Abstract

Novel duck reovirus (NDRV) causes severe economic losses to the duck industry, which is characterized by hemorrhagic spots and necrotic foci of the livers and spleens. DEAD-box helicase 1 (DDX1) plays a critical role in the innate immune system against viral infection. However, the role of duck DDX1 (duDDX1) in anti-RNA virus infection, especially in the anti-NDRV infection, has yet to be elucidated. In the present study, the full-length cDNA of duDDX1 (2223 bp encode 740 amino acids) was firstly cloned from the spleen of healthy Cherry valley ducks, and the phylogenetic tree indicated that the duDDX1 has the closest relationship with Anas platyrhynchos in the bird branch. The duDDX1 mRNA was widely distributed in all tested tissues, especially in the duodenum, liver, and spleen. Overexpression of duDDX1 in primary duck embryo fibroblast (DEF) cells triggered the activation of transcription factors IRF-7 and NF-κB, as well as IFN-β expression, and the expression of the Toll-like receptors (TLR2, TLR3, and TLR4) was significantly increased. Importantly, after overexpressing or knocking down duDDX1 and infecting NDRV in DEF cells, duDDX1 inhibits the replication of NDRV virus and also regulates the expression of pattern recognition receptors and cytokines. This indicates that duDDX1 may play an important role in the innate immune response of ducks to NDRV. Collectively, we first cloned DDX1 from ducks and analyzed its biological functions. Secondly, we proved that duck DDX1 participates in anti-NDRV infection, and innovated new ideas for the prevention and control of duck virus infection.

Highlights

  • The new duck reovirus (NDRV) is a double-stranded RNA virus and a member of the Orthoreovirus genus in the Reoviridae family

  • DuDDX1 is involved in innate immunity To determine the role of duck DDX1 (duDDX1) in duck innate immunity response, the pcDNA3.0-duDDX1-Flag or empty vector were transfected into duck embryo fibroblast (DEF) cells, and DEF cells were collected at designated time points for subsequent experiments (Figure 5)

  • DuDDX1 encoded an amino-acid sequence of 740 residues

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Summary

Introduction

The new duck reovirus (NDRV) is a double-stranded RNA (dsRNA) virus and a member of the Orthoreovirus genus in the Reoviridae family. The genome of the virus consists of 10 genome segments, L1-3, M1-3 and S1-4 [1]. The encoded NDRV S1 segment is similar to avian reovirus (ARV) [2], with three partially overlapping open reading frames (ORFs), encoding p10, p18, and σC, In the process of virus replication, first, the virusencoded dsRNA-dependent RNA polymerase uses the negative-strand RNA of the viral genome as a template to synthesize mRNA. Viral mRNA plays a dual function in the infected cell, synthesizing viral proteins in the ribosome and as a template to synthesize viral genome negativestrand RNA. NDRV infection mainly causes hemorrhage and necrosis of the liver and spleen, and may lead to the transfer of intestinal flora1 [5], which is different from diseases

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