title: Model System-Guided Protein Interaction Mapping for Virus Isolated from Phloem Tissue.
abstract: Phloem localization of plant viruses is advantageous for acquisition by sap-sucking vectors but hampers host-virus protein interaction studies. In this study, Potato leafroll virus (PLRV)-host protein complexes were isolated from systemically infected potato, a natural host of the virus. Comparing two different co-immunoprecipitation (co-IP) support matrices coupled to mass spectrometry (MS), we identified 44 potato proteins and one viral protein (P1) specifically associated with virus isolated from infected phloem. An additional 142 proteins interact in complex with virus at varying degrees of confidence. Greater than 80% of these proteins were previously found to form high confidence interactions with PLRV isolated from the model host Nicotiana benthamiana. Bioinformatics revealed that these proteins are enriched for functions related to plasmodesmata, organelle membrane transport, translation, and mRNA processing. Our results show that model system proteomics experiments are extremely valuable for understanding protein interactions regulating infection in recalcitrant pathogens such as phloem-limited viruses.
authors.affiliation: Agricultural Research Service, USDA , Ithaca, New York 14853, United States.|Boyce Thompson Institute for Plant Research , Ithaca, New York 14853, United States.
authors.forename: Stacy L
authors.initials: SL
authors.lastname: DeBlasio
authors.affiliation: Department of Genome Sciences, University of Washington , Seattle Washington 98109, United States.
authors.forename: Richard S
authors.initials: RS
authors.lastname: Johnson
authors.affiliation: Department of Genome Sciences, University of Washington , Seattle Washington 98109, United States.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: MacCoss
authors.affiliation: Agricultural Research Service, USDA , Ithaca, New York 14853, United States.|Department of Plant Pathology and Plant-Microbe Biology, Cornell University , Ithaca, New York 14853, United States.
authors.forename: Stewart M
authors.initials: SM
authors.lastname: Gray
authors.affiliation: Agricultural Research Service, USDA , Ithaca, New York 14853, United States.|Boyce Thompson Institute for Plant Research , Ithaca, New York 14853, United States.|Department of Plant Pathology and Plant-Microbe Biology, Cornell University , Ithaca, New York 14853, United States.
authors.forename: Michelle
authors.initials: M
authors.lastname: Cilia
mesh_terms: D019295:Computational Biology; D054884:Host-Pathogen Interactions; D052585:Phloem / Q000821:virology*; D010940:Plant Proteins / Q000378:metabolism; D010942:Plant Viruses / Q000737:chemistry; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D011198:Solanum tuberosum / Q000737:chemistry / Q000821:virology; D014764:Viral Proteins / Q000378:metabolism
chemical_list: D010940:Plant Proteins; D014764:Viral Proteins
keywords: Luteoviridae; Potato leafroll virus; SAINT; Solanum tuberosum; co-immunoprecipitation; high-resolution mass spectrometry; host−pathogen interactions; model system; phloem-limited; polerovirus