Complex formation between potyvirus VPg and translation eukaryotic initiation factor 4E correlates with virus infectivity.
The interaction between the viral protein linked to the genome (VPg) of turnip mosaic potyvirus (TuMV) and the translation eukaryotic initiation factor eIF(iso)4E of Arabidopsis thaliana has previously been reported. eIF(iso)4E binds the cap structure (m(7)GpppN, where N is any nucleotide) of mRNAs and has an important role in the regulation in the initiation of translation. In the present study, it was shown that not only did VPg bind eIF(iso)4E but it also interacted with the eIF4E isomer of A. thaliana as well as with eIF(iso)4E of Triticum aestivum (wheat). The interaction domain on VPg was mapped to a stretch of 35 amino acids, and substitution of an aspartic acid residue found within this region completely abolished the interaction. The cap analogue m(7)GTP, but not GTP, inhibited VPg-eIF(iso)4E complex formation, suggesting that VPg and cellular mRNAs compete for eIF(iso)4E binding. The biological significance of this interaction was investigated. Brassica perviridis plants were infected with a TuMV infectious cDNA (p35Tunos) and p35TuD77N, a mutant which contained the aspartic acid substitution in the VPg domain that abolished the interaction with eIF(iso)4E. After 20 days, plants bombarded with p35Tunos showed viral symptoms, while plants bombarded with p35TuD77N remained symptomless. These results suggest that VPg-eIF(iso)4E interaction is a critical element for virus production.
Léonard
S
S
Centre de Microbiologie et Biotechnologie, INRS-Institut Armand-Frappier, Ville de Laval, Québec, Canada H7V 1B7.
Plante
D
D
Wittmann
S
S
Daigneault
N
N
Fortin
M G
MG
Laliberté
J F
JF
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D017360:Arabidopsis / Q000378:metabolism / Q000821:virology*; D001224:Aspartic Acid / Q000235:genetics; D001667:Binding, Competitive; D004797:Enzyme-Linked Immunosorbent Assay; D039561:Eukaryotic Initiation Factor-4E; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010448:Peptide Initiation Factors / Q000378:metabolism*; D017800:Potyvirus / Q000378:metabolism / Q000472:pathogenicity*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012315:RNA Caps / Q000378:metabolism; D020539:Sequence Analysis, Protein; D020798:Two-Hybrid System Techniques; D014758:Viral Core Proteins / Q000378:metabolism*; D014774:Virulence
D039561:Eukaryotic Initiation Factor-4E; D010448:Peptide Initiation Factors; D012315:RNA Caps; D014758:Viral Core Proteins; D001224:Aspartic Acid