Green, Richard, McMillan, Leonard, Frieman, Matthew B., de Villena, Fernando Pardo-Manuel, Miller, Darla R., Aylor, David L., Heise, Mark T., and Churchill, Gary A.
- Rights Statement Label:
- In Copyright
- Source:
- pn89dc09w
2097. Correction: SARS Coronavirus 3b Accessory Protein Modulates Transcriptional Activity of RUNX1b
- Title Tesim:
- Correction: SARS Coronavirus 3b Accessory Protein Modulates Transcriptional Activity of RUNX1b
- Creator:
- Ralph Baric, Bhavna Varshney, Sunil K. Lal, Sudhakar Agnihotram, and Yee-Joo Tan
- Date of publication:
- 2012
- Abstract Tesim:
- BackgroundThe causative agent of severe acute respiratory syndrome, SARS coronavirus (SARS-CoV) genome encodes several unique group specific accessory proteins with unknown functions. Among them, accessory protein 3b (also known as ORF4) was lately identified as one of the viral interferon antagonist. Recently our lab uncovered a new role for 3b in upregulation of AP-1 transcriptional activity and its downstream genes. Thus, we believe that 3b might play an important role in SARS-CoV pathogenesis and therefore is of considerable interest. The current study aims at identifying novel host cellular interactors of the 3b protein.Methodology/Principal FindingsIn this study, using yeast two-hybrid and co-immunoprecipitation techniques, we have identified a host transcription factor RUNX1b (Runt related transcription factor, isoform b) as a novel interacting partner for SARS-CoV 3b protein. Chromatin immunoprecipitaion (ChIP) and reporter gene assays in 3b expressing jurkat cells showed recruitment of 3b on the RUNX1 binding element that led to an increase in RUNX1b transactivation potential on the IL2 promoter. Kinase assay and pharmacological inhibitor treatment implied that 3b also affect RUNX1b transcriptional activity by regulating its ERK dependent phosphorylation levels. Additionally, mRNA levels of MIP-1α, a RUNX1b target gene upregulated in SARS-CoV infected monocyte-derived dendritic cells, were found to be elevated in 3b expressing U937 monocyte cells.Conclusions/SignificanceThese results unveil a novel interaction of SARS-CoV 3b with the host factor, RUNX1b, and speculate its physiological relevance in upregulating cytokines and chemokine levels in state of SARS virus infection.
- Resource type:
- Article
- Type:
- http://purl.org/dc/dcmitype/Text
- DOI:
- https://doi.org/10.17615/65r0-7a56
- ISSN:
- 1932-6203
- Journal Issue:
- 1
- Journal Title:
- PLoS ONE
- Journal Volume:
- 7
- Keyword:
- Enzyme Activation, Phosphorylation, RNA, Messenger, Binding Sites, Cell Nucleus, Chemokine CCL3, Humans, Core Binding Factor Alpha 2 Subunit, Promoter Regions, Genetic, Animals, Protein Binding, Interleukin-2, Extracellular Signal-Regulated MAP Kinases, Mice, and Cell Line
- Language Label:
- English
- ORCID:
- Other Affiliation:
- Department of Microbiology and Immunology, University of North Carolina, Chapel Hill, North Carolina, United States of America, Virology Group, International Centre for Genetic Engineering and Biotechnology, New Delhi, India, and Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore
- Page End:
- 9
- Page Start:
- 1
- Person:
- Ralph Baric, Bhavna Varshney, Sunil K. Lal, Sudhakar Agnihotram, and Yee-Joo Tan
- Rights Statement Label:
- In Copyright
- Source:
- 3b591d98k
2098. A Mouse-Adapted SARS-Coronavirus Causes Disease and Mortality in BALB/c Mice
- Title Tesim:
- A Mouse-Adapted SARS-Coronavirus Causes Disease and Mortality in BALB/c Mice
- Creator:
- Yount, Boyd, Heise, Mark, Sheahan, Tim, Baric, Ralph, Deming, Damon, Roberts, Anjeanette, Subbararo, Kanta, Cheng, Aaron, Genrich, Gillian L., Zaki, Sherif R., Herman, Brian D., Paddock, Christopher, and Vogel, Leatrice
- Date of publication:
- 2007
- Abstract Tesim:
- No single animal model for severe acute respiratory syndrome (SARS) reproduces all aspects of the human disease. Young inbred mice support SARS-coronavirus (SARS-CoV) replication in the respiratory tract and are available in sufficient numbers for statistical evaluation. They are relatively inexpensive and easily accessible, but their use in SARS research is limited because they do not develop illness following infection. Older (12- to 14-mo-old) BALB/c mice develop clinical illness and pneumonitis, but they can be hard to procure, and immune senescence complicates pathogenesis studies. We adapted the SARS-CoV (Urbani strain) by serial passage in the respiratory tract of young BALB/c mice. Fifteen passages resulted in a virus (MA15) that is lethal for mice following intranasal inoculation. Lethality is preceded by rapid and high titer viral replication in lungs, viremia, and dissemination of virus to extrapulmonary sites accompanied by lymphopenia, neutrophilia, and pathological changes in the lungs. Abundant viral antigen is extensively distributed in bronchial epithelial cells and alveolar pneumocytes, and necrotic cellular debris is present in airways and alveoli, with only mild and focal pneumonitis. These observations suggest that mice infected with MA15 die from an overwhelming viral infection with extensive, virally mediated destruction of pneumocytes and ciliated epithelial cells. The MA15 virus has six coding mutations associated with adaptation and increased virulence; when introduced into a recombinant SARS-CoV, these mutations result in a highly virulent and lethal virus (rMA15), duplicating the phenotype of the biologically derived MA15 virus. Intranasal inoculation with MA15 reproduces many aspects of disease seen in severe human cases of SARS. The availability of the MA15 virus will enhance the use of the mouse model for SARS because infection with MA15 causes morbidity, mortality, and pulmonary pathology. This virus will be of value as a stringent challenge in evaluation of the efficacy of vaccines and antivirals.
- Resource type:
- Article
- Admin Note:
- deposited as part of 1Science uploads
- Affiliation Label Tesim:
- Department of Microbiology and Immunology
- Type:
- http://purl.org/dc/dcmitype/Text
- DOI:
- https://doi.org/10.17615/5ehf-dw39
- Identifier:
- https://dx.doi.org/10.1371/journal.ppat.0030005
- ISSN:
- 1553-7366
- Journal Issue:
- 1
- Journal Title:
- PLoS Pathogens
- Journal Volume:
- 3
- Keyword:
- SARS Virus, RNA Virus Infections, Humans, Disease Models, Animal, Mice, Inbred BALB C, Molecular Sequence Data, Animals, Mice, and Severe Acute Respiratory Syndrome
- Language Label:
- English
- ORCID:
- Other Affiliation:
- , National Institutes of Health, and Centers for Disease Control and Prevention
- Page Start:
- e5
- Person:
- Yount, Boyd, Heise, Mark, Sheahan, Tim, Baric, Ralph, Deming, Damon, Roberts, Anjeanette, Subbararo, Kanta, Cheng, Aaron, Genrich, Gillian L., Zaki, Sherif R., Herman, Brian D., Paddock, Christopher, and Vogel, Leatrice
- Rights Statement Label:
- In Copyright
- Source:
- 3n204358b
2099. Analysis of Intraviral Protein-Protein Interactions of the SARS Coronavirus ORFeome
- Title Tesim:
- Analysis of Intraviral Protein-Protein Interactions of the SARS Coronavirus ORFeome
- Creator:
- Teepe, Carola, Simpson, Jeremy, Friedel, Caroline C., Pepperkok, Rainer, Roberts, Rhonda, Haas, Juergen, von Brunn, Albrecht, Baric, Ralph, and Zimmer, Ralf
- Date of publication:
- 2007
- Abstract Tesim:
- The severe acute respiratory syndrome coronavirus (SARS-CoV) genome is predicted to encode 14 functional open reading frames, leading to the expression of up to 30 structural and non-structural protein products. The functions of a large number of viral ORFs are poorly understood or unknown. In order to gain more insight into functions and modes of action and interaction of the different proteins, we cloned the viral ORFeome and performed a genome-wide analysis for intraviral protein interactions and for intracellular localization. 900 pairwise interactions were tested by yeast-two-hybrid matrix analysis, and more than 65 positive non-redundant interactions, including six self interactions, were identified. About 38% of interactions were subsequently confirmed by CoIP in mammalian cells. Nsp2, nsp8 and ORF9b showed a wide range of interactions with other viral proteins. Nsp8 interacts with replicase proteins nsp2, nsp5, nsp6, nsp7, nsp8, nsp9, nsp12, nsp13 and nsp14, indicating a crucial role as a major player within the replication complex machinery. It was shown by others that nsp8 is essential for viral replication in vitro, whereas nsp2 is not. We show that also accessory protein ORF9b does not play a pivotal role for viral replication, as it can be deleted from the virus displaying normal plaque sizes and growth characteristics in Vero cells. However, it can be expected to be important for the virus-host interplay and for pathogenicity, due to its large number of interactions, by enhancing the global stability of the SARS proteome network, or play some unrealized role in regulating protein-protein interactions. The interactions identified provide valuable material for future studies.
- Resource type:
- Article
- Admin Note:
- deposited as part of the 1Science upload
- Affiliation Label Tesim:
- University of North Carolina at Chapel Hill
- Type:
- http://purl.org/dc/dcmitype/Text
- DOI:
- https://doi.org/10.17615/bzbk-w739
- Identifier:
- https://dx.doi.org/10.1371/journal.pone.0000459
- ISSN:
- 1932-6203
- Journal Issue:
- 5
- Journal Title:
- PloS One
- Journal Volume:
- 2
- Keyword:
- Viral Proteins, Molecular Sequence Data, Fluorescent Antibody Technique, Genome, Amino Acid Sequence, Open Reading Frames, and SARS Virus
- Language Label:
- English
- ORCID:
- Other Affiliation:
- Ludwig-Maximilians-Universitaet, European Molecular Biology Laboratory, and
- Page Start:
- e459
- Person:
- Teepe, Carola, Simpson, Jeremy, Friedel, Caroline C., Pepperkok, Rainer, Roberts, Rhonda, Haas, Juergen, von Brunn, Albrecht, Baric, Ralph, and Zimmer, Ralf
- Rights Statement Label:
- In Copyright
- Source:
- xk81jq99t
2100. Zn2+ Inhibits Coronavirus and Arterivirus RNA Polymerase Activity In Vitro and Zinc Ionophores Block the Replication of These Viruses in Cell Culture
- Title Tesim:
- Zn2+ Inhibits Coronavirus and Arterivirus RNA Polymerase Activity In Vitro and Zinc Ionophores Block the Replication of These Viruses in Cell Culture
- Creator:
- van den Worm, Sjoerd H.E., Sims, Amy C., Baric, Ralph S., te Velthuis, Aartjan J.W., Snijder, Eric J., and van Hemert, Martijn J.
- Date of publication:
- 2010
- Abstract Tesim:
- Increasing the intracellular Zn2+ concentration with zinc-ionophores like pyrithione (PT) can efficiently impair the replication of a variety of RNA viruses, including poliovirus and influenza virus. For some viruses this effect has been attributed to interference with viral polyprotein processing. In this study we demonstrate that the combination of Zn2+ and PT at low concentrations (2 µM Zn2+ and 2 µM PT) inhibits the replication of SARS-coronavirus (SARS-CoV) and equine arteritis virus (EAV) in cell culture. The RNA synthesis of these two distantly related nidoviruses is catalyzed by an RNA-dependent RNA polymerase (RdRp), which is the core enzyme of their multiprotein replication and transcription complex (RTC). Using an activity assay for RTCs isolated from cells infected with SARS-CoV or EAV—thus eliminating the need for PT to transport Zn2+ across the plasma membrane—we show that Zn2+ efficiently inhibits the RNA-synthesizing activity of the RTCs of both viruses. Enzymatic studies using recombinant RdRps (SARS-CoV nsp12 and EAV nsp9) purified from E. coli subsequently revealed that Zn2+ directly inhibited the in vitro activity of both nidovirus polymerases. More specifically, Zn2+ was found to block the initiation step of EAV RNA synthesis, whereas in the case of the SARS-CoV RdRp elongation was inhibited and template binding reduced. By chelating Zn2+ with MgEDTA, the inhibitory effect of the divalent cation could be reversed, which provides a novel experimental tool for in vitro studies of the molecular details of nidovirus replication and transcription.
- Resource type:
- Article
- Admin Note:
- deposited as part of the 1Science upload
- Affiliation Label Tesim:
- Department of Epidemiology
- Type:
- http://purl.org/dc/dcmitype/Text
- DOI:
- https://doi.org/10.17615/rvb5-9y41
- Identifier:
- https://dx.doi.org/10.1371/journal.ppat.1001176
- ISSN:
- 1553-7366
- Journal Issue:
- 11
- Journal Title:
- PLoS Pathogens
- Journal Volume:
- 6
- Keyword:
- Ionophores, RNA, Messenger, Vero Cells, Zinc Compounds, Severe Acute Respiratory Syndrome, RNA Replicase, Escherichia coli, Virus Replication, Animals, Cercopithecus aethiops, Arterivirus Infections, Reverse Transcriptase Polymerase Chain Reaction, Electrophoretic Mobility Shift Assay, Blotting, Western, SARS Virus, RNA, Viral, and Arterivirus
- Language Label:
- English
- ORCID:
- Other Affiliation:
- Leiden University Medical Center and
- Page Start:
- e1001176
- Person:
- van den Worm, Sjoerd H.E., Sims, Amy C., Baric, Ralph S., te Velthuis, Aartjan J.W., Snijder, Eric J., and van Hemert, Martijn J.
- Rights Statement Label:
- In Copyright
- Source:
- zw12z9633
Collection Details
- Total items
-
2112
- Size
-
unknown
- Date created
-
February 2, 2022