PT  - JOURNAL ARTICLE
AU  - Sebastian Strauss
AU  - Alexander Borodavka
AU  - Guido Papa
AU  - Daniel Desiró
AU  - Florian Schueder
AU  - Ralf Jungmann
TI  - Principles of RNA recruitment to viral ribonucleoprotein condensates in a segmented dsRNA virus
AID  - 10.1101/2021.03.22.435476
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.03.22.435476
4099  - http://biorxiv.org/content/early/2021/03/22/2021.03.22.435476.short
4100  - http://biorxiv.org/content/early/2021/03/22/2021.03.22.435476.full
AB  - Rotaviruses transcribe eleven distinct protein-coding RNAs that must be stoichiometrically co-packaged prior to their replication to make an infectious virion. During infection, rotavirus transcripts accumulate in cytoplasmic ribonucleoprotein (RNP) condensates, termed viroplasms. Understanding the mechanisms of viroplasm assembly and RNA enrichment within is crucial to gaining greater insight into their function and stoichiometric assortment of individual transcripts. We analysed the subcellular distribution of individual RV transcripts and viroplasm transcriptome by combining multiplexed DNA-barcoded single-molecule RNA FISH of infected cells. Using DNA-PAINT microscopy, we provide evidence of the early onset of viral transcript oligomerisation that occurs prior to the formation of viroplasms. We demonstrate that viral sequences lacking the conserved terminal regions fail to undergo enrichment in rotavirus RNP condensates. We show that individual viral transcripts exhibit variable propensities to partition into viroplasms, irrespective of their absolute numbers in cells, suggesting a selective RNA enrichment mechanism distinct from other known cellular RNP granules. We suggest that rotavirus replication factories represent unique RNP condensates enriched in eleven types of cognate transcripts that may facilitate the assembly of a multi-segmented RNA genome.Competing Interest StatementThe authors have declared no competing interest.