Identification and characterization of a novel group of natural antisense transcripts from RNA1.2 gene locus of human cytomegalovirus
Identification and characterization of a novel group of natural antisense transcripts from RNA1.2 gene locus of human cytomegalovirus
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人巨细胞病毒 RNA1.2 基因座中一组新的天然反义转录物的鉴定和表征
DOI：
10.1097/cm9.0000000000000299
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发表时间：
2019
影响因子：
6.1
通讯作者：
Ruan Qiang
中科院分区：
文献类型：
--
作者：
Liu Bing Yang;Ma Yan Ping;Qi Ying;Huang Yu Jing;Liu Zhong Yang;Lu Meng Qing;Ruan Qiang
背景天然反义转录本（NAT）是从注释基因的互补DNA链转录而来，发挥基因表达的调节功能。越来越多的研究认识到反义转录广泛存在于人类巨细胞病毒 (HCMV) 基因组中，而 RNA1.2 基因座的反义转录从未被研究过。在本研究中，在临床分离的 HCMV 毒株中研究了 RNA1.2 反义链的转录。方法进行链特异性高通量 RNA 测序 (RNA-seq) 以寻找可能的反义转录本 (AST)。为了分析和可视化 RNA-seq 数据集，应用了 Integrative Genomics Viewer 软件。为了证实这些可能性，使用了Northern印迹和cDNA末端快速扩增(RACE)。结果使用从感染HCMV临床分离株HAN的人胚胎肺成纤维细胞中提取的RNA，通过RNA测序检测到RNA1.2基因座相反链的转录。通过 Northern 印迹和 RACE 分析对至少三种 HCMV NAT 进行了表征，分别称为 RNA1.2 AST 1、RNA1.2 AST2 和 RNA1.2 AST3。这些 RNA1.2 AST 在 HCMV 感染后期从 RNA1.2 基因座的互补链定向。这些转录物的5'-和3'-末端位于预测的RNA1.2基因的相反序列内。结论一簇新的NAT是从HCMV RNA1.2基因区域的相反序列转录而来的。
BackgroundNatural anti-sense transcripts (NATs), which are transcribed from the complementary DNA strand of annotated genes, exert regulatory function of gene expression. Increasing studies recognized anti-sense transcription widespread throughout human cytomegalovirus (HCMV) genome, whereas the anti-sense transcription of RNA1.2 gene locus has never been investigated. In this study, the transcription of the RNA1.2 anti-sense strand was investigated in clinically isolated HCMV strain.MethodsStrand-specific high-through RNA-sequencing (RNA-seq) was performed to find possible anti-sense transcripts (ASTs). For analyzing and visualization of RNA-seq data sets, Integrative Genomics Viewer software was applied. To confirm these possibilities, Northern blotting and rapid amplification of cDNA ends (RACE) were used.ResultsTranscription of the opposite strand of RNA1.2 gene locus was detected by RNA-sequencing using RNAs extracted from human embryonic lung fibroblasts infected with HCMV clinical isolate HAN. At least three HCMV NATs, named RNA1.2 AST 1, RNA1.2 AST2, and RNA1.2 AST3, were characterized by Northern blotting and RACE analyses. These RNA1.2 ASTs orientated from the complementary strand of RNA1.2 locus during the late phase of HCMV infection. The 5′- and 3′-termini of these transcripts were located within the opposite sequence of the predicted RNA1.2 gene.ConclusionA cluster of novel NATs was transcribed from the opposite sequence of the HCMV RNA1.2 gene region.