Mechanisms and roles by which IRF-3 mediates the regulation of ORMDL3 transcription in respiratory syncytial virus infection. (June 2017)

• Record Type: Journal Article
• Authors: Wang, Xiao-Hua; Shu, Jin; Jiang, Chun-Ming; Zhuang, Li-Li; Yang, Wei-Xia; Zhang, Hui-Weng; Wang, Lu-Lu; Li, Lin; Chen, Xiao-Qing; Jin, Rui; Zhou, Guo-Ping
• Is Part Of: International journal of biochemistry & cell biology. Volume 87(2017)
• Page Start: 8
• Page End: 17
• Publication Date: 2017-06
• Subjects: RSV respiratory syncytial virus -- ORMDL3 orosomucoid 1-like protein 3 -- IRF-3 interferon-regulatory factor-3 -- EMSA electrophoretic mobility shift assay -- ChIP chromatin immunoprecipitation -- NF-kB nuclear factor kB -- GWAS genome-wide association studies -- EBV Epstein–Barr virus -- HRV human rhinovirus -- PolyI:C polyinosinic-polycytidylic acid -- dsRNA double-stranded RNA -- DMEM Dulbecco's Modified Eagle Medium -- RPMI-1640 Roswell Park Memorial Institute Medium 1640 -- FBS foetal bovine serum -- CPE cytopathic effect -- PFU plaque forming unit -- MOI multiplicities of infection -- siRNA small interfering RNA -- Q-PCR quantitative real-time PCR -- BSA bovine serum albumin -- PBS phosphate-buffered saline -- ER endoplasmic reticulum; Orosomucoid 1-like protein 3 (ORMDL3) -- Respiratory syncytial virus (RSV) -- Interferon-regulatory factor-3 (IRF-3) -- Transcriptional regulation -- Bronchiolitis; Biochemistry -- Periodicals; Cytology -- Periodicals; Biochemistry -- Periodicals; Cell Biology -- Periodicals; Biochimie -- Périodiques; Cytologie -- Périodiques; Biochimie; Cytologie; Biochemistry; Cytology; Ressource Internet (Descripteur de forme); Périodique électronique (Descripteur de forme); Periodicals; 572.05
• DOI: 10.1016/j.biocel.2017.03.007
• Languages: English
• ISSNs: 1357-2725

Abstract

Graphical abstract: Highlights: ORMDL3 expression was significantly increased in infants with RSV-induced bronchiolitis compared with uninfected controls. The proximal interferon-regulatory factor-3 (IRF-3) binding site in the promoter region of ORMDL3 positively regulated its transcription following exposure to RSV. IRF-3 mediated the upregulation of RSV-induced ORMDL3 expression. IRF-3 bound directly to the promoter of the ORMDL3 gene during RSV infection. Abstract: Respiratory syncytial virus (RSV) is the leading cause of bronchiolitis in infancy, which is a major risk factor for recurrent wheezing and asthma. Orosomucoid 1-like protein 3 ( ORMDL3 ) has been reported to associate with virus-triggered recurrent wheezing and asthma in children. However, little is known about how ORMDL3 is involved into RSV infection. In this study, we showed that the mRNA expression of ORMDL3 is significantly increased in the peripheral blood lymphocytes of infants with RSV-induced bronchiolitis compared with uninfected controls, also increased in bronchial epithelial cells and lung fibroblasts following RSV infection in vitro . To investigate the underlying mechanisms of RSV-induced ORMDL3 expression, we performed in silico analysis of the binding sites of several transcription factors in the ORMDL3 promoter. The proximal interferon-regulatory factor-3 (IRF-3) binding site positively regulated ORMDL3 transcription following exposure to RSV, as determined through mutational analysis. Overexpression and RNA interference experiments targeting IRF-3 showed that it regulates the expression of ORMDL3 following RSV exposure. Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) assay showed that IRF-3 binds directly to the promoter of the ORMDL3 gene. Furthermore, we confirmed that expression of IRF-3 is significantly increased and shows a strong linear correlation with increased ORMDL3 in the peripheral blood lymphocytes from infants with RSV-induced bronchiolitis. Our results indicate that IRF-3 is an important regulator of ORMDL3 induction following RSV infection by binding directly to the promoter of ORMDL3, which may be implicated in the inflammatory and immune reactions involved in bronchiolitis and wheezing diseases.