title: The cellular decapping activators LSm1, Pat1, and Dhh1 control the ratio of subgenomic to genomic Flock House virus RNAs.
abstract: Positive-strand RNA viruses depend on recruited host factors to control critical replication steps. Previously, it was shown that replication of evolutionarily diverse positive-strand RNA viruses, such as hepatitis C virus and brome mosaic virus, depends on host decapping activators LSm1-7, Pat1, and Dhh1 (J. Diez et al., Proc. Natl. Acad. Sci. U. S. A. 97:3913-3918, 2000; A. Mas et al., J. Virol. 80:246 -251, 2006; N. Scheller et al., Proc. Natl. Acad. Sci. U. S. A. 106:13517-13522, 2009). By using a system that allows the replication of the insect Flock House virus (FHV) in yeast, here we show that LSm1-7, Pat1, and Dhh1 control the ratio of subgenomic RNA3 to genomic RNA1 production, a key feature in the FHV life cycle mediated by a long-distance base pairing within RNA1. Depletion of LSM1, PAT1, or DHH1 dramatically increased RNA3 accumulation during replication. This was not caused by differences between RNA1 and RNA3 steady-state levels in the absence of replication. Importantly, coimmunoprecipitation assays indicated that LSm1-7, Pat1, and Dhh1 interact with the FHV RNA genome and the viral polymerase. By using a strategy that allows dissecting different stages of the replication process, we found that LSm1-7, Pat1, and Dhh1 did not affect the early replication steps of RNA1 recruitment to the replication complex or RNA1 synthesis. Furthermore, their function on RNA3/RNA1 ratios was independent of the membrane compartment, where replication occurs and requires ATPase activity of the Dhh1 helicase. Together, these results support that LSm1-7, Pat1, and Dhh1 control RNA3 synthesis. Their described function in mediating cellular mRNP rearrangements suggests a parallel role in mediating key viral RNP transitions, such as the one required to maintain the balance between the alternative FHV RNA1 conformations that control RNA3 synthesis.
authors.affiliation: Department of Experimental and Health Sciences, Universitat Pompeu Fabra, Barcelona, Spain.
authors.forename: Mireia
authors.initials: M
authors.lastname: Giménez-Barcons
authors.forename: Isabel
authors.initials: I
authors.lastname: Alves-Rodrigues
authors.forename: Jennifer
authors.initials: J
authors.lastname: Jungfleisch
authors.forename: Priscilla M
authors.initials: PM
authors.lastname: Van Wynsberghe
authors.forename: Paul
authors.initials: P
authors.lastname: Ahlquist
authors.forename: Juana
authors.initials: J
authors.lastname: Díez
mesh_terms: D053487:DEAD-box RNA Helicases / Q000235:genetics / Q000378:metabolism*; D016679:Genome, Viral; D023281:Genomics; D054884:Host-Pathogen Interactions; D030381:Nodaviridae / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D039381:RNA Cap-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D012367:RNA, Viral / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism* / Q000821:virology*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
chemical_list: C114069:LSM1 protein, S cerevisiae; C103266:PAT1 protein, S cerevisiae; D039381:RNA Cap-Binding Proteins; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins; C445083:DHH1 protein, S cerevisiae; D053487:DEAD-box RNA Helicases
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