James Hogg Research Center, Providence Heart and Lung Institute, St. Paul's Hospital, University of British Columbia, Vancouver, British Columbia, Canada.
FASEB J. 2013 Jul;27(7):2777-87. doi: 10.1096/fj.12-226498. Epub 2013 Apr 9.
Coxsackievirus B3 (CVB3) is a causative agent of viral myocarditis, hepatitis, pancreatitis, and meningitis in humans. The adenosine-uridine (AU)-rich element RNA binding factor 1 (AUF1) is an integral component in the regulation of gene expression. AUF1 destabilizes mRNAs and targets them for degradation by binding to AU-rich elements in the 3' untranslated region (UTR) of mRNAs. The 3'-UTR of the CVB3 genome contains canonical AU-rich sequences, raising the possibility that CVB3 RNA may also be subjected to AUF1-mediated degradation. Here, we reported that CVB3 infection led to cytoplasmic redistribution and cleavage of AUF1. These events are independent of CVB3-induced caspase activation but require viral protein production. Overexpression of viral protease 2A reproduced CVB3-induced cytoplasmic redistribution of AUF1, while in vitro cleavage assay revealed that viral protease 3C contributed to AUF1 cleavage. Furthermore, we showed that knockdown of AUF1 facilitated viral RNA, protein, and progeny production, suggesting an antiviral property for AUF1 against CVB3 infection. Finally, an immunoprecipitation study demonstrated the physical interaction between AUF1 and the 3'-UTR of CVB3, potentially targeting CVB3 genome toward degradation. Together, our results suggest that cleavage of AUF1 may be a strategy employed by CVB3 to enhance the stability of its viral genome.
柯萨奇病毒 B3(CVB3)是人类病毒性心肌炎、肝炎、胰腺炎和脑膜炎的病原体。腺嘌呤-尿嘧啶(AU)丰富元件 RNA 结合因子 1(AUF1)是调节基因表达的一个组成部分。AUF1 通过结合 mRNA 3'非翻译区(UTR)中的 AU 丰富元件,使 mRNA 不稳定,并将其靶向降解。CVB3 基因组的 3'-UTR 含有典型的 AU 丰富序列,这使得 CVB3 RNA 也可能受到 AUF1 介导的降解。在这里,我们报道了 CVB3 感染导致 AUF1 的细胞质重分布和切割。这些事件不依赖于 CVB3 诱导的半胱天冬酶激活,但需要病毒蛋白的产生。病毒蛋白酶 2A 的过表达复制了 CVB3 诱导的 AUF1 细胞质重分布,而体外切割实验表明病毒蛋白酶 3C 有助于 AUF1 的切割。此外,我们还表明,AUF1 的敲低促进了病毒 RNA、蛋白和后代的产生,表明 AUF1 对 CVB3 感染具有抗病毒特性。最后,一项免疫沉淀研究表明 AUF1 与 CVB3 的 3'-UTR 之间存在物理相互作用,可能将 CVB3 基因组靶向降解。总之,我们的研究结果表明,AUF1 的切割可能是 CVB3 用来增强其病毒基因组稳定性的一种策略。