Long-Read Sequencing of Human Cytomegalovirus Transcriptome Reveals RNA Isoforms Carrying Distinct Coding Potentials.

Long-Read Sequencing of Human Cytomegalovirus Transcriptome Reveals RNA Isoforms Carrying Distinct Coding Potentials.
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DOI:
10.1038/s41598-017-16262-z
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发表时间:
2017-11-22
期刊:
影响因子:
4.6
通讯作者:
Boldogkői Z
Boldogkői Z
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Balázs Z;Tombácz D;Szűcs A;Csabai Z;Megyeri K;Petrov AN;Snyder M;Boldogkői Z

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人巨细胞病毒(HCMV)是一种普遍存在的人类致病疱疹病毒。完整的病毒基因组在感染期间转录活跃;然而,其转录组的很大一部分尚未被注释。在这项工作中,我们应用太平洋生物科学公司的扩增异构体测序技术来鉴定HCMV Towne vars株的裂解转录组。我们开发了一种使用长读测序数据进行转录注释的流水线。我们确定了248个转录起始点、116个转录终止点和80个剪接事件。利用这些信息,我们已经注释了291个先前未描述或仅部分注释的转录本异构体,包括8个新的反义转录本及其异构体,以及短重复区域中的一个新转录本(RS2),部分反义RS1。与其他生物类似,我们在HCMV中发现了高度的转录多样性,许多转录本之间只有轻微的差异。将我们的转录组图谱结果与早期的核糖体足迹分析进行比较,我们得出的结论是,大多数转录本包含多个翻译活性ORF,并且大多数异构体包含独特的ORF组合。基于这些结果,我们认为这种转录多样性的一个重要功能可能是在翻译水平上提供一种调控机制。
The human cytomegalovirus (HCMV) is a ubiquitous, human pathogenic herpesvirus. The complete viral genome is transcriptionally active during infection; however, a large part of its transcriptome has yet to be annotated. In this work, we applied the amplified isoform sequencing technique from Pacific Biosciences to characterize the lytic transcriptome of HCMV strain Towne varS. We developed a pipeline for transcript annotation using long-read sequencing data. We identified 248 transcriptional start sites, 116 transcriptional termination sites and 80 splicing events. Using this information, we have annotated 291 previously undescribed or only partially annotated transcript isoforms, including eight novel antisense transcripts and their isoforms, as well as a novel transcript (RS2) in the short repeat region, partially antisense to RS1. Similarly to other organisms, we discovered a high transcriptional diversity in HCMV, with many transcripts only slightly differing from one another. Comparing our transcriptome profiling results to an earlier ribosome footprint analysis, we have concluded that the majority of the transcripts contain multiple translationally active ORFs, and also that most isoforms contain unique combinations of ORFs. Based on these results, we propose that one important function of this transcriptional diversity may be to provide a regulatory mechanism at the level of translation.
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