Yeast based small molecule screen for inhibitors of SARS-CoV.
Identifieur interne : 001433 ( PubMed/Corpus ); précédent : 001432; suivant : 001434Yeast based small molecule screen for inhibitors of SARS-CoV.
Auteurs : Matthew Frieman ; Dipanwita Basu ; Krystal Matthews ; Justin Taylor ; Grant Jones ; Raymond Pickles ; Ralph Baric ; Daniel A. EngelSource :
- PloS one [ 1932-6203 ] ; 2011.
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 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.
DOI: 10.1371/journal.pone.0028479
PubMed: 22164298
Links to Exploration step
pubmed:22164298Le document en format XML
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Engel</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2011">2011</date><idno type="RBID">pubmed:22164298</idno><idno type="pmid">22164298</idno><idno type="doi">10.1371/journal.pone.0028479</idno><idno type="wicri:Area/PubMed/Corpus">001433</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001433</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Yeast based small molecule screen for inhibitors of SARS-CoV.</title><author><name sortKey="Frieman, Matthew" sort="Frieman, Matthew" uniqKey="Frieman M" first="Matthew" last="Frieman">Matthew Frieman</name><affiliation><nlm:affiliation>Department of Microbiology and Immunology, University of Maryland, Baltimore, Maryland, United States of America.</nlm:affiliation></affiliation></author><author><name sortKey="Basu, Dipanwita" sort="Basu, Dipanwita" uniqKey="Basu D" first="Dipanwita" last="Basu">Dipanwita Basu</name></author><author><name sortKey="Matthews, Krystal" sort="Matthews, Krystal" uniqKey="Matthews K" first="Krystal" last="Matthews">Krystal Matthews</name></author><author><name sortKey="Taylor, Justin" sort="Taylor, Justin" uniqKey="Taylor J" first="Justin" last="Taylor">Justin Taylor</name></author><author><name sortKey="Jones, Grant" sort="Jones, Grant" uniqKey="Jones G" first="Grant" last="Jones">Grant Jones</name></author><author><name sortKey="Pickles, Raymond" sort="Pickles, Raymond" uniqKey="Pickles R" first="Raymond" last="Pickles">Raymond Pickles</name></author><author><name sortKey="Baric, Ralph" sort="Baric, Ralph" uniqKey="Baric R" first="Ralph" last="Baric">Ralph Baric</name></author><author><name sortKey="Engel, Daniel A" sort="Engel, Daniel A" uniqKey="Engel D" first="Daniel A" last="Engel">Daniel A. Engel</name></author></analytic><series><title level="j">PloS one</title><idno type="eISSN">1932-6203</idno><imprint><date when="2011" type="published">2011</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Antiviral Agents (pharmacology)</term><term>Chlorocebus aethiops</term><term>Cloning, Molecular</term><term>Culture Media (metabolism)</term><term>Drug Design</term><term>Epithelial Cells (cytology)</term><term>Green Fluorescent Proteins (metabolism)</term><term>HEK293 Cells</term><term>Humans</term><term>In Vitro Techniques</term><term>Orthomyxoviridae (genetics)</term><term>Phenotype</term><term>Protein Binding</term><term>SARS Virus (metabolism)</term><term>Saccharomyces cerevisiae (metabolism)</term><term>Saccharomyces cerevisiae (virology)</term><term>Technology, Pharmaceutical (methods)</term><term>Trachea (metabolism)</term><term>Vero Cells</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Culture Media</term><term>Green Fluorescent Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Antiviral Agents</term></keywords><keywords scheme="MESH" qualifier="cytology" xml:lang="en"><term>Epithelial Cells</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Orthomyxoviridae</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>SARS Virus</term><term>Saccharomyces cerevisiae</term><term>Trachea</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Technology, Pharmaceutical</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Saccharomyces cerevisiae</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Chlorocebus aethiops</term><term>Cloning, Molecular</term><term>Drug Design</term><term>HEK293 Cells</term><term>Humans</term><term>In Vitro Techniques</term><term>Phenotype</term><term>Protein Binding</term><term>Vero Cells</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">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.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">22164298</PMID><DateCompleted><Year>2012</Year><Month>07</Month><Day>20</Day></DateCompleted><DateRevised><Year>2019</Year><Month>12</Month><Day>10</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1932-6203</ISSN><JournalIssue CitedMedium="Internet"><Volume>6</Volume><Issue>12</Issue><PubDate><Year>2011</Year></PubDate></JournalIssue><Title>PloS one</Title><ISOAbbreviation>PLoS ONE</ISOAbbreviation></Journal><ArticleTitle>Yeast based small molecule screen for inhibitors of SARS-CoV.</ArticleTitle><Pagination><MedlinePgn>e28479</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1371/journal.pone.0028479</ELocationID><Abstract><AbstractText>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.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Frieman</LastName><ForeName>Matthew</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>Department of Microbiology and Immunology, University of Maryland, Baltimore, Maryland, United States of America.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Basu</LastName><ForeName>Dipanwita</ForeName><Initials>D</Initials></Author><Author ValidYN="Y"><LastName>Matthews</LastName><ForeName>Krystal</ForeName><Initials>K</Initials></Author><Author ValidYN="Y"><LastName>Taylor</LastName><ForeName>Justin</ForeName><Initials>J</Initials></Author><Author ValidYN="Y"><LastName>Jones</LastName><ForeName>Grant</ForeName><Initials>G</Initials></Author><Author ValidYN="Y"><LastName>Pickles</LastName><ForeName>Raymond</ForeName><Initials>R</Initials></Author><Author ValidYN="Y"><LastName>Baric</LastName><ForeName>Ralph</ForeName><Initials>R</Initials></Author><Author ValidYN="Y"><LastName>Engel</LastName><ForeName>Daniel A</ForeName><Initials>DA</Initials></Author></AuthorList><Language>eng</Language><GrantList CompleteYN="Y"><Grant><GrantID>K22 AI077797</GrantID><Acronym>AI</Acronym><Agency>NIAID NIH HHS</Agency><Country>United States</Country></Grant><Grant><GrantID>R21 NS063854</GrantID><Acronym>NS</Acronym><Agency>NINDS NIH HHS</Agency><Country>United States</Country></Grant><Grant><GrantID>1R21NS063854</GrantID><Acronym>NS</Acronym><Agency>NINDS NIH HHS</Agency><Country>United States</Country></Grant><Grant><GrantID>K22AI077797</GrantID><Acronym>AI</Acronym><Agency>NIAID NIH HHS</Agency><Country>United 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