From the The Wistar Institute, Philadelphia, Pennsylvania 19104; Institute for Genetic Medicine, Hokkaido University, Sapporo 060-0815, Japan.
From the The Wistar Institute, Philadelphia, Pennsylvania 19104.
J Biol Chem. 2010 Oct 22;285(43):33358-33370. doi: 10.1074/jbc.M110.138362. Epub 2010 Aug 17.
Certain primary transcripts of miRNA (pri-microRNAs) undergo RNA editing that converts adenosine to inosine. The Epstein-Barr virus (EBV) genome encodes multiple microRNA genes of its own. Here we report that primary transcripts of ebv-miR-BART6 (pri-miR-BART6) are edited in latently EBV-infected cells. Editing of wild-type pri-miR-BART6 RNAs dramatically reduced loading of miR-BART6-5p RNAs onto the microRNA-induced silencing complex. Editing of a mutation-containing pri-miR-BART6 found in Daudi Burkitt lymphoma and nasopharyngeal carcinoma C666-1 cell lines suppressed processing of miR-BART6 RNAs. Most importantly, miR-BART6-5p RNAs silence Dicer through multiple target sites located in the 3'-UTR of Dicer mRNA. The significance of miR-BART6 was further investigated in cells in various stages of latency. We found that miR-BART6-5p RNAs suppress the EBNA2 viral oncogene required for transition from immunologically less responsive type I and type II latency to the more immunoreactive type III latency as well as Zta and Rta viral proteins essential for lytic replication, revealing the regulatory function of miR-BART6 in EBV infection and latency. Mutation and A-to-I editing appear to be adaptive mechanisms that antagonize miR-BART6 activities.
某些 miRNA(pri-microRNAs)的初级转录本经历 RNA 编辑,将腺嘌呤转换为肌苷。 Epstein-Barr 病毒 (EBV) 基因组自身编码多个 microRNA 基因。在这里,我们报告 EBV 微小 RNA-BART6(pri-miR-BART6)的初级转录本在潜伏感染 EBV 的细胞中进行编辑。野生型 pri-miR-BART6 RNA 的编辑显著降低了 miR-BART6-5p RNA 加载到 microRNA 诱导的沉默复合物上的效率。在 Daudi Burkitt 淋巴瘤和鼻咽癌细胞系 C666-1 中发现的突变型 pri-miR-BART6 的编辑抑制了 miR-BART6 RNA 的加工。最重要的是,miR-BART6-5p RNA 通过位于 Dicer mRNA 3'-UTR 中的多个靶位点沉默 Dicer。我们在不同潜伏阶段的细胞中进一步研究了 miR-BART6 的意义。我们发现 miR-BART6-5p RNA 抑制了 EBNA2 病毒癌基因,该基因对于从免疫反应性较低的 I 型和 II 型潜伏状态向更具免疫反应性的 III 型潜伏状态以及裂解复制所必需的 Zta 和 Rta 病毒蛋白的转变是必需的,揭示了 miR-BART6 在 EBV 感染和潜伏中的调节功能。突变和 A-to-I 编辑似乎是拮抗 miR-BART6 活性的适应机制。