Interaction of the viral protein genome linked of turnip mosaic potyvirus with the translational eukaryotic initiation factor (iso) 4E of Arabidopsis thaliana using the yeast two-hybrid system.
The yeast LexA interaction trap was used to screen a cDNA library from Arabidopsis thaliana in order to identify proteins that interact with the viral protein genome linked (VPg)-proteinase of turnip mosaic potyvirus. The screen allowed the isolation of four candidate cDNA clones. Clones pHC4, pHC21, and pHC40 were partially sequenced but no homologies to known proteins were found. However, the amino acid sequence deduced from the complete nucleotide sequence of pSW56 revealed that it was the eukaryotic initiation factor (iso) 4E [eIF(iso)4E]. Deletion analysis indicated that the VPg domain was involved in the interaction with the plant protein. Interaction between the viral protein and the cellular protein was confirmed by ELISA-based binding experiments. eIF(iso)4E plays an essential role in the initiation of the translation of capped mRNAs and its association with VPg would point to a role of the viral protein in the translation of the virus.
Wittmann
S
S
Centre de recherche en virologie, Institut Armand-Frappier, Ville de Laval, Québec, Canada.
Chatel
H
H
Fortin
M G
MG
Laliberté
J F
JF
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D002461:Cell Line, Transformed; D004279:DNA, Viral; D039561:Eukaryotic Initiation Factor-4E; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D010448:Peptide Initiation Factors / Q000235:genetics / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D017800:Potyvirus / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae; D017386:Sequence Homology, Amino Acid; D014758:Viral Core Proteins / Q000235:genetics / Q000378:metabolism*
D004279:DNA, Viral; D039561:Eukaryotic Initiation Factor-4E; D010448:Peptide Initiation Factors; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins