The Nucleocapsid Protein of SARS-CoV Induces Transcription of hfgl2 Prothrombinase Gene Dependent on C/EBP Alpha
Identifieur interne : 001108 ( Pmc/Curation ); précédent : 001107; suivant : 001109The Nucleocapsid Protein of SARS-CoV Induces Transcription of hfgl2 Prothrombinase Gene Dependent on C/EBP Alpha
Auteurs : Meifang Han ; Weiming Yan ; Yuancheng Huang ; Huaning Yao ; Zhanhui Wang ; Dong Xi ; Weina Li ; Yaoyong Zhou ; Jinlin Hou ; Xiaoping Luo ; Qin NingSource :
- 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
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DOI: 10.1093/jb/mvn042
PubMed: 18390877
PubMed Central: 7109852
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<sourceDesc><biblStruct><analytic><title xml:lang="en" 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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<series><title level="j">Journal of Biochemistry</title>
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<front><div type="abstract" xml:lang="en"><title>Abstract</title>
<p>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 <italic>cis</italic>
-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.</p>
</div>
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<pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">J Biochem</journal-id>
<journal-id journal-id-type="iso-abbrev">J. Biochem</journal-id>
<journal-id journal-id-type="hwp">jbiochem</journal-id>
<journal-id journal-id-type="publisher-id">jbchem</journal-id>
<journal-title-group><journal-title>Journal of Biochemistry</journal-title>
</journal-title-group>
<issn pub-type="ppub">0021-924X</issn>
<issn pub-type="epub">1756-2651</issn>
<publisher><publisher-name>Oxford University Press</publisher-name>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">18390877</article-id>
<article-id pub-id-type="pmc">7109852</article-id>
<article-id pub-id-type="doi">10.1093/jb/mvn042</article-id>
<article-id pub-id-type="publisher-id">mvn042</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Regular Papers</subject>
</subj-group>
</article-categories>
<title-group><article-title>The Nucleocapsid Protein of SARS-CoV Induces Transcription of hfgl2 Prothrombinase Gene Dependent on C/EBP Alpha</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Han</surname>
<given-names>Meifang</given-names>
</name>
<xref ref-type="aff" rid="AFF1">1</xref>
<xref ref-type="aff" rid="AFF1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Yan</surname>
<given-names>Weiming</given-names>
</name>
<xref ref-type="aff" rid="AFF1">1</xref>
<xref ref-type="aff" rid="AFF1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Huang</surname>
<given-names>Yuancheng</given-names>
</name>
<xref ref-type="aff" rid="AFF1">1</xref>
<xref ref-type="aff" rid="AFF1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Yao</surname>
<given-names>Huaning</given-names>
</name>
<xref ref-type="aff" rid="AFF1">1</xref>
<xref ref-type="aff" rid="AFF1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Wang</surname>
<given-names>Zhanhui</given-names>
</name>
<xref ref-type="aff" rid="AFF1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Xi</surname>
<given-names>Dong</given-names>
</name>
<xref ref-type="aff" rid="AFF1">1</xref>
<xref ref-type="aff" rid="AFF1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Li</surname>
<given-names>Weina</given-names>
</name>
<xref ref-type="aff" rid="AFF1">1</xref>
<xref ref-type="aff" rid="AFF1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Zhou</surname>
<given-names>Yaoyong</given-names>
</name>
<xref ref-type="aff" rid="AFF1">1</xref>
<xref ref-type="aff" rid="AFF1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Hou</surname>
<given-names>Jinlin</given-names>
</name>
<xref ref-type="aff" rid="AFF1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Luo</surname>
<given-names>Xiaoping</given-names>
</name>
<xref ref-type="aff" rid="AFF1">1</xref>
<xref ref-type="aff" rid="AFF1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Ning</surname>
<given-names>Qin</given-names>
</name>
<xref ref-type="aff" rid="AFF1">1</xref>
<xref ref-type="aff" rid="AFF1">1</xref>
<xref ref-type="corresp" rid="COR1">*</xref>
<pmc-comment>qning@tjh.tjmu.edu.cn </pmc-comment>
</contrib>
</contrib-group>
<aff id="AFF1"><label><sup>1</sup>
</label>
Department of Infectious Disease;<sup>2</sup>
Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030;<sup>3</sup>
Department of Infectious Disease, Nanfang Hospital, Southern Medical University, Guangzhou, 510515; and<sup>4</sup>
Department of Pediatrics, Huazhong University of Science and Technology, Wuhan, 430030, China</aff>
<author-notes><corresp id="COR1">*To whom correspondence should be addressed. Tel: <phone>+86 27 83662391</phone>
, Fax: <fax>+86 27 83662391</fax>
, E-mail: <email>qning@tjh.tjmu.edu.cn</email>
</corresp>
</author-notes>
<pub-date pub-type="ppub"><month>7</month>
<year>2008</year>
</pub-date>
<pub-date pub-type="epub" iso-8601-date="2008-04-04"><day>4</day>
<month>4</month>
<year>2008</year>
</pub-date>
<volume>144</volume>
<issue>1</issue>
<fpage>51</fpage>
<lpage>62</lpage>
<history><date date-type="received"><day>4</day>
<month>2</month>
<year>2008</year>
</date>
<date date-type="accepted"><day>11</day>
<month>3</month>
<year>2008</year>
</date>
</history>
<permissions><copyright-statement>© 2008 The Japanese Biochemical Society</copyright-statement>
<copyright-year>2008</copyright-year>
<license><license-p>This article is made available via the PMC Open Access Subset for unrestricted re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the COVID-19 pandemic or until permissions are revoked in writing. Upon expiration of these permissions, PMC is granted a perpetual license to make this article available via PMC and Europe PMC, consistent with existing copyright protections.</license-p>
</license>
</permissions>
<self-uri xlink:href="mvn042.pdf"></self-uri>
<abstract><title>Abstract</title>
<p>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 <italic>cis</italic>
-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.</p>
</abstract>
<kwd-group><kwd>gene regulation</kwd>
<kwd>hfgl2</kwd>
<kwd>nucleocapsid protein</kwd>
<kwd>prothrombinase</kwd>
<kwd>SARS-CoV</kwd>
</kwd-group>
</article-meta>
</front>
</pmc>
</record>
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