Multi-platform analysis reveals a complex transcriptome architecture of a circovirus

Virus Res. 2017 Jun 2:237:37-46. doi: 10.1016/j.virusres.2017.05.010. Epub 2017 May 24.

Abstract

In this study, we used Pacific Biosciences RS II long-read and Illumina HiScanSQ short-read sequencing technologies for the characterization of porcine circovirus type 1 (PCV-1) transcripts. Our aim was to identify novel RNA molecules and transcript isoforms, as well as to determine the exact 5'- and 3'-end sequences of previously described transcripts with single base-pair accuracy. We discovered a novel 3'-UTR length isoform of the Cap transcript, and a non-spliced Cap transcript variant. Additionally, our analysis has revealed a 3'-UTR isoform of Rep and two 5'-UTR isoforms of Rep' transcripts, and a novel splice variant of the longer Rep' transcript. We also explored two novel long transcripts, one with a previously identified splice site, and a formerly undetected mRNA of ORF3. Altogether, our methods have identified nine novel RNA molecules, doubling the size of PCV-1 transcriptome that had been known before. Additionally, our investigations revealed an intricate pattern of transcript overlapping, which might produce transcriptional interference between the transcriptional machineries of adjacent genes, and thereby may potentially play a role in the regulation of gene expression in circoviruses.

Keywords: Isoform sequencing; Long-read sequencing; PacBio sequencing; Porcine circovirus; Short-read sequencing; Transcriptional interference; Transcriptome.

MeSH terms

  • Animals
  • Cell Line
  • Circovirus / physiology*
  • Gene Expression Profiling*
  • RNA, Viral / analysis*
  • RNA, Viral / genetics
  • Sequence Analysis, RNA
  • Swine
  • Transcription, Genetic*
  • Virus Replication*

Substances

  • RNA, Viral

Supplementary concepts

  • Porcine circovirus 1