title: The helper-component protease transmission factor of tobacco etch potyvirus binds specifically to an aphid ribosomal protein homologous to the laminin receptor precursor.
abstract: Potyviruses are plant pathogens transmitted by aphids in a non-persistent manner. During transmission, the virus-encoded factor helper-component protease (HCPro) is presumed to act as a molecular bridge, mediating the reversible retention of virions to uncharacterized binding sites in the vector mouthparts. Whilst the predicted interaction between HCPro and the coat protein (CP) of virions has been confirmed experimentally, the characterization of putative HCPro-specific receptors in aphids has remained elusive, with the exception of a report that described binding of HCPro of zucchini yellow mosaic virus to several cuticle proteins. To identify other aphid components that could play a role during transmission, this study used purified HCPro of tobacco etch virus (TEV) in far-Western blotting assays as bait to select interactors among proteins extracted from aphid heads. With this approach, new HCPro-interacting proteins were found, and several were identified after mass spectrometry analysis and searches in databases dedicated to aphid sequences. Among these interactors, a ribosomal protein S2 (RPS2) was chosen for further investigation due to its homology with the laminin receptor precursor, known to act as the receptor of several viruses. The specific interaction between RPS2 and TEV HCPro was confirmed after cloning and heterologous expression of the corresponding Myzus persicae gene. The possible involvement of RPS2 in the transmission process was further suggested by testing a variant of HCPro that was non-functional for transmission due to a mutation in the conserved KITC motif (EITC variant). This variant retained its ability to bind CP but failed to interact with RPS2.
authors.affiliation: Departamento de Biología de Plantas, Centro de Investigaciones Biológicas (CIB, CSIC), Ramiro de Maeztu 9, 28040 Madrid, Spain.
authors.forename: Lourdes
authors.initials: L
authors.lastname: Fernández-Calvino
authors.forename: Elisa
authors.initials: E
authors.lastname: Goytia
authors.forename: Dionisio
authors.initials: D
authors.lastname: López-Abella
authors.forename: Ana
authors.initials: A
authors.lastname: Giner
authors.forename: María
authors.initials: M
authors.lastname: Urizarna
authors.forename: Lluisa
authors.initials: L
authors.lastname: Vilaplana
authors.forename: Juan José
authors.initials: JJ
authors.lastname: López-Moya
mesh_terms: D000818:Animals; D001042:Aphids / Q000821:virology; D026481:Blotting, Far-Western; D019476:Insect Proteins / Q000378:metabolism*; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism*; D010935:Plant Diseases / Q000821:virology; D017800:Potyvirus / Q000502:physiology*; D011485:Protein Binding; D025941:Protein Interaction Mapping; D017474:Receptors, Laminin / Q000235:genetics; D012269:Ribosomal Proteins / Q000235:genetics / Q000378:metabolism*; D014026:Nicotiana / Q000821:virology; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D019476:Insect Proteins; D050505:Mutant Proteins; D017474:Receptors, Laminin; D012269:Ribosomal Proteins; D014764:Viral Proteins; C030004:ribosomal protein S2; D010447:Peptide Hydrolases