Division of Biological Sciences, University of Montana, Missoula, MT 59812, USA.
Department of Chemistry and Biochemistry, University of Montana, Missoula, MT 59812, USA.
Viruses. 2021 Feb 26;13(3):367. doi: 10.3390/v13030367.
In recent years, transcriptome profiling studies have identified changes in host splicing patterns caused by viral invasion, yet the functional consequences of the vast majority of these splicing events remain uncharacterized. We recently showed that the host splicing landscape changes during Rift Valley fever virus MP-12 strain (RVFV MP-12) infection of mammalian cells. Of particular interest, we observed that the host mRNA for Rio Kinase 3 (RIOK3) was alternatively spliced during infection. This kinase has been shown to be involved in pattern recognition receptor (PRR) signaling mediated by RIG-I like receptors to produce type-I interferon. Here, we characterize RIOK3 as an important component of the interferon signaling pathway during RVFV infection and demonstrate that RIOK3 mRNA expression is skewed shortly after infection to produce alternatively spliced variants that encode premature termination codons. This splicing event plays a critical role in regulation of the antiviral response. Interestingly, infection with other RNA viruses and transfection with nucleic acid-based RIG-I agonists also stimulated RIOK3 alternative splicing. Finally, we show that specifically stimulating alternative splicing of the RIOK3 transcript using a morpholino oligonucleotide reduced interferon expression. Collectively, these results indicate that RIOK3 is an important component of the mammalian interferon signaling cascade and its splicing is a potent regulatory mechanism capable of fine-tuning the host interferon response.
近年来,转录组谱分析研究已经确定了病毒入侵引起的宿主剪接模式变化,但这些剪接事件中的绝大多数的功能后果仍未被描述。我们最近表明,在裂谷热病毒 MP-12 株(RVFV MP-12)感染哺乳动物细胞期间,宿主剪接景观发生变化。特别值得注意的是,我们观察到 Rio Kinase 3(RIOK3)的宿主 mRNA 在感染期间发生了可变剪接。已经表明该激酶参与了由 RIG-I 样受体介导的模式识别受体(PRR)信号传导,以产生 I 型干扰素。在这里,我们将 RIOK3 描述为 RVFV 感染期间干扰素信号通路的重要组成部分,并证明 RIOK3 mRNA 表达在感染后不久就会发生偏倚,产生编码提前终止密码子的可变剪接变体。这种剪接事件在调节抗病毒反应中起着关键作用。有趣的是,感染其他 RNA 病毒和用基于核酸的 RIG-I 激动剂转染也刺激了 RIOK3 的可变剪接。最后,我们表明,使用修饰的寡核苷酸特异性地刺激 RIOK3 转录本的可变剪接会降低干扰素的表达。总之,这些结果表明 RIOK3 是哺乳动物干扰素信号级联的重要组成部分,其剪接是一种能够微调宿主干扰素反应的强大调节机制。