Title: | Spectroscopic Investigation of the Kinetic Mechanism Involved in the Association of Potyviral VPg with the Host Plant Translation Initiation Factor eIF4E |
Author(s): | Walter J; Barra A; Charon J; Tavert-Roudet G; Michon T; |
Address: | "INRAE, Biologie du Fruit et Pathologie, University of Bordeaux, UMR 1332, F-33140 Villenave d'Ornon, France. Faculty of Sciences, University of Sydney, Charles Perkins Center D17, Camperdown Campus, Sydney, NSW 2006, Australia" |
ISSN/ISBN: | 1422-0067 (Electronic) 1422-0067 (Linking) |
Abstract: | "The infectious cycle of potyviruses requires the formation of a complex between the viral genome-linked protein VPg and the host eukaryotic translation initiation factor 4E, eIF4E. Mutations associated with plant resistance to potyviruses were previously mapped at the eIF4E surface, while on the virus side, mutations leading to plant resistance breaking were identified within the VPg. In the present study, fluorescence spectroscopy was used to probe the contribution of the VPg intrinsically disordered region bearing amino acids determinant of the resistance breaking, to the VPg-eIF4E binding mechanism. Synthetic peptides encompassing the VPg(88-120) central region were found to tightly bind to eIF4E. Fluorescence energy transfer experiments show that, upon binding to eIF4E, the N and C termini of the VPg(88-111) fragment move closer to one another, at a distance compatible with a alpha-helix folding. When the VPg(112-120) region, which contains amino acids associated with resistance breakdown, is appended to VPg(88-111), the complex formation with eIF4E switches from a single-step to a two-step kinetic model. This study revisits a recent investigation of the VPg-eIF4E complex by specifying the contribution of the VPg central helix and its appended disordered region to VPg association with eIF4E" |
Keywords: | "Amino Acid Sequence/genetics Binding Sites/genetics Eukaryotic Initiation Factor-4E/*chemistry/genetics Genome, Viral/genetics Host-Pathogen Interactions/genetics Kinetics Plant Defense Against Herbivory/genetics Plant Diseases/*genetics/virology Plant Pr;" |
Notes: | "MedlineWalter, Jocelyne Barra, Amandine Charon, Justine Tavert-Roudet, Genevieve Michon, Thierry eng Switzerland 2020/08/09 Int J Mol Sci. 2020 Aug 5; 21(16):5618. doi: 10.3390/ijms21165618" |