Abstract

Identification of the transcription start sites (TSSs) of a virus is of great importance to understand and dissect the mechanism of viral genome transcription but this often requires costly and laborious experiments. Many segmented negative-sense RNA viruses (sNSVs) cleave capped leader sequences from a large variety of mRNAs and use these cleaved leaders as primers for transcription in a conserved process called cap snatching. The recent developments in high-throughput sequencing have made it possible to determine most, if not all, of the capped RNAs snatched by a sNSV. Here, we show that rice stripe tenuivirus (RSV), a plant-infecting sNSV, co-infects Nicotiana benthamiana with two different begomoviruses and snatches capped leader sequences from their mRNAs. By determining the 5′ termini of a single RSV mRNA with high-throughput sequencing, the 5′ ends of almost all the mRNAs of the co-infecting begomoviruses could be identified and mapped on their genomes. The findings in this study provide support for the using of the cap snatching of sNSVs as a tool to map viral TSSs.

Highlights

  • Messenger RNAs in a eukaryotic cell normally contain a co-transcriptionally attached 7-methylguanosine (m7G) cap at their 5′ termini (Shatkin, 1976)

  • Our results provide support for the using of the cap snatching of sense RNA viruses (sNSVs) as a tool for molecular studies

  • To test whether it would be possible to exploit the mechanism of cap snatching for the mapping of begomoviral transcription start sites (TSSs) to their corresponding genome DNA templates, we first verified the occurrence of mixed infections of tenuiviruses and begomoviruses

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Summary

Introduction

Messenger RNAs (mRNAs) in a eukaryotic cell normally contain a co-transcriptionally attached 7-methylguanosine (m7G) cap at their 5′ termini (Shatkin, 1976). Segmented negative-sense RNA viruses (sNSVs) of the order Bunyavirales and the families Orthomyxoviridae and Arenaviridae do not contain MT activity within their polymerases and have evolved a unique mechanism called cap snatching to cap their mRNAs (Decroly et al, 2011; Reich et al, 2014).

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