In vitro and in vivo mapping of the Prunus necrotic ringspot virus coat protein C-terminal dimerization domain by bimolecular fluorescence complementation.
Interactions between viral proteins are critical for virus viability. Bimolecular fluorescent complementation (BiFC) technique determines protein interactions in real-time under almost normal physiological conditions. The coat protein (CP) of Prunus necrotic ringspot virus is required for multiple functions in its replication cycle. In this study, the region involved in CP dimerization has been mapped by BiFC in both bacteria and plant tissue. Full-length and C-terminal deleted forms of the CP gene were fused in-frame to the N- and C-terminal fragments of the yellow fluorescent protein. The BiFC analysis showed that a domain located between residues 9 and 27 from the C-end plays a critical role in dimerization. The importance of this C-terminal region in dimer formation and the applicability of the BiFC technique to analyse viral protein interactions are discussed.
Aparicio
Frederic
F
Instituto de Biología Molecular y Celular de Plantas, Universidad Politécnica de Valencia (CSIC), Av. de los Naranjos s/n, 46022 Valencia, Spain.
Sánchez-Navarro
Jesús A
JA
Instituto de Biología Molecular y Celular de Plantas, Universidad Politécnica de Valencia (CSIC), Av. de los Naranjos s/n, 46022 Valencia, Spain.
Pallás
Vicente
V
Instituto de Biología Molecular y Celular de Plantas, Universidad Politécnica de Valencia (CSIC), Av. de los Naranjos s/n, 46022 Valencia, Spain.
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D036022:Capsid Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019281:Dimerization; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D019179:Ilarvirus / Q000737:chemistry / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D010935:Plant Diseases / Q000821:virology*; D027861:Prunus / Q000821:virology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000821:virology
D001426:Bacterial Proteins; D036022:Capsid Proteins; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; C054562:yellow fluorescent protein, Bacteria