title: Visualization of Host-Polerovirus Interaction Topologies Using Protein Interaction Reporter Technology.
abstract: Demonstrating direct interactions between host and virus proteins during infection is a major goal and challenge for the field of virology. Most protein interactions are not binary or easily amenable to structural determination. Using infectious preparations of a polerovirus (Potato leafroll virus [PLRV]) and protein interaction reporter (PIR), a revolutionary technology that couples a mass spectrometric-cleavable chemical cross-linker with high-resolution mass spectrometry, we provide the first report of a host-pathogen protein interaction network that includes data-derived, topological features for every cross-linked site that was identified. We show that PLRV virions have hot spots of protein interaction and multifunctional surface topologies, revealing how these plant viruses maximize their use of binding interfaces. Modeling data, guided by cross-linking constraints, suggest asymmetric packing of the major capsid protein in the virion, which supports previous epitope mapping studies. Protein interaction topologies are conserved with other species in the Luteoviridae and with unrelated viruses in the Herpesviridae and Adenoviridae. Functional analysis of three PLRV-interacting host proteins in planta using a reverse-genetics approach revealed a complex, molecular tug-of-war between host and virus. Structural mimicry and diversifying selection-hallmarks of host-pathogen interactions-were identified within host and viral binding interfaces predicted by our models. These results illuminate the functional diversity of the PLRV-host protein interaction network and demonstrate the usefulness of PIR technology for precision mapping of functional host-pathogen protein interaction topologies.
  
  
  OBJECTIVE
  The exterior shape of a plant virus and its interacting host and insect vector proteins determine whether a virus will be transmitted by an insect or infect a specific host. Gaining this information is difficult and requires years of experimentation. We used protein interaction reporter (PIR) technology to illustrate how viruses exploit host proteins during plant infection. PIR technology enabled our team to precisely describe the sites of functional virus-virus, virus-host, and host-host protein interactions using a mass spectrometry analysis that takes just a few hours. Applications of PIR technology in host-pathogen interactions will enable researchers studying recalcitrant pathogens, such as animal pathogens where host proteins are incorporated directly into the infectious agents, to investigate how proteins interact during infection and transmission as well as develop new tools for interdiction and therapy.
authors.affiliation: Boyce Thompson Institute for Plant Research, Ithaca, New York, USA USDA-Agricultural Research Service, Ithaca, New York, USA.
authors.forename: Stacy L
authors.initials: SL
authors.lastname: DeBlasio
authors.affiliation: Department of Genome Sciences, University of Washington, Seattle, Washington, USA.
authors.forename: Juan D
authors.initials: JD
authors.lastname: Chavez
authors.affiliation: Boyce Thompson Institute for Plant Research, Ithaca, New York, USA Department of Plant Pathology and Plant-Microbe Biology, Cornell University, Ithaca, New York, USA.
authors.forename: Mariko M
authors.initials: MM
authors.lastname: Alexander
authors.affiliation: Boyce Thompson Institute for Plant Research, Ithaca, New York, USA.
authors.forename: John
authors.initials: J
authors.lastname: Ramsey
authors.affiliation: University of Washington Proteomics Resources, Seattle, Washington, USA.
authors.forename: Jimmy K
authors.initials: JK
authors.lastname: Eng
authors.affiliation: Boyce Thompson Institute for Plant Research, Ithaca, New York, USA.
authors.forename: Jaclyn
authors.initials: J
authors.lastname: Mahoney
authors.affiliation: USDA-Agricultural Research Service, Ithaca, New York, USA Department of Plant Pathology and Plant-Microbe Biology, Cornell University, Ithaca, New York, USA.
authors.forename: Stewart M
authors.initials: SM
authors.lastname: Gray
authors.affiliation: Department of Genome Sciences, University of Washington, Seattle, Washington, USA.
authors.forename: James E
authors.initials: JE
authors.lastname: Bruce
authors.affiliation: Boyce Thompson Institute for Plant Research, Ithaca, New York, USA USDA-Agricultural Research Service, Ithaca, New York, USA Department of Plant Pathology and Plant-Microbe Biology, Cornell University, Ithaca, New York, USA mlc68@cornell.edu.
authors.forename: Michelle
authors.initials: M
authors.lastname: Cilia
mesh_terms: D054884:Host-Pathogen Interactions; D052679:Luteoviridae / Q000502:physiology*; D010940:Plant Proteins / Q000378:metabolism; D060066:Protein Interaction Maps; D040901:Proteomics / Q000379:methods*; D014026:Nicotiana; D014764:Viral Proteins / Q000378:metabolism
chemical_list: D010940:Plant Proteins; D014764:Viral Proteins