SARS-CoV nucleocapsid protein interacts with cellular pyruvate kinase protein and inhibits its activity
Identifieur interne : 000040 ( PascalFrancis/Checkpoint ); précédent : 000039; suivant : 000041SARS-CoV nucleocapsid protein interacts with cellular pyruvate kinase protein and inhibits its activity
Auteurs : Wei-Yen Wei [Taïwan] ; Hui-Chun Li [Taïwan] ; Chiung-Yao Chen [Taïwan] ; Chee-Hing Yang [Taïwan] ; Shen-Kao Lee [Taïwan] ; Chia-Wen Wang [Taïwan] ; Hsin-Chieh Ma [Taïwan] ; Yue-Li Juang [Taïwan] ; Shih-Yen Lo [Taïwan]Source :
- Archives of virology [ 0304-8608 ] ; 2012.
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Abstract
The pathogenesis of SARS-CoV remains largely unknown. To study the function of the SARS-CoV nucleocapsid protein, we have conducted a yeast two-hybrid screening experiment to identify cellular proteins that may interact with the SARS-CoV nucleocapsid protein. Pyruvate kinase (liver) was found to interact with SARS-CoV nucleocapsid protein in this experiment. The binding domains of these two proteins were also determined using the yeast two-hybrid system. The physical interaction between the SARS-CoV nucleocapsid and cellular pyruvate kinase (liver) proteins was further confirmed by GST pull-down assay, co-immunoprecipitation assay and confocal microscopy. Cellular pyruvate kinase activity in hepatoma cells was repressed by SARS-CoV nucleocapsid protein in either transiently transfected or stably transfected cells. PK deficiency in red blood cells is known to result in human hereditary non-spherocytic hemolytic anemia. It is reasonable to assume that an inhibition of PKL activity due to interaction with SARS-CoV N protein is likely to cause the death of the hepatocytes, which results in the elevation of serum alanine aminotransferase and liver dysfunction noted in most SARS patients. Thus, our results suggest that SARS-CoV could reduce pyruvate kinase activity via its nucleocapsid protein, and this may in turn cause disease.
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<front><div type="abstract" xml:lang="en">The pathogenesis of SARS-CoV remains largely unknown. To study the function of the SARS-CoV nucleocapsid protein, we have conducted a yeast two-hybrid screening experiment to identify cellular proteins that may interact with the SARS-CoV nucleocapsid protein. Pyruvate kinase (liver) was found to interact with SARS-CoV nucleocapsid protein in this experiment. The binding domains of these two proteins were also determined using the yeast two-hybrid system. The physical interaction between the SARS-CoV nucleocapsid and cellular pyruvate kinase (liver) proteins was further confirmed by GST pull-down assay, co-immunoprecipitation assay and confocal microscopy. Cellular pyruvate kinase activity in hepatoma cells was repressed by SARS-CoV nucleocapsid protein in either transiently transfected or stably transfected cells. PK deficiency in red blood cells is known to result in human hereditary non-spherocytic hemolytic anemia. It is reasonable to assume that an inhibition of PKL activity due to interaction with SARS-CoV N protein is likely to cause the death of the hepatocytes, which results in the elevation of serum alanine aminotransferase and liver dysfunction noted in most SARS patients. Thus, our results suggest that SARS-CoV could reduce pyruvate kinase activity via its nucleocapsid protein, and this may in turn cause disease.</div>
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<fA66 i1="01"><s0>AUT</s0>
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<fC01 i1="01" l="ENG"><s0>The pathogenesis of SARS-CoV remains largely unknown. To study the function of the SARS-CoV nucleocapsid protein, we have conducted a yeast two-hybrid screening experiment to identify cellular proteins that may interact with the SARS-CoV nucleocapsid protein. Pyruvate kinase (liver) was found to interact with SARS-CoV nucleocapsid protein in this experiment. The binding domains of these two proteins were also determined using the yeast two-hybrid system. The physical interaction between the SARS-CoV nucleocapsid and cellular pyruvate kinase (liver) proteins was further confirmed by GST pull-down assay, co-immunoprecipitation assay and confocal microscopy. Cellular pyruvate kinase activity in hepatoma cells was repressed by SARS-CoV nucleocapsid protein in either transiently transfected or stably transfected cells. PK deficiency in red blood cells is known to result in human hereditary non-spherocytic hemolytic anemia. It is reasonable to assume that an inhibition of PKL activity due to interaction with SARS-CoV N protein is likely to cause the death of the hepatocytes, which results in the elevation of serum alanine aminotransferase and liver dysfunction noted in most SARS patients. Thus, our results suggest that SARS-CoV could reduce pyruvate kinase activity via its nucleocapsid protein, and this may in turn cause disease.</s0>
</fC01>
<fC02 i1="01" i2="X"><s0>002A05C10</s0>
</fC02>
<fC03 i1="01" i2="X" l="FRE"><s0>Virus syndrome respiratoire aigu sévère</s0>
<s2>NW</s2>
<s5>01</s5>
</fC03>
<fC03 i1="01" i2="X" l="ENG"><s0>Severe acute respiratory syndrome virus</s0>
<s2>NW</s2>
<s5>01</s5>
</fC03>
<fC03 i1="01" i2="X" l="SPA"><s0>Severe acute respiratory syndrome virus</s0>
<s2>NW</s2>
<s5>01</s5>
</fC03>
<fC03 i1="02" i2="X" l="FRE"><s0>Nucléocapside</s0>
<s5>05</s5>
</fC03>
<fC03 i1="02" i2="X" l="ENG"><s0>Nucleocapsid</s0>
<s5>05</s5>
</fC03>
<fC03 i1="02" i2="X" l="SPA"><s0>Nucleocápside</s0>
<s5>05</s5>
</fC03>
<fC03 i1="03" i2="X" l="FRE"><s0>Non-specific serine/threonine protein kinase</s0>
<s2>FE</s2>
<s5>06</s5>
</fC03>
<fC03 i1="03" i2="X" l="ENG"><s0>Non-specific serine/threonine protein kinase</s0>
<s2>FE</s2>
<s5>06</s5>
</fC03>
<fC03 i1="03" i2="X" l="SPA"><s0>Non-specific serine/threonine protein kinase</s0>
<s2>FE</s2>
<s5>06</s5>
</fC03>
<fC03 i1="04" i2="X" l="FRE"><s0>Pyruvate kinase</s0>
<s2>FE</s2>
<s5>07</s5>
</fC03>
<fC03 i1="04" i2="X" l="ENG"><s0>Pyruvate kinase</s0>
<s2>FE</s2>
<s5>07</s5>
</fC03>
<fC03 i1="04" i2="X" l="SPA"><s0>Pyruvate kinase</s0>
<s2>FE</s2>
<s5>07</s5>
</fC03>
<fC07 i1="01" i2="X" l="FRE"><s0>Coronavirus</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="01" i2="X" l="ENG"><s0>Coronavirus</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="01" i2="X" l="SPA"><s0>Coronavirus</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="02" i2="X" l="FRE"><s0>Coronaviridae</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="02" i2="X" l="ENG"><s0>Coronaviridae</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="02" i2="X" l="SPA"><s0>Coronaviridae</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="03" i2="X" l="FRE"><s0>Nidovirales</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="03" i2="X" l="ENG"><s0>Nidovirales</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="03" i2="X" l="SPA"><s0>Nidovirales</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="04" i2="X" l="FRE"><s0>Virus</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="04" i2="X" l="ENG"><s0>Virus</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="04" i2="X" l="SPA"><s0>Virus</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="05" i2="X" l="FRE"><s0>Transferases</s0>
<s2>FE</s2>
</fC07>
<fC07 i1="05" i2="X" l="ENG"><s0>Transferases</s0>
<s2>FE</s2>
</fC07>
<fC07 i1="05" i2="X" l="SPA"><s0>Transferases</s0>
<s2>FE</s2>
</fC07>
<fC07 i1="06" i2="X" l="FRE"><s0>Enzyme</s0>
<s2>FE</s2>
</fC07>
<fC07 i1="06" i2="X" l="ENG"><s0>Enzyme</s0>
<s2>FE</s2>
</fC07>
<fC07 i1="06" i2="X" l="SPA"><s0>Enzima</s0>
<s2>FE</s2>
</fC07>
<fN21><s1>135</s1>
</fN21>
<fN44 i1="01"><s1>OTO</s1>
</fN44>
<fN82><s1>OTO</s1>
</fN82>
</pA>
</standard>
</inist>
<affiliations><list><country><li>Taïwan</li>
</country>
</list>
<tree><country name="Taïwan"><noRegion><name sortKey="Wei, Wei Yen" sort="Wei, Wei Yen" uniqKey="Wei W" first="Wei-Yen" last="Wei">Wei-Yen Wei</name>
</noRegion>
<name sortKey="Chen, Chiung Yao" sort="Chen, Chiung Yao" uniqKey="Chen C" first="Chiung-Yao" last="Chen">Chiung-Yao Chen</name>
<name sortKey="Juang, Yue Li" sort="Juang, Yue Li" uniqKey="Juang Y" first="Yue-Li" last="Juang">Yue-Li Juang</name>
<name sortKey="Juang, Yue Li" sort="Juang, Yue Li" uniqKey="Juang Y" first="Yue-Li" last="Juang">Yue-Li Juang</name>
<name sortKey="Lee, Shen Kao" sort="Lee, Shen Kao" uniqKey="Lee S" first="Shen-Kao" last="Lee">Shen-Kao Lee</name>
<name sortKey="Li, Hui Chun" sort="Li, Hui Chun" uniqKey="Li H" first="Hui-Chun" last="Li">Hui-Chun Li</name>
<name sortKey="Li, Hui Chun" sort="Li, Hui Chun" uniqKey="Li H" first="Hui-Chun" last="Li">Hui-Chun Li</name>
<name sortKey="Lo, Shih Yen" sort="Lo, Shih Yen" uniqKey="Lo S" first="Shih-Yen" last="Lo">Shih-Yen Lo</name>
<name sortKey="Lo, Shih Yen" sort="Lo, Shih Yen" uniqKey="Lo S" first="Shih-Yen" last="Lo">Shih-Yen Lo</name>
<name sortKey="Lo, Shih Yen" sort="Lo, Shih Yen" uniqKey="Lo S" first="Shih-Yen" last="Lo">Shih-Yen Lo</name>
<name sortKey="Ma, Hsin Chieh" sort="Ma, Hsin Chieh" uniqKey="Ma H" first="Hsin-Chieh" last="Ma">Hsin-Chieh Ma</name>
<name sortKey="Wang, Chia Wen" sort="Wang, Chia Wen" uniqKey="Wang C" first="Chia-Wen" last="Wang">Chia-Wen Wang</name>
<name sortKey="Yang, Chee Hing" sort="Yang, Chee Hing" uniqKey="Yang C" first="Chee-Hing" last="Yang">Chee-Hing Yang</name>
</country>
</tree>
</affiliations>
</record>
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