title: Production of plum pox virus HC-Pro functionally active for aphid transmission in a transient-expression system.
abstract: Potyviruses are non-persistently transmitted by aphid vectors with the assistance of a viral accessory factor known as helper component (HC-Pro), a multifunctional protein that is also involved in many other essential processes during the virus infection cycle. A transient Agrobacterium-mediated expression system was used to produce Plum pox virus (PPV) HC-Pro in Nicotiana benthamiana leaves from constructs that incorporated the 5' region of the genome, yielding high levels of HC-Pro in agroinfiltrated leaves. The expressed PPV HC-Pro was able to assist aphid transmission of purified virus particles in a sequential feeding assay, and to complement transmission-defective variants of the virus. Also, HC-Pro of a second potyvirus, Tobacco etch virus (TEV), was expressed and found to be functional for aphid transmission. These results show that this transient system can be useful for production of functionally active HC-Pro in potyviruses, and the possible uses of this approach to study the mechanism of transmission are discussed.
authors.affiliation: Departamento de Biología de Plantas, Centro de Investigaciones Biológicas (CIB, CSIC), Ramiro de Maeztu 9, 28040 Madrid, Spain.
authors.forename: Elisa
authors.initials: E
authors.lastname: Goytia
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.affiliation: Departamento de Biología de Plantas, Centro de Investigaciones Biológicas (CIB, CSIC), Ramiro de Maeztu 9, 28040 Madrid, Spain.
authors.forename: Belén
authors.initials: B
authors.lastname: Martínez-García
authors.affiliation: Departamento de Biología de Plantas, Centro de Investigaciones Biológicas (CIB, CSIC), Ramiro de Maeztu 9, 28040 Madrid, Spain.
authors.forename: Dionisio
authors.initials: D
authors.lastname: López-Abella
authors.affiliation: Consorcio CSIC-IRTA, Instituto de Biología Molecular de Barcelona (IBMB, CSIC), Jordi Girona 18-26, 08034 Barcelona, Spain.|Departamento de Biología de Plantas, Centro de Investigaciones Biológicas (CIB, CSIC), Ramiro de Maeztu 9, 28040 Madrid, Spain.
authors.forename: Juan José
authors.initials: JJ
authors.lastname: López-Moya
mesh_terms: D000818:Animals; D001042:Aphids / Q000821:virology*; D003546:Cysteine Endopeptidases / Q000096:biosynthesis* / Q000502:physiology; D007303:Insect Vectors / Q000821:virology*; D010935:Plant Diseases / Q000821:virology*; D018515:Plant Leaves / Q000378:metabolism; D017801:Plum Pox Virus / Q000737:chemistry / Q000472:pathogenicity*; D017800:Potyvirus / Q000737:chemistry / Q000472:pathogenicity; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis; D012231:Rhizobium / Q000378:metabolism; D014026:Nicotiana / Q000378:metabolism; D014764:Viral Proteins / Q000096:biosynthesis* / Q000502:physiology; D014774:Virulence
chemical_list: D011994:Recombinant Proteins; D014764:Viral Proteins; D003546:Cysteine Endopeptidases; C072775:HC-Pro protein, potyvirus