The Nucleocapsid Protein of SARS-CoV Induces Transcription of hfgl2 Prothrombinase Gene Dependent on C/EBP Alpha
Identifieur interne : 000339 ( Istex/Curation ); précédent : 000338; suivant : 000340The Nucleocapsid Protein of SARS-CoV Induces Transcription of hfgl2 Prothrombinase Gene Dependent on C/EBP Alpha
Auteurs : Meifang Han [République populaire de Chine] ; Weiming Yan [République populaire de Chine] ; Yuancheng Huang [République populaire de Chine] ; Huaning Yao [République populaire de Chine] ; Zhanhui Wang [République populaire de Chine] ; Dong Xi [République populaire de Chine] ; Weina Li [République populaire de Chine] ; Yaoyong Zhou [République populaire de Chine] ; Jinlin Hou [République populaire de Chine] ; Xiaoping Luo [République populaire de Chine] ; Qin Ning [République populaire de Chine]Source :
- Journal of Biochemistry [ 0021-924X ] ; 2008.
Abstract
Fibrin deposition was universal in the lungs of SARS patients and fgl2 prothrombinase gene, a novel procoagulant, was demonstrated to express highly in a clinically relevant SARS model. To investigate whether and which structural protein of SARS-CoV induced transcription of hfgl2 prothrombinase gene, three eukaryotic expression plasmids expressing nucleocapsid protein (N), membrane protein (M) and spike protein 2 (S2) of SARS-CoV were co-transfected with hfgl2 promoter luciferase-reporter plasmids and β-galactosidase plasmid in CHO cells, respectively. M, N and S2 protein of SARS-CoV were detected by western blotting and immunohistochemistry analysis. Further assays demonstrated that expression of hfgl2 gene was related with N protein, but not with M or S2 protein in THP-1 cells and Vero cells. N protein significantly induced functional procoagulant activity in comparison with control group. Luciferase assay showed that N protein of SARS-CoV could activate the transcription of hfgl2 promoter compared with the pcDNA3.1 empty vector. Site-directed mutagenesis and EMSA assay further demonstrated that transcription factor C/EBP alpha band with its cognate cis-element in hfgl2 promoter. The results showed that N protein of SARS-CoV induced hfgl2 gene transcription dependent on the transcription factor C/EBP alpha, which maybe contribute to the development of thrombosis in SARS.
Url:
DOI: 10.1093/jb/mvn042
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<sourceDesc><biblStruct><analytic><title level="a" type="main">The Nucleocapsid Protein of SARS-CoV Induces Transcription of hfgl2 Prothrombinase Gene Dependent on C/EBP Alpha</title>
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<author><name sortKey="Huang, Yuancheng" sort="Huang, Yuancheng" uniqKey="Huang Y" first="Yuancheng" last="Huang">Yuancheng Huang</name>
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<wicri:regionArea>Department of Infectious Disease; Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030; Department of Infectious Disease, Nanfang Hospital, Southern Medical University, Guangzhou, 510515; and Department of Pediatrics, Huazhong University of Science and Technology, Wuhan, 430030</wicri:regionArea>
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<author><name sortKey="Yao, Huaning" sort="Yao, Huaning" uniqKey="Yao H" first="Huaning" last="Yao">Huaning Yao</name>
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<wicri:regionArea>Department of Infectious Disease; Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030; Department of Infectious Disease, Nanfang Hospital, Southern Medical University, Guangzhou, 510515; and Department of Pediatrics, Huazhong University of Science and Technology, Wuhan, 430030</wicri:regionArea>
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<author><name sortKey="Wang, Zhanhui" sort="Wang, Zhanhui" uniqKey="Wang Z" first="Zhanhui" last="Wang">Zhanhui Wang</name>
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<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>Department of Infectious Disease; Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030; Department of Infectious Disease, Nanfang Hospital, Southern Medical University, Guangzhou, 510515; and Department of Pediatrics, Huazhong University of Science and Technology, Wuhan, 430030</wicri:regionArea>
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<author><name sortKey="Xi, Dong" sort="Xi, Dong" uniqKey="Xi D" first="Dong" last="Xi">Dong Xi</name>
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<author><name sortKey="Li, Weina" sort="Li, Weina" uniqKey="Li W" first="Weina" last="Li">Weina Li</name>
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<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>Department of Infectious Disease; Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030; Department of Infectious Disease, Nanfang Hospital, Southern Medical University, Guangzhou, 510515; and Department of Pediatrics, Huazhong University of Science and Technology, Wuhan, 430030</wicri:regionArea>
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<author><name sortKey="Zhou, Yaoyong" sort="Zhou, Yaoyong" uniqKey="Zhou Y" first="Yaoyong" last="Zhou">Yaoyong Zhou</name>
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<country xml:lang="fr">République populaire de Chine</country>
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<author><name sortKey="Hou, Jinlin" sort="Hou, Jinlin" uniqKey="Hou J" first="Jinlin" last="Hou">Jinlin Hou</name>
<affiliation wicri:level="1"><mods:affiliation>Department of Infectious Disease; Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030; Department of Infectious Disease, Nanfang Hospital, Southern Medical University, Guangzhou, 510515; and Department of Pediatrics, Huazhong University of Science and Technology, Wuhan, 430030, China</mods:affiliation>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>Department of Infectious Disease; Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030; Department of Infectious Disease, Nanfang Hospital, Southern Medical University, Guangzhou, 510515; and Department of Pediatrics, Huazhong University of Science and Technology, Wuhan, 430030</wicri:regionArea>
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<author><name sortKey="Luo, Xiaoping" sort="Luo, Xiaoping" uniqKey="Luo X" first="Xiaoping" last="Luo">Xiaoping Luo</name>
<affiliation wicri:level="1"><mods:affiliation>Department of Infectious Disease; Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030; Department of Infectious Disease, Nanfang Hospital, Southern Medical University, Guangzhou, 510515; and Department of Pediatrics, Huazhong University of Science and Technology, Wuhan, 430030, China</mods:affiliation>
<country xml:lang="fr">République populaire de Chine</country>
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<author><name sortKey="Ning, Qin" sort="Ning, Qin" uniqKey="Ning Q" first="Qin" last="Ning">Qin Ning</name>
<affiliation wicri:level="1"><mods:affiliation>Department of Infectious Disease; Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030; Department of Infectious Disease, Nanfang Hospital, Southern Medical University, Guangzhou, 510515; and Department of Pediatrics, Huazhong University of Science and Technology, Wuhan, 430030, China</mods:affiliation>
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<wicri:regionArea>Department of Infectious Disease; Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030; Department of Infectious Disease, Nanfang Hospital, Southern Medical University, Guangzhou, 510515; and Department of Pediatrics, Huazhong University of Science and Technology, Wuhan, 430030</wicri:regionArea>
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<country wicri:rule="url">République populaire de Chine</country>
</affiliation>
<affiliation wicri:level="1"><mods:affiliation>*To whom correspondence should be addressed. Tel: +86 27 83662391, Fax: +86 27 83662391, E-mail: qning@tjh.tjmu.edu.cn</mods:affiliation>
<country wicri:rule="url">République populaire de Chine</country>
<wicri:regionArea>*To whom correspondence should be addressed. Tel: +86 27 83662391, Fax: +86 27 83662391</wicri:regionArea>
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<front><div type="abstract">Fibrin deposition was universal in the lungs of SARS patients and fgl2 prothrombinase gene, a novel procoagulant, was demonstrated to express highly in a clinically relevant SARS model. To investigate whether and which structural protein of SARS-CoV induced transcription of hfgl2 prothrombinase gene, three eukaryotic expression plasmids expressing nucleocapsid protein (N), membrane protein (M) and spike protein 2 (S2) of SARS-CoV were co-transfected with hfgl2 promoter luciferase-reporter plasmids and β-galactosidase plasmid in CHO cells, respectively. M, N and S2 protein of SARS-CoV were detected by western blotting and immunohistochemistry analysis. Further assays demonstrated that expression of hfgl2 gene was related with N protein, but not with M or S2 protein in THP-1 cells and Vero cells. N protein significantly induced functional procoagulant activity in comparison with control group. Luciferase assay showed that N protein of SARS-CoV could activate the transcription of hfgl2 promoter compared with the pcDNA3.1 empty vector. Site-directed mutagenesis and EMSA assay further demonstrated that transcription factor C/EBP alpha band with its cognate cis-element in hfgl2 promoter. The results showed that N protein of SARS-CoV induced hfgl2 gene transcription dependent on the transcription factor C/EBP alpha, which maybe contribute to the development of thrombosis in SARS.</div>
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