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

Xiaohong Yan, Qian Hao, Yongxin Mu, Khalid A. Timani, Linbai Ye, Ying Zhu, Jianguo Wu

Research output: Contribution to journalArticleResearchpeer-review

44 Citations (Scopus)

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-κB) 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.

Original languageEnglish
Pages (from-to)1417-1428
Number of pages12
JournalInternational Journal of Biochemistry and Cell Biology
Volume38
Issue number8
DOIs
StatePublished - 8 May 2006

Fingerprint

SARS Virus
CCAAT-Enhancer-Binding Proteins
Nucleocapsid Proteins
NF-kappa B
Cyclooxygenase 2
Proteins
Gene expression
Chemical activation
Binding Sites
Inflammation
Gene Expression
Severe Acute Respiratory Syndrome
Nucleocapsid
Nuclear Localization Signals
Electrophoretic mobility
Amino Acid Motifs
Mutation
Chromatin Immunoprecipitation
Electrophoretic Mobility Shift Assay
Viral Proteins

Keywords

  • Cyclooxygenase-2
  • Gene regulation
  • Inflammation
  • N protein
  • Pathogenesis
  • SARS-CoV
  • Virus infection

Cite this

@article{08c27a8c45dc4a90b26f64fe9e0eee94,
title = "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",
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-κB) 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.",
keywords = "Cyclooxygenase-2, Gene regulation, Inflammation, N protein, Pathogenesis, SARS-CoV, Virus infection",
author = "Xiaohong Yan and Qian Hao and Yongxin Mu and Timani, {Khalid A.} and Linbai Ye and Ying Zhu and Jianguo Wu",
year = "2006",
month = "5",
day = "8",
doi = "10.1016/j.biocel.2006.02.003",
language = "English",
volume = "38",
pages = "1417--1428",
journal = "International Journal of Biochemistry and Cell Biology",
issn = "1357-2725",
publisher = "Elsevier Ltd",
number = "8",

}

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. / Yan, Xiaohong; Hao, Qian; Mu, Yongxin; Timani, Khalid A.; Ye, Linbai; Zhu, Ying; Wu, Jianguo.

In: International Journal of Biochemistry and Cell Biology, Vol. 38, No. 8, 08.05.2006, p. 1417-1428.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - 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

AU - Yan, Xiaohong

AU - Hao, Qian

AU - Mu, Yongxin

AU - Timani, Khalid A.

AU - Ye, Linbai

AU - Zhu, Ying

AU - Wu, Jianguo

PY - 2006/5/8

Y1 - 2006/5/8

N2 - 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-κB) 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.

AB - 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-κB) 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.

KW - Cyclooxygenase-2

KW - Gene regulation

KW - Inflammation

KW - N protein

KW - Pathogenesis

KW - SARS-CoV

KW - Virus infection

UR - http://www.scopus.com/inward/record.url?scp=33646168070&partnerID=8YFLogxK

U2 - 10.1016/j.biocel.2006.02.003

DO - 10.1016/j.biocel.2006.02.003

M3 - Article

VL - 38

SP - 1417

EP - 1428

JO - International Journal of Biochemistry and Cell Biology

JF - International Journal of Biochemistry and Cell Biology

SN - 1357-2725

IS - 8

ER -