Lu Ying, Ma Yanping, Liu Zhongyang, Han Liying, Gao Shuang, Zheng Bo, Liu Chang, Qi Ying, Sun Zhengrong, Huang Yujing, Ruan Qiang
Virus Laboratory, Affiliated Shengjing Hospital, China Medical University, Shenyang, Liaoning, 121000, China.
Department of Pathogen Biology, College of Basic Medical Sciences, Liaoning Medical University, Jinzhou, Liaoning, 121000, China.
Virus Genes. 2016 Jun;52(3):334-45. doi: 10.1007/s11262-016-1308-z. Epub 2016 Mar 1.
Among all the human cytomegalovirus (HCMV) gene families, US12 family is relatively undefined in their transcriptional profile and biological functions. In this study, the transcription pattern and characteristics of HCMV US12-US17 gene region were studied extensively. Twenty-three clones harboring US12 cDNA sequence were screened out from a late cDNA library of an HCMV clinical isolate, Han. Using a set of US12-US17 gene-specific probes, six transcripts from US12-US17 locus were detected by northern blot at late kinetics of the clinical isolate. One additional transcript was found in late RNA of HCMV strain AD169. No evidence showing these transcripts contain introns by reverse transcription PCR. 3' and 5' termini of these transcripts were confirmed by Rapid Amplification of cDNA Ends. A novel protein-coding region was predicted in the shorter US14 transcript with an alternative in-frame 5' translation initiation site compared to that of the previously predicted US14 ORF. Our findings demonstrate the existence of a cluster of 3' coterminal unspliced transcripts with distinct 5' transcriptional initiation sites originated from US12-US17 gene region in the late infection phase of an HCMV clinical strain.
在所有人类巨细胞病毒(HCMV)基因家族中,US12家族的转录谱和生物学功能相对不明确。在本研究中,对HCMV US12-US17基因区域的转录模式和特征进行了广泛研究。从一株HCMV临床分离株Han的晚期cDNA文库中筛选出23个含有US12 cDNA序列的克隆。使用一组US12-US17基因特异性探针,通过Northern印迹法在该临床分离株的晚期动力学阶段检测到来自US12-US17位点的6种转录本。在HCMV AD169株的晚期RNA中发现了另外一种转录本。通过逆转录PCR没有证据表明这些转录本含有内含子。通过cDNA末端快速扩增法确定了这些转录本的3'和5'末端。与先前预测的US14开放阅读框相比,在较短的US14转录本中预测到一个新的蛋白质编码区域,其5'翻译起始位点存在框内替代。我们的研究结果表明,在HCMV临床株感染后期,存在一组源自US12-US17基因区域的具有不同5'转录起始位点的3'共末端未剪接转录本。