Reovirus low-density particles package viral and cellular RNA
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA689770 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR13360400_1.fastq.gz | Fastqsanger.gz | |||
| SRR13360400_2.fastq.gz | Fastqsanger.gz | |||
| SRR13360401_1.fastq.gz | Fastqsanger.gz | |||
| SRR13360401_2.fastq.gz | Fastqsanger.gz | |||
| SRR13360402_1.fastq.gz | Fastqsanger.gz |
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Project description:Packaging of segmented, double-stranded RNA viral genomes requires coordination of multiple viral proteins and RNA segments. For mammalian orthoreovirus (reovirus), evidence suggests either all ten or zero viral RNA segments are simultaneously packaged in a highly coordinated process hypothesized to exclude host RNA. Accordingly, reovirus generates genome-containing virions and “genomeless” top component particles. However, despite ostensibly lacking the genome, top component particles maintain a low level of infectivity. Whether reovirus particles can package host RNA is unknown. To gain insight into reovirus packaging potential and mechanisms, we employed next-generation RNA-sequencing to define the viral and host RNA content of purified reovirus virions and top component particles. Reovirus top component particles contained double-stranded viral RNA segments in similar proportions but at reduced levels compared to virions. Top component particles also were enriched for numerous host RNAs, especially short, non-polyadenylated transcripts, that differed by reovirus strain, independent of the viral polymerase. In contrast, virions were enriched for very few host RNAs. Collectively, these findings indicate that genome packaging into reovirus virions is exquisitely selective, while incorporation of host RNAs into top component particles is more promiscuous or differentially selective and may contribute to or result from inefficient viral RNA packaging.
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Project description:Packaging of segmented, double-stranded RNA viral genomes requires coordination of viral proteins and RNA segments. For mammalian orthoreovirus (reovirus), evidence suggests either all ten or zero viral RNA segments are simultaneously packaged in a highly coordinated process hypothesized to exclude host RNA. Accordingly, reovirus generates genome-containing virions and "genomeless" top component particles. Whether reovirus virions or top component particles package host RNA is unknown. To gain insight into reovirus packaging potential and mechanisms, we employed next-generation RNA-sequencing to define the RNA content of enriched reovirus particles. Reovirus virions exclusively packaged viral double-stranded RNA. In contrast, reovirus top component particles contained similar proportions but reduced amounts of viral double-stranded RNA and were selectively enriched for numerous host RNA species, especially short, non-polyadenylated transcripts. Host RNA selection was not dependent on RNA abundance in the cell, and specifically enriched host RNAs varied for two reovirus strains and were not selected solely by the viral RNA polymerase. Collectively, these findings indicate that genome packaging into reovirus virions is exquisitely selective, while incorporation of host RNAs into top component particles is differentially selective and may contribute to or result from inefficient viral RNA packaging.
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Project description:Infection of the chestnut blight fungus, Cryphonectria parasitica, by hypovirus CHV1-EP713 or by reoviruses MYRV1-Cp9B21 or MYRV2-CpC18 results in reduced fungal virulence (hypovirulence). However, additional phenotypic changes caused by the two groups of mycoviruses are quite different. CHV1-EP713 infection results in depressed pigmentation and conidiation while reovirus infection has little effect on these processes. We now report that loss of female fertility and resulting absence of virus transmission through sexual spores observed after hypovirus infection was not observed for reovirus infected C. parasitica. Reovirus-infected strains were male and female fertile and able to transmit virus to ascospore progeny at a high rate when serving as a female parent. Consistent with this result, real-time RT-PCR revealed that expression of two genes involved in sexual reproduction, the pheromone precursor gene, mf2-1 and yeast Ste12-like transcriptional factor gene, Cpst12, were less reduced in reovirus-infected strains than in hypovirus-infected strains. Analysis with a custom microarray cDNA chip containing EST clones representing ca 2,200 unique C. parasitica genes identified 140 and 128 host genes that were responsive to MYRV1-Cp9B21 and MYRV2-CpC18 infection, respectively. Comparison of these virus-responsive genes revealed an overlap of 85 genes even though the overall degree of nucleotide sequence identity of the two reoviruses is less than 50%. Significantly, 84 of the 85 genes were altered in the same direction. Further comparison revealed that 51 % and 48% of the genes that were responsive to reoviruses MYRV1-Cp9B21 and MYRV2-CpC18 infection were also responsive to CHV1-EP713 infection. Finally, similar to results reported for hypovirus infection, a high percentage (59% and 66%) of the mycoreovirus responsive genes were also differentially expressed following disruption of the cellular G-protein signal transduction pathway. These data support the hypothesis that hypovirus and reovirus infections perturb common and specific C. parasitica regulatory pathways to cause hypovirulence and distinct sets of phenotypic changes. Keywords: Mycoreovirus, hypovirus, gene expression, microarray.
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