Journal
VIRUSES-BASEL
Volume 13, Issue 11, Pages -Publisher
MDPI
DOI: 10.3390/v13112190
Keywords
eukaryotic translation elongation factor 1A (eEF1A); host factor; nanoLC-MS/MS; SDS-PAGE-based proteomics; pro-viral factor; ribonucleoproteins (RNP); susceptibility factor; Tomato spotted wilt orthotospovirus (TSWV)
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Funding
- Dutch science foundation (NWO-STW) [14948]
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The study analyzed the protein composition of ribonucleoprotein complexes (RNPs) of Tomato spotted wilt orthotospovirus (TSWV) and found that plant RNPs were enriched in proteins related to sugar and phosphate transport, cellular stress responses, while yeast-derived viral RNPs predominantly contained proteins implicated in RNA processing and ribosome biogenesis. This suggests a recruitment of translational machinery to viral RNPs in yeast. Furthermore, gene silencing experiments identified four host factors important for systemic spread of TSWV and disease symptom development.
The tripartite genome of the negative-stranded RNA virus Tomato spotted wilt orthotospovirus (TSWV) is assembled, together with two viral proteins, the nucleocapsid protein and the RNA-dependent RNA polymerase, into infectious ribonucleoprotein complexes (RNPs). These two viral proteins are, together, essential for viral replication and transcription, yet our knowledge on the host factors supporting these two processes remains limited. To fill this knowledge gap, the protein composition of viral RNPs collected from TSWV-infected Nicotiana benthamiana plants, and of those collected from a reconstituted TSWV replicon system in the yeast Saccharomyces cerevisiae, was analysed. RNPs obtained from infected plant material were enriched for plant proteins implicated in (i) sugar and phosphate transport and (ii) responses to cellular stress. In contrast, the yeast-derived viral RNPs primarily contained proteins implicated in RNA processing and ribosome biogenesis. The latter suggests that, in yeast, the translational machinery is recruited to these viral RNPs. To examine whether one of these cellular proteins is important for a TSWV infection, the corresponding N. benthamiana genes were targeted for virus-induced gene silencing, and these plants were subsequently challenged with TSWV. This approach revealed four host factors that are important for systemic spread of TSWV and disease symptom development.
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