Molecular cloning and functional characterization of duck DDX41
Molecular cloning and functional characterization of duck DDX41
复制标题
鸭DDX41的分子克隆及功能表征
DOI：
10.1016/j.dci.2018.07.015
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发表时间：
2018-11-01
影响因子：
2.9
通讯作者：
Jin, Hui
中科院分区：
文献类型：
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作者：
Li, Yaqian;Li, Huilin;Jin, Hui
DEAD(Asp-Glu-Ala-Asp)盒多肽41(DDX41)是DExD/H-box解旋酶家族的一种受体，可作为细胞内DNA感受器，诱导哺乳动物和鱼类产生I型干扰素。然而，禽类DDX41在先天免疫应答中的作用尚不清楚。本研究首次克隆了鸭DDX41(DuDDX41)的全长cDNA序列，编码618个氨基酸残基的蛋白质，与斑雀和鸡的DDX41S具有较高的序列相似性。DuDDX41mRNA广泛分布于所有受试组织，尤其是大脑、小脑和肝脏。DuDDX41的过表达触发了转录因子IRF1和NF-kappa B的激活，以及干扰素-β在Defs中的表达。DuDDX41的DEADc结构域在鸭I型干扰素信号通路中发挥着极其重要的作用。通过siRNA沉默的方法敲除duDDX41可显著降低由聚(da：dt)或鸭肠炎病毒(DEV)刺激的干扰素-β的表达。此外，DEV在duDDX41表达的Defs中的复制被显著抑制，而在DDX41基因敲除的Defs中被增强。这些结果表明，DDX41是一种重要的胞质DNA感受器，在鸭抗病毒天然免疫反应中发挥重要作用。
DEAD (Asp-Glu-Ala-Asp) box polypeptide 41 (DDX41), a receptor belonging to DExD/H-box helicase family, acts as an intracellular DNA sensor and induces type I IFN production in mammals and fish. However, the function of avian DDX41 in innate immune response is still unknown. In this study, the full-length duck DDX41 (duDDX41) cDNA sequence was cloned for the first time and encoded a putative protein of 618 amino acid residues which showed the high sequence similarity with both zebra finch and chicken DDX41s. The duDDX41 mRNA was widely distributed in all tested tissues, especially the cerebrum, cerebellum, and liver. Overexpression of duDDX41 triggered the activation of transcription factors IRF1 and NF-kappa B, as well as IFN-beta expression in DEFs. The DEADc domain of duDDX41 played an extremely vital role in duck type I IFN signaling pathway. Knockdown of duDDX41 by siRNA silencing dramatically decreased IFN-beta expression stimulated by poly(dA:dT) or duck enteritis virus (DEV). In addition, the replication of DEV was significantly inhibited in duDDX41-expressed DEFs and was enhanced in DDX41 knockdown DEFs. These results suggest that DDX41 is an important cytosolic DNA sensor and plays a crucial role in duck antiviral innate immune response.