Machitani Mitsuhiro, Sakurai Fuminori, Wakabayashi Keisaku, Tomita Kyoko, Tachibana Masashi, Mizuguchi Hiroyuki
Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.
Laboratory of Regulatory Sciences for Oligonucleotide Therapeutics, Clinical Drug Development Unit, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka, 565-0871, Japan.
Sci Rep. 2016 Jun 7;6:27598. doi: 10.1038/srep27598.
In various organisms, including nematodes and plants, RNA interference (RNAi) is a defense system against virus infection; however, it is unclear whether RNAi functions as an antivirus system in mammalian cells. Rather, a number of DNA viruses, including herpesviruses, utilize post-transcriptional silencing systems for their survival. Here we show that Dicer efficiently suppresses the replication of adenovirus (Ad) via cleavage of Ad-encoding small RNAs (VA-RNAs), which efficiently promote Ad replication via the inhibition of eIF2α phosphorylation, to viral microRNAs (mivaRNAs). The Dicer knockdown significantly increases the copy numbers of VA-RNAs, leading to the efficient inhibition of eIF2α phosphorylation and the subsequent promotion of Ad replication. Conversely, overexpression of Dicer significantly inhibits Ad replication. Transfection with mivaRNA does not affect eIF2α phosphorylation or Ad replication. These results indicate that Dicer-mediated processing of VA-RNAs leads to loss of activity of VA-RNAs for enhancement of Ad replication and that Dicer functions as a defence system against Ad in mammalian cells.
在包括线虫和植物在内的多种生物体中,RNA干扰(RNAi)是一种抵御病毒感染的防御系统;然而,RNAi在哺乳动物细胞中是否作为抗病毒系统发挥作用尚不清楚。相反,包括疱疹病毒在内的许多DNA病毒利用转录后沉默系统来生存。在此我们表明,Dicer通过将腺病毒(Ad)编码的小RNA(VA-RNAs)切割成病毒微小RNA(mivaRNAs),有效地抑制了腺病毒(Ad)的复制,而VA-RNAs通过抑制eIF2α磷酸化有效地促进了Ad复制。Dicer基因敲低显著增加了VA-RNAs的拷贝数,导致eIF2α磷酸化受到有效抑制,进而促进了Ad复制。相反,Dicer的过表达显著抑制了Ad复制。用mivaRNA转染不影响eIF2α磷酸化或Ad复制。这些结果表明,Dicer介导的VA-RNAs加工导致VA-RNAs丧失增强Ad复制的活性,并且Dicer在哺乳动物细胞中作为抵御Ad的防御系统发挥作用。