Abstract
BackgroundToll-like receptors (TLRs) play an important role in detecting pathogen-associated molecular patterns (PAMPs). Among the TLRs, TLR3 is involved in the recognition of double-stranded RNA. This study was designed to explore the relationship between duTLR3 and duck reovirus (DRV) infection.MethodsIn this study, we cloned and performed a molecular characterization of the complete sequence of Peking duck TLR3 (duTLR3). The expression level of duTLR3 was also determined, along with the relative levels of Mx and IFN-α mRNA after DRV infection.ResultsThe duTLR3 gene is 2776-bp long and encodes an 895-amino-acid-long protein. Sequence analysis of the product revealed the complete transcript of Peking duck TLR3, including the 88-bp 5′UTR, the 2688-bp coding sequence (ORF), and the 76-bp 3′UTR and poly(A) tail. DuTLR3 was found to share a high amino acid sequence similarity with TLR3 from Jing ding duck (99.6 %), Muscovy duck (97.1 %) and chicken (86.3 %). Additionally, the tissue distribution of duTLR3 suggested that it was abundantly expressed in various tissues, especially in the trachea, esophagus and pancreatic gland. Duck reovirus (DRV) infection resulted in high mRNA expression levels of duTLR3 in the spleen, liver, lung and brain.ConclusionThese results suggest that duTLR3 may play an important role in anti-viral defense mechanisms.Electronic supplementary materialThe online version of this article (doi:10.1186/s12985-015-0434-x) contains supplementary material, which is available to authorized users.
Highlights
Toll-like receptors (TLRs) play an important role in detecting pathogen-associated molecular patterns (PAMPs)
The full-length duck TLR3 (duTLR3) cDNA sequence was obtained from Peking duck by homologous cloning and Rapid amplification of cDNA ends (RACE) techniques
Sequence analysis of the product revealed the complete transcript of Peking duck duTLR3, including the 88-bp 5′UTR, the 2688-bp coding sequence (ORF), and the 76-bp 3′UTR and poly(A) tail
Summary
Toll-like receptors (TLRs) play an important role in detecting pathogen-associated molecular patterns (PAMPs). Among the TLRs, TLR3 is involved in the recognition of double-stranded RNA. The initial sensing of infection is mediated by innate pattern recognition receptors (PRRs), which include Toll-like receptors (TLRs), RIG-I-like receptors (RLRs), NOD-like receptors and C-type lectin receptors. TLRs play an important role in detecting pathogen-associated molecular patterns (PAMPs). Among the TLRs, TLR3 is involved in the recognition of double-stranded RNA, which is a molecular pattern produced by many viruses and can be considered a viral PAMP [1]. The interaction of dsRNA with the TLR3-ECD leads to receptor dimerization and recruitment of the adapter molecule TRIF to the cytoplasmic domain of TLR3, which is known as a TIR (Toll/ interleukin-1 receptor) domain due to its homology with the signaling domains of the IL-1 receptor and plant resistance proteins [2]. TRIF initiates signaling pathways that activate the downstream transcription factors IRF3, AP-1 and NF-κB, which in turn trigger the expression and secretion of type I interferons, inflammatory cytokines and chemokines [3]
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