Yeast-based assays for the high-throughput screening of inhibitors of coronavirus RNA cap guanine-N7-methyltransferase
Identifieur interne : 000974 ( PascalFrancis/Curation ); précédent : 000973; suivant : 000975Yeast-based assays for the high-throughput screening of inhibitors of coronavirus RNA cap guanine-N7-methyltransferase
Auteurs : YING SUN [République populaire de Chine] ; ZIDAO WANG [République populaire de Chine] ; JIALI TAO [République populaire de Chine] ; YI WANG [République populaire de Chine] ; ANDONG WU [République populaire de Chine] ; ZIWEN YANG [République populaire de Chine] ; KAIMEI WANG [République populaire de Chine] ; LIQIAO SHI [République populaire de Chine] ; YU CHEN [République populaire de Chine] ; DEYIN GUO [République populaire de Chine]Source :
- Antiviral research [ 0166-3542 ] ; 2014.
Descripteurs français
- Pascal (Inist)
- Wicri :
- topic : Antibiotique.
English descriptors
- KwdEn :
Abstract
The 5?-cap structure is a distinct feature of eukaryotic mRNAs and is important for RNA stability and protein translation by providing a molecular signature for the distinction of self or non-self mRNA. Eukaryotic viruses generally modify the 5?-end of their RNAs to mimic the cellular mRNA structure, thereby facilitating viral replication in host cells. However, the molecular organization and biochemical mechanisms of the viral capping apparatus typically differ from its cellular counterpart, which makes viral capping enzymes attractive targets for drug discovery. Our previous work showed that SARS coronavirus (SARS-CoV) non-structural protein 14 represents a structurally novel and unique guanine-N7-methyl-transferase (N7-MTase) that is able to functionally complement yeast cellular N7-MTase. In the present study, we developed a yeast-based system for identifying and screening inhibitors against coronavirus N7-MTase using both 96-well and 384-well microtiter plates. The MTase inhibitors previously identified by in vitro biochemical assays were tested, and some, such as sinefungin, effectively suppressed N7-MTase in the yeast system. However, other compounds, such as ATA and AdoHcy, did not exert an inhibitory effect within a cellular context. These results validated the yeast assay system for inhibitor screening yet also demonstrated the difference between cell-based and in vitro biochemical assays. The yeast system was applied to the screening of 3000 natural product extracts, and three were observed to more potently inhibit the activity of coronavirus than human N7-MTase.
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430072</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>République populaire de Chine</country></affiliation></author><author><name sortKey="Deyin Guo" sort="Deyin Guo" uniqKey="Deyin Guo" last="Deyin Guo">DEYIN GUO</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>State Key Laboratory of Virology, College of Life Sciences, Wuhan University</s1><s2>Wuhan 430072</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>République populaire de Chine</country></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">INIST</idno><idno type="inist">14-0104920</idno><date when="2014">2014</date><idno type="stanalyst">PASCAL 14-0104920 INIST</idno><idno type="RBID">Pascal:14-0104920</idno><idno type="wicri:Area/PascalFrancis/Corpus">000013</idno><idno type="wicri:Area/PascalFrancis/Curation">000974</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">Yeast-based assays for the high-throughput screening of inhibitors of coronavirus RNA cap guanine-N7-methyltransferase</title><author><name sortKey="Ying Sun" sort="Ying Sun" uniqKey="Ying Sun" last="Ying Sun">YING SUN</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>State Key Laboratory of Virology, College of Life Sciences, Wuhan University</s1><s2>Wuhan 430072</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>République populaire de Chine</country></affiliation></author><author><name sortKey="Zidao Wang" sort="Zidao Wang" uniqKey="Zidao Wang" last="Zidao Wang">ZIDAO WANG</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>State Key Laboratory of Virology, College of Life Sciences, Wuhan University</s1><s2>Wuhan 430072</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>République populaire de Chine</country></affiliation></author><author><name sortKey="Jiali Tao" sort="Jiali Tao" uniqKey="Jiali Tao" last="Jiali Tao">JIALI TAO</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>State Key Laboratory of Virology, College of Life Sciences, Wuhan University</s1><s2>Wuhan 430072</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>République populaire de Chine</country></affiliation></author><author><name sortKey="Yi Wang" sort="Yi Wang" uniqKey="Yi Wang" last="Yi Wang">YI WANG</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>State Key Laboratory of Virology, College of Life Sciences, Wuhan University</s1><s2>Wuhan 430072</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>République populaire de Chine</country></affiliation></author><author><name sortKey="Andong Wu" sort="Andong Wu" uniqKey="Andong Wu" last="Andong Wu">ANDONG WU</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>State Key Laboratory of Virology, College of Life Sciences, Wuhan University</s1><s2>Wuhan 430072</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>République populaire de Chine</country></affiliation></author><author><name sortKey="Ziwen Yang" sort="Ziwen Yang" uniqKey="Ziwen Yang" last="Ziwen Yang">ZIWEN YANG</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Biological Pesticide Engineering Research Center, Hubei Academy of Agricultural Science</s1><s2>Hubei 430072</s2><s3>CHN</s3><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>République populaire de Chine</country></affiliation></author><author><name sortKey="Kaimei Wang" sort="Kaimei Wang" uniqKey="Kaimei Wang" last="Kaimei Wang">KAIMEI WANG</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Biological Pesticide Engineering Research Center, Hubei Academy of Agricultural Science</s1><s2>Hubei 430072</s2><s3>CHN</s3><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>République populaire de Chine</country></affiliation></author><author><name sortKey="Liqiao Shi" sort="Liqiao Shi" uniqKey="Liqiao Shi" last="Liqiao Shi">LIQIAO SHI</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Biological Pesticide Engineering Research Center, Hubei Academy of Agricultural Science</s1><s2>Hubei 430072</s2><s3>CHN</s3><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>République populaire de Chine</country></affiliation></author><author><name sortKey="Yu Chen" sort="Yu Chen" uniqKey="Yu Chen" last="Yu Chen">YU CHEN</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>State Key Laboratory of Virology, College of Life Sciences, Wuhan University</s1><s2>Wuhan 430072</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>République populaire de Chine</country></affiliation></author><author><name sortKey="Deyin Guo" sort="Deyin Guo" uniqKey="Deyin Guo" last="Deyin Guo">DEYIN GUO</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>State Key Laboratory of Virology, College of Life Sciences, Wuhan University</s1><s2>Wuhan 430072</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>République populaire de Chine</country></affiliation></author></analytic><series><title level="j" type="main">Antiviral research</title><title level="j" type="abbreviated">Antivir. res.</title><idno type="ISSN">0166-3542</idno><imprint><date when="2014">2014</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Antiviral research</title><title level="j" type="abbreviated">Antivir. res.</title><idno type="ISSN">0166-3542</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antibiotic</term><term>Coronavirus</term><term>High throughput screening</term><term>Inhibitor</term><term>Methyltransferases</term><term>Parasiticide</term><term>Sinefungin</term><term>Yeast</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Levure</term><term>Criblage haut débit</term><term>Inhibiteur</term><term>Coronavirus</term><term>Methyltransferases</term><term>Sinéfungine</term><term>Antibiotique</term><term>Antiparasitaire</term><term>Coiffe ARN</term></keywords><keywords scheme="Wicri" type="topic" xml:lang="fr"><term>Antibiotique</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The 5?-cap structure is a distinct feature of eukaryotic mRNAs and is important for RNA stability and protein translation by providing a molecular signature for the distinction of self or non-self mRNA. Eukaryotic viruses generally modify the 5?-end of their RNAs to mimic the cellular mRNA structure, thereby facilitating viral replication in host cells. However, the molecular organization and biochemical mechanisms of the viral capping apparatus typically differ from its cellular counterpart, which makes viral capping enzymes attractive targets for drug discovery. Our previous work showed that SARS coronavirus (SARS-CoV) non-structural protein 14 represents a structurally novel and unique guanine-N7-methyl-transferase (N7-MTase) that is able to functionally complement yeast cellular N7-MTase. In the present study, we developed a yeast-based system for identifying and screening inhibitors against coronavirus N7-MTase using both 96-well and 384-well microtiter plates. The MTase inhibitors previously identified by in vitro biochemical assays were tested, and some, such as sinefungin, effectively suppressed N7-MTase in the yeast system. However, other compounds, such as ATA and AdoHcy, did not exert an inhibitory effect within a cellular context. These results validated the yeast assay system for inhibitor screening yet also demonstrated the difference between cell-based and in vitro biochemical assays. The yeast system was applied to the screening of 3000 natural product extracts, and three were observed to more potently inhibit the activity of coronavirus than human N7-MTase.</div></front></TEI><inist><standard h6="B"><pA><fA01 i1="01" i2="1"><s0>0166-3542</s0></fA01><fA02 i1="01"><s0>ARSRDR</s0></fA02><fA03 i2="1"><s0>Antivir. res.</s0></fA03><fA05><s2>104</s2></fA05><fA08 i1="01" i2="1" l="ENG"><s1>Yeast-based assays for the high-throughput screening of inhibitors of coronavirus RNA cap guanine-N7-methyltransferase</s1></fA08><fA11 i1="01" i2="1"><s1>YING SUN</s1></fA11><fA11 i1="02" i2="1"><s1>ZIDAO WANG</s1></fA11><fA11 i1="03" i2="1"><s1>JIALI TAO</s1></fA11><fA11 i1="04" i2="1"><s1>YI WANG</s1></fA11><fA11 i1="05" i2="1"><s1>ANDONG WU</s1></fA11><fA11 i1="06" i2="1"><s1>ZIWEN YANG</s1></fA11><fA11 i1="07" i2="1"><s1>KAIMEI WANG</s1></fA11><fA11 i1="08" i2="1"><s1>LIQIAO SHI</s1></fA11><fA11 i1="09" i2="1"><s1>YU CHEN</s1></fA11><fA11 i1="10" i2="1"><s1>DEYIN GUO</s1></fA11><fA14 i1="01"><s1>State Key Laboratory of Virology, College of Life Sciences, Wuhan University</s1><s2>Wuhan 430072</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ></fA14><fA14 i1="02"><s1>Biological Pesticide Engineering Research Center, Hubei Academy of Agricultural Science</s1><s2>Hubei 430072</s2><s3>CHN</s3><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></fA14><fA20><s1>156-164</s1></fA20><fA21><s1>2014</s1></fA21><fA23 i1="01"><s0>ENG</s0></fA23><fA43 i1="01"><s1>INIST</s1><s2>18839</s2><s5>354000506165990200</s5></fA43><fA44><s0>0000</s0><s1>© 2014 INIST-CNRS. All rights reserved.</s1></fA44><fA45><s0>1 p.</s0></fA45><fA47 i1="01" i2="1"><s0>14-0104920</s0></fA47><fA60><s1>P</s1></fA60><fA61><s0>A</s0></fA61><fA64 i1="01" i2="1"><s0>Antiviral research</s0></fA64><fA66 i1="01"><s0>GBR</s0></fA66><fC01 i1="01" l="ENG"><s0>The 5?-cap structure is a distinct feature of eukaryotic mRNAs and is important for RNA stability and protein translation by providing a molecular signature for the distinction of self or non-self mRNA. Eukaryotic viruses generally modify the 5?-end of their RNAs to mimic the cellular mRNA structure, thereby facilitating viral replication in host cells. However, the molecular organization and biochemical mechanisms of the viral capping apparatus typically differ from its cellular counterpart, which makes viral capping enzymes attractive targets for drug discovery. Our previous work showed that SARS coronavirus (SARS-CoV) non-structural protein 14 represents a structurally novel and unique guanine-N7-methyl-transferase (N7-MTase) that is able to functionally complement yeast cellular N7-MTase. In the present study, we developed a yeast-based system for identifying and screening inhibitors against coronavirus N7-MTase using both 96-well and 384-well microtiter plates. The MTase inhibitors previously identified by in vitro biochemical assays were tested, and some, such as sinefungin, effectively suppressed N7-MTase in the yeast system. However, other compounds, such as ATA and AdoHcy, did not exert an inhibitory effect within a cellular context. These results validated the yeast assay system for inhibitor screening yet also demonstrated the difference between cell-based and in vitro biochemical assays. The yeast system was applied to the screening of 3000 natural product extracts, and three were observed to more potently inhibit the activity of coronavirus than human N7-MTase.</s0></fC01><fC02 i1="01" i2="X"><s0>002B02S05</s0></fC02><fC03 i1="01" i2="X" l="FRE"><s0>Levure</s0><s5>01</s5></fC03><fC03 i1="01" i2="X" l="ENG"><s0>Yeast</s0><s5>01</s5></fC03><fC03 i1="01" i2="X" l="SPA"><s0>Levadura</s0><s5>01</s5></fC03><fC03 i1="02" i2="X" l="FRE"><s0>Criblage haut débit</s0><s5>02</s5></fC03><fC03 i1="02" i2="X" l="ENG"><s0>High throughput screening</s0><s5>02</s5></fC03><fC03 i1="02" i2="X" l="SPA"><s0>Cribado alta productividad</s0><s5>02</s5></fC03><fC03 i1="03" i2="X" l="FRE"><s0>Inhibiteur</s0><s5>03</s5></fC03><fC03 i1="03" i2="X" l="ENG"><s0>Inhibitor</s0><s5>03</s5></fC03><fC03 i1="03" i2="X" l="SPA"><s0>Inhibidor</s0><s5>03</s5></fC03><fC03 i1="04" i2="X" l="FRE"><s0>Coronavirus</s0><s2>NW</s2><s5>04</s5></fC03><fC03 i1="04" i2="X" l="ENG"><s0>Coronavirus</s0><s2>NW</s2><s5>04</s5></fC03><fC03 i1="04" i2="X" l="SPA"><s0>Coronavirus</s0><s2>NW</s2><s5>04</s5></fC03><fC03 i1="05" i2="X" l="FRE"><s0>Methyltransferases</s0><s2>FE</s2><s5>05</s5></fC03><fC03 i1="05" i2="X" l="ENG"><s0>Methyltransferases</s0><s2>FE</s2><s5>05</s5></fC03><fC03 i1="05" i2="X" l="SPA"><s0>Methyltransferases</s0><s2>FE</s2><s5>05</s5></fC03><fC03 i1="06" i2="X" l="FRE"><s0>Sinéfungine</s0><s2>NK</s2><s2>FR</s2><s5>06</s5></fC03><fC03 i1="06" i2="X" l="ENG"><s0>Sinefungin</s0><s2>NK</s2><s2>FR</s2><s5>06</s5></fC03><fC03 i1="06" i2="X" l="SPA"><s0>Sinefungina</s0><s2>NK</s2><s2>FR</s2><s5>06</s5></fC03><fC03 i1="07" i2="X" l="FRE"><s0>Antibiotique</s0><s5>23</s5></fC03><fC03 i1="07" i2="X" l="ENG"><s0>Antibiotic</s0><s5>23</s5></fC03><fC03 i1="07" i2="X" l="SPA"><s0>Antibiótico</s0><s5>23</s5></fC03><fC03 i1="08" i2="X" l="FRE"><s0>Antiparasitaire</s0><s5>24</s5></fC03><fC03 i1="08" i2="X" l="ENG"><s0>Parasiticide</s0><s5>24</s5></fC03><fC03 i1="08" i2="X" l="SPA"><s0>Antiparasitario</s0><s5>24</s5></fC03><fC03 i1="09" i2="X" l="FRE"><s0>Coiffe ARN</s0><s4>INC</s4><s5>86</s5></fC03><fC07 i1="01" i2="X" l="FRE"><s0>Coronaviridae</s0><s2>NW</s2></fC07><fC07 i1="01" i2="X" l="ENG"><s0>Coronaviridae</s0><s2>NW</s2></fC07><fC07 i1="01" i2="X" l="SPA"><s0>Coronaviridae</s0><s2>NW</s2></fC07><fC07 i1="02" i2="X" l="FRE"><s0>Nidovirales</s0><s2>NW</s2></fC07><fC07 i1="02" i2="X" l="ENG"><s0>Nidovirales</s0><s2>NW</s2></fC07><fC07 i1="02" i2="X" l="SPA"><s0>Nidovirales</s0><s2>NW</s2></fC07><fC07 i1="03" i2="X" l="FRE"><s0>Virus</s0><s2>NW</s2></fC07><fC07 i1="03" i2="X" l="ENG"><s0>Virus</s0><s2>NW</s2></fC07><fC07 i1="03" i2="X" l="SPA"><s0>Virus</s0><s2>NW</s2></fC07><fC07 i1="04" i2="X" l="FRE"><s0>Transferases</s0><s2>FE</s2></fC07><fC07 i1="04" i2="X" l="ENG"><s0>Transferases</s0><s2>FE</s2></fC07><fC07 i1="04" i2="X" l="SPA"><s0>Transferases</s0><s2>FE</s2></fC07><fC07 i1="05" i2="X" l="FRE"><s0>Enzyme</s0><s2>FE</s2></fC07><fC07 i1="05" i2="X" l="ENG"><s0>Enzyme</s0><s2>FE</s2></fC07><fC07 i1="05" i2="X" l="SPA"><s0>Enzima</s0><s2>FE</s2></fC07><fN21><s1>139</s1></fN21><fN44 i1="01"><s1>OTO</s1></fN44><fN82><s1>OTO</s1></fN82></pA></standard></inist></record>Pour manipuler ce document sous Unix (Dilib)
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|wiki= Sante
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|texte= Yeast-based assays for the high-throughput screening of inhibitors of coronavirus RNA cap guanine-N7-methyltransferase
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