Yeast Based Small Molecule Screen for Inhibitors of SARS-CoV
Identifieur interne : 002008 ( Main/Exploration ); précédent : 002007; suivant : 002009Yeast Based Small Molecule Screen for Inhibitors of SARS-CoV
Auteurs : Matthew Frieman [États-Unis] ; Dipanwita Basu [États-Unis] ; Krystal Matthews [États-Unis] ; Justin Taylor [États-Unis] ; Grant Jones [États-Unis] ; Raymond Pickles [États-Unis] ; Ralph Baric [États-Unis] ; Daniel A. Engel [États-Unis]Source :
- PLoS ONE [ 1932-6203 ] ; 2011.
Descripteurs français
- KwdFr :
- Animaux, Antiviraux (pharmacologie), Cellules HEK293, Cellules Vero, Cellules épithéliales (cytologie), Clonage moléculaire, Conception de médicament, Humains, Liaison aux protéines, Milieux de culture (métabolisme), Orthomyxoviridae (génétique), Phénotype, Protéines à fluorescence verte (métabolisme), Saccharomyces cerevisiae (métabolisme), Saccharomyces cerevisiae (virologie), Techniques in vitro, Technologie pharmaceutique (), Trachée (métabolisme), Virus du SRAS (métabolisme).
- MESH :
- cytologie : Cellules épithéliales.
- génétique : Orthomyxoviridae.
- métabolisme : Milieux de culture, Protéines à fluorescence verte, Saccharomyces cerevisiae, Trachée, Virus du SRAS.
- pharmacologie : Antiviraux.
- virologie : Saccharomyces cerevisiae.
- Animaux, Cellules HEK293, Cellules Vero, Clonage moléculaire, Conception de médicament, Humains, Liaison aux protéines, Phénotype, Techniques in vitro, Technologie pharmaceutique.
English descriptors
- KwdEn :
- Animals, Antiviral Agents (pharmacology), Chlorocebus aethiops, Cloning, Molecular, Culture Media (metabolism), Drug Design, Epithelial Cells (cytology), Green Fluorescent Proteins (metabolism), HEK293 Cells, Humans, In Vitro Techniques, Orthomyxoviridae (genetics), Phenotype, Protein Binding, SARS Virus (metabolism), Saccharomyces cerevisiae (metabolism), Saccharomyces cerevisiae (virology), Technology, Pharmaceutical (methods), Trachea (metabolism), Vero Cells.
- MESH :
- chemical , metabolism : Culture Media, Green Fluorescent Proteins.
- chemical , pharmacology : Antiviral Agents.
- cytology : Epithelial Cells.
- genetics : Orthomyxoviridae.
- metabolism : SARS Virus, Saccharomyces cerevisiae, Trachea.
- methods : Technology, Pharmaceutical.
- virology : Saccharomyces cerevisiae.
- Animals, Chlorocebus aethiops, Cloning, Molecular, Drug Design, HEK293 Cells, Humans, In Vitro Techniques, Phenotype, Protein Binding, Vero Cells.
Abstract
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
Url:
DOI: 10.1371/journal.pone.0028479
PubMed: 22164298
PubMed Central: 3229576
Affiliations:
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Le document en format XML
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<term>Chlorocebus aethiops</term>
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<term>Drug Design</term>
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<term>Green Fluorescent Proteins (metabolism)</term>
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<term>In Vitro Techniques</term>
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<term>SARS Virus (metabolism)</term>
<term>Saccharomyces cerevisiae (metabolism)</term>
<term>Saccharomyces cerevisiae (virology)</term>
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<term>Cellules Vero</term>
<term>Cellules épithéliales (cytologie)</term>
<term>Clonage moléculaire</term>
<term>Conception de médicament</term>
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<term>Liaison aux protéines</term>
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<front><div type="abstract" xml:lang="en"><p>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 <italic>S. cerevisiae</italic>
. 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.</p>
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</back>
</TEI>
<affiliations><list><country><li>États-Unis</li>
</country>
<region><li>Caroline du Nord</li>
<li>Maryland</li>
<li>Virginie</li>
</region>
<settlement><li>College Park (Maryland)</li>
</settlement>
<orgName><li>Université du Maryland</li>
</orgName>
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<name sortKey="Taylor, Justin" sort="Taylor, Justin" uniqKey="Taylor J" first="Justin" last="Taylor">Justin Taylor</name>
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