Fu Qiang, Shi Huijun, Chen Chuangfu
College of Veterinary Medicine, Xinjiang Agricultural University, Ürümqi, 830052, Xinjiang, China.
College of Animal Science and Technology, Shihezi University, Shihezi, 832003, Xinjiang, China.
Arch Virol. 2017 Feb;162(2):401-408. doi: 10.1007/s00705-016-3107-1. Epub 2016 Oct 20.
MicroRNAs (miRNAs) are an important class of small, non-coding RNAs that control target genes expression by degradation of target mRNAs or by inhibiting protein translation in many biological processes and cellular pathways. In a previous study, we found that miR-29b interfered with bovine viral diarrhea virus (BVDV) replication. However, the mechanisms of regulation of miR-29b expression are not well known. DNA methylation is an important epigenetic mechanism for silencing gene transcription, and plays an important role in promoter choice, protein expression, and regulation of miRNAs expression. In this study, we focused on the roles of DNA methylation of miR-29b promoter in regulating miR-29b expression and investigated the effects of DNA (cytosine-5) methyltransferase 1 (DNMT1) knockdown on miR-29b expression and BVDV (strain NADL) replication. Our results showed that methylation levels of miR-29b promoter were significantly decreased in BVDV NADL-infected MDBK cells. Furthermore, DNMT1 silencing significantly decreased the methylation levels of miR-29b promoter, up-regulated miR-29b expression and inhibited BVDV NADL replication, which supports the important roles of DNA methylation in regulating miRNA expression and further proves an evidence for our previous views.
微小RNA(miRNA)是一类重要的小型非编码RNA,在许多生物过程和细胞通路中,通过降解靶标mRNA或抑制蛋白质翻译来控制靶标基因的表达。在先前的一项研究中,我们发现miR-29b干扰牛病毒性腹泻病毒(BVDV)的复制。然而,miR-29b表达的调控机制尚不清楚。DNA甲基化是一种使基因转录沉默的重要表观遗传机制,在启动子选择、蛋白质表达和miRNA表达调控中发挥重要作用。在本研究中,我们聚焦于miR-29b启动子的DNA甲基化在调控miR-29b表达中的作用,并研究了DNA(胞嘧啶-5)甲基转移酶1(DNMT1)敲低对miR-29b表达和BVDV(NADL株)复制的影响。我们的结果表明,在BVDV NADL感染的MDBK细胞中,miR-29b启动子的甲基化水平显著降低。此外,DNMT1沉默显著降低了miR-29b启动子的甲基化水平,上调了miR-29b的表达并抑制了BVDV NADL的复制,这支持了DNA甲基化在调控miRNA表达中的重要作用,并进一步证明了我们先前观点的证据。