Torma Gábor, Tombácz Dóra, Csabai Zsolt, Göbhardter Dániel, Deim Zoltán, Snyder Michael, Boldogkői Zsolt
Department of Medical Biology, Faculty of Medicine, University of Szeged, 6720 Szeged, Hungary.
Department of Genetics, School of Medicine, Stanford University, Stanford, CA 94304, USA.
Pathogens. 2021 Feb 20;10(2):242. doi: 10.3390/pathogens10020242.
In the last couple of years, the implementation of long-read sequencing (LRS) technologies for transcriptome profiling has uncovered an extreme complexity of viral gene expression. In this study, we carried out a systematic analysis on the pseudorabies virus transcriptome by combining our current data obtained by using Pacific Biosciences Sequel and Oxford Nanopore Technologies MinION sequencing with our earlier data generated by other LRS and short-read sequencing techniques. As a result, we identified a number of novel genes, transcripts, and transcript isoforms, including splice and length variants, and also confirmed earlier annotated RNA molecules. One of the major findings of this study is the discovery of a large number of 5'-truncations of larger putative mRNAs being 3'-co-terminal with canonical mRNAs of PRV. A large fraction of these putative RNAs contain in-frame ATGs, which might initiate translation of N-terminally truncated polypeptides. Our analyses indicate that CTO-S, a replication origin-associated RNA molecule is expressed at an extremely high level. This study demonstrates that the PRV transcriptome is much more complex than previously appreciated.
在过去几年中，用于转录组分析的长读长测序（LRS）技术的应用揭示了病毒基因表达的极端复杂性。在本研究中，我们通过将使用太平洋生物科学公司的Sequel和牛津纳米孔技术公司的MinION测序获得的当前数据与我们早期通过其他LRS和短读长测序技术生成的数据相结合，对伪狂犬病病毒转录组进行了系统分析。结果，我们鉴定出了许多新基因、转录本和转录异构体，包括剪接变体和长度变体，同时也证实了先前注释的RNA分子。本研究的一个主要发现是发现了大量较大的假定mRNA的5'端截短，这些截短与伪狂犬病病毒的经典mRNA在3'端共末端。这些假定RNA中的很大一部分含有符合读框的ATG，这可能启动N端截短多肽的翻译。我们的分析表明，复制起始点相关RNA分子CTO-S的表达水平极高。本研究表明，伪狂犬病病毒转录组比以前认为的要复杂得多。
