Nucleocapsid protein of SARS-CoV activates the expression of cyclooxygenase-2 by binding directly to regulatory elements for nuclear factor-kappa B and CCAAT/enhancer binding protein.
Identifieur interne : 003D99 ( Main/Exploration ); précédent : 003D98; suivant : 003E00Nucleocapsid protein of SARS-CoV activates the expression of cyclooxygenase-2 by binding directly to regulatory elements for nuclear factor-kappa B and CCAAT/enhancer binding protein.
Auteurs : Xiaohong Yan [République populaire de Chine] ; Qian Hao ; Yongxin Mu ; Khalid Amine Timani ; Linbai Ye ; Ying Zhu ; Jianguo WuSource :
- The international journal of biochemistry & cell biology [ 1357-2725 ] ; 2006.
Descripteurs français
- KwdFr :
- Activation de la transcription, Cyclooxygenase 2 (génétique), Cyclooxygenase 2 (métabolisme), Facteur de transcription NF-kappa B (métabolisme), Humains, Immunoprécipitation de la chromatine, Liaison aux protéines, Lignée cellulaire, Mutagenèse dirigée (), Mutation, Plasmides (génétique), Protéine bêta de liaison aux séquences stimulatrices de type CCAAT (métabolisme), Protéines nucléocapside (génétique), Protéines nucléocapside (métabolisme), Protéines nucléocapside (physiologie), Régions promotrices (génétique) (génétique), Régulation de l'expression des gènes codant pour des enzymes, Sites de fixation (génétique), Séquence d'acides aminés, Test de retard de migration électrophorétique, Virus du SRAS (génétique), Virus du SRAS (métabolisme).
- MESH :
- génétique : Cyclooxygenase 2, Plasmides, Protéines nucléocapside, Régions promotrices (génétique), Sites de fixation, Virus du SRAS.
- métabolisme : Cyclooxygenase 2, Facteur de transcription NF-kappa B, Protéine bêta de liaison aux séquences stimulatrices de type CCAAT, Protéines nucléocapside, Virus du SRAS.
- physiologie : Protéines nucléocapside.
- Activation de la transcription, Humains, Immunoprécipitation de la chromatine, Liaison aux protéines, Lignée cellulaire, Mutagenèse dirigée, Mutation, Régulation de l'expression des gènes codant pour des enzymes, Séquence d'acides aminés, Test de retard de migration électrophorétique.
English descriptors
- KwdEn :
- Amino Acid Sequence, Binding Sites (genetics), CCAAT-Enhancer-Binding Protein-beta (metabolism), Cell Line, Chromatin Immunoprecipitation, Cyclooxygenase 2 (genetics), Cyclooxygenase 2 (metabolism), Electrophoretic Mobility Shift Assay, Gene Expression Regulation, Enzymologic, Humans, Mutagenesis, Site-Directed (methods), Mutation, NF-kappa B (metabolism), Nucleocapsid Proteins (genetics), Nucleocapsid Proteins (metabolism), Nucleocapsid Proteins (physiology), Plasmids (genetics), Promoter Regions, Genetic (genetics), Protein Binding, SARS Virus (genetics), SARS Virus (metabolism), Transcriptional Activation.
- MESH :
- chemical , genetics : Cyclooxygenase 2, Nucleocapsid Proteins.
- chemical , metabolism : CCAAT-Enhancer-Binding Protein-beta, Cyclooxygenase 2, NF-kappa B, Nucleocapsid Proteins.
- genetics : Binding Sites, Plasmids, Promoter Regions, Genetic, SARS Virus.
- metabolism : SARS Virus.
- methods : Mutagenesis, Site-Directed.
- chemical , physiology : Nucleocapsid Proteins.
- Amino Acid Sequence, Cell Line, Chromatin Immunoprecipitation, Electrophoretic Mobility Shift Assay, Gene Expression Regulation, Enzymologic, Humans, Mutation, Protein Binding, Transcriptional Activation.
Abstract
SARS-associated coronavirus (SARS-CoV) causes inflammation and damage to the lungs resulting in severe acute respiratory syndrome. To evaluate the molecular mechanisms behind this event, we investigated the roles of SARS-CoV proteins in regulation of the proinflammatory factor, cyclooxygenase-2 (COX-2). Individual viral proteins were tested for their abilities to regulate COX-2 gene expression. Results showed that the COX-2 promoter was activated by the nucleocapsid (N) protein in a concentration-dependent manner. Western blot analysis indicated that N protein was sufficient to stimulate the production of COX-2 protein in mammalian cells. COX-2 promoter mutations suggested that activation of COX-2 transcription depended on two regulatory elements, a nuclear factor-kappa B (NF-kappaB) binding site, and a CCAAT/enhancer binding protein (C/EBP) binding site. Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) demonstrated that SARS-CoV N protein bound directly to these regulatory sequences. Protein mutation analysis revealed that a Lys-rich motif of N protein acted as a nuclear localization signal and was essential for the activation of COX-2. In addition, a Leu-rich motif was found to be required for the N protein function. A sequence of 68 residuals was identified as a potential DNA-binding domain essential for activating COX-2 expression. We propose that SARS-CoV N protein causes inflammation of the lungs by activating COX-2 gene expression by binding directly to the promoter resulting in inflammation through multiple COX-2 signaling cascades.
DOI: 10.1016/j.biocel.2006.02.003
PubMed: 16546436
Affiliations:
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Le document en format XML
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<term>CCAAT-Enhancer-Binding Protein-beta (metabolism)</term>
<term>Cell Line</term>
<term>Chromatin Immunoprecipitation</term>
<term>Cyclooxygenase 2 (genetics)</term>
<term>Cyclooxygenase 2 (metabolism)</term>
<term>Electrophoretic Mobility Shift Assay</term>
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<term>Humans</term>
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<term>Nucleocapsid Proteins (metabolism)</term>
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<term>Protein Binding</term>
<term>SARS Virus (genetics)</term>
<term>SARS Virus (metabolism)</term>
<term>Transcriptional Activation</term>
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<term>Cyclooxygenase 2 (génétique)</term>
<term>Cyclooxygenase 2 (métabolisme)</term>
<term>Facteur de transcription NF-kappa B (métabolisme)</term>
<term>Humains</term>
<term>Immunoprécipitation de la chromatine</term>
<term>Liaison aux protéines</term>
<term>Lignée cellulaire</term>
<term>Mutagenèse dirigée ()</term>
<term>Mutation</term>
<term>Plasmides (génétique)</term>
<term>Protéine bêta de liaison aux séquences stimulatrices de type CCAAT (métabolisme)</term>
<term>Protéines nucléocapside (génétique)</term>
<term>Protéines nucléocapside (métabolisme)</term>
<term>Protéines nucléocapside (physiologie)</term>
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<term>Régulation de l'expression des gènes codant pour des enzymes</term>
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<term>Séquence d'acides aminés</term>
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<term>Virus du SRAS (métabolisme)</term>
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<term>Cyclooxygenase 2</term>
<term>NF-kappa B</term>
<term>Nucleocapsid Proteins</term>
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<term>Protéines nucléocapside</term>
<term>Régions promotrices (génétique)</term>
<term>Sites de fixation</term>
<term>Virus du SRAS</term>
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<term>Protéines nucléocapside</term>
<term>Virus du SRAS</term>
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<term>Lignée cellulaire</term>
<term>Mutagenèse dirigée</term>
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<term>Régulation de l'expression des gènes codant pour des enzymes</term>
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<front><div type="abstract" xml:lang="en">SARS-associated coronavirus (SARS-CoV) causes inflammation and damage to the lungs resulting in severe acute respiratory syndrome. To evaluate the molecular mechanisms behind this event, we investigated the roles of SARS-CoV proteins in regulation of the proinflammatory factor, cyclooxygenase-2 (COX-2). Individual viral proteins were tested for their abilities to regulate COX-2 gene expression. Results showed that the COX-2 promoter was activated by the nucleocapsid (N) protein in a concentration-dependent manner. Western blot analysis indicated that N protein was sufficient to stimulate the production of COX-2 protein in mammalian cells. COX-2 promoter mutations suggested that activation of COX-2 transcription depended on two regulatory elements, a nuclear factor-kappa B (NF-kappaB) binding site, and a CCAAT/enhancer binding protein (C/EBP) binding site. Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) demonstrated that SARS-CoV N protein bound directly to these regulatory sequences. Protein mutation analysis revealed that a Lys-rich motif of N protein acted as a nuclear localization signal and was essential for the activation of COX-2. In addition, a Leu-rich motif was found to be required for the N protein function. A sequence of 68 residuals was identified as a potential DNA-binding domain essential for activating COX-2 expression. We propose that SARS-CoV N protein causes inflammation of the lungs by activating COX-2 gene expression by binding directly to the promoter resulting in inflammation through multiple COX-2 signaling cascades.</div>
</front>
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<tree><noCountry><name sortKey="Hao, Qian" sort="Hao, Qian" uniqKey="Hao Q" first="Qian" last="Hao">Qian Hao</name>
<name sortKey="Mu, Yongxin" sort="Mu, Yongxin" uniqKey="Mu Y" first="Yongxin" last="Mu">Yongxin Mu</name>
<name sortKey="Timani, Khalid Amine" sort="Timani, Khalid Amine" uniqKey="Timani K" first="Khalid Amine" last="Timani">Khalid Amine Timani</name>
<name sortKey="Wu, Jianguo" sort="Wu, Jianguo" uniqKey="Wu J" first="Jianguo" last="Wu">Jianguo Wu</name>
<name sortKey="Ye, Linbai" sort="Ye, Linbai" uniqKey="Ye L" first="Linbai" last="Ye">Linbai Ye</name>
<name sortKey="Zhu, Ying" sort="Zhu, Ying" uniqKey="Zhu Y" first="Ying" last="Zhu">Ying Zhu</name>
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<country name="République populaire de Chine"><region name="Hubei"><name sortKey="Yan, Xiaohong" sort="Yan, Xiaohong" uniqKey="Yan X" first="Xiaohong" last="Yan">Xiaohong Yan</name>
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