title: Phosphatidic acid produced by phospholipase D promotes RNA replication of a plant RNA virus.
abstract: Eukaryotic positive-strand RNA [(+)RNA] viruses are intracellular obligate parasites replicate using the membrane-bound replicase complexes that contain multiple viral and host components. To replicate, (+)RNA viruses exploit host resources and modify host metabolism and membrane organization. Phospholipase D (PLD) is a phosphatidylcholine- and phosphatidylethanolamine-hydrolyzing enzyme that catalyzes the production of phosphatidic acid (PA), a lipid second messenger that modulates diverse intracellular signaling in various organisms. PA is normally present in small amounts (less than 1% of total phospholipids), but rapidly and transiently accumulates in lipid bilayers in response to different environmental cues such as biotic and abiotic stresses in plants. However, the precise functions of PLD and PA remain unknown. Here, we report the roles of PLD and PA in genomic RNA replication of a plant (+)RNA virus, Red clover necrotic mosaic virus (RCNMV). We found that RCNMV RNA replication complexes formed in Nicotiana benthamiana contained PLDα and PLDβ. Gene-silencing and pharmacological inhibition approaches showed that PLDs and PLDs-derived PA are required for viral RNA replication. Consistent with this, exogenous application of PA enhanced viral RNA replication in plant cells and plant-derived cell-free extracts. We also found that a viral auxiliary replication protein bound to PA in vitro, and that the amount of PA increased in RCNMV-infected plant leaves. Together, our findings suggest that RCNMV hijacks host PA-producing enzymes to replicate.
authors.affiliation: Laboratory of Plant Pathology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
authors.forename: Kiwamu
authors.initials: K
authors.lastname: Hyodo
authors.affiliation: Institute for Enzyme Research, University of Tokushima, Tokushima, Japan.
authors.forename: Takako
authors.initials: T
authors.lastname: Taniguchi
authors.affiliation: Laboratory of Marine Bioproducts Technology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Manabe
authors.affiliation: Laboratory of Plant Pathology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
authors.forename: Masanori
authors.initials: M
authors.lastname: Kaido
authors.affiliation: Laboratory of Plant Pathology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Mise
authors.affiliation: Laboratory of Marine Bioproducts Technology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Sugawara
authors.affiliation: Institute for Enzyme Research, University of Tokushima, Tokushima, Japan.
authors.forename: Hisaaki
authors.initials: H
authors.lastname: Taniguchi
authors.affiliation: Laboratory of Plant Pathology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Okuno
mesh_terms: D015153:Blotting, Western; D020868:Gene Silencing; D047468:Immunoprecipitation; D010712:Phosphatidic Acids / Q000378:metabolism*; D010739:Phospholipase D / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D018515:Plant Leaves / Q000821:virology*; D012333:RNA, Messenger / Q000235:genetics; D018749:RNA, Plant / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014026:Nicotiana / Q000821:virology*; D019183:Tombusviridae / Q000502:physiology*; D014779:Virus Replication
chemical_list: D010712:Phosphatidic Acids; D012333:RNA, Messenger; D018749:RNA, Plant; D034741:RNA, Small Interfering; D012367:RNA, Viral; D010739:Phospholipase D