[Complex formation between recombinant protein VPg of potato virus Y and heterologous eukaryotic translation initiation factors eIF4E].
Genomes of some positive-strand RNA viruses do not contain cap-structure, but instead their 5'-end is covalently linked to a viral protein called VPg. Complex formation between VPg and cellular translation initiation factors (eIFs) has been extensively studied in the context of the model of this complex involvement in virus mRNA translation initiation and cellular protein translation shut down in infected cells. The potato virus (PVY) VPg was expressed in bacterial and baculovirus systems in order to investigate its binding capacity to wheat eIF4E and its isoform. Both purified recombinant eIF4E and eIF(iso)4E were identified in vitro as binding partners of the purified recombinant VPg by using affinity chromatography, as well in vivo by coexpressing of recombinant VPg and eIFs in insect cells with following complex purification using affinity chromatography. Besides it was shown that PVY VPg also formed a complex with endogenous insect eIF4E in vivo. PVY VPg interaction with eIF4E of wheat (non permissive plant for PVY), and also with so evolutionary distant partner as insect eIF4E suggests the conservation of general structural features of eIF4E implicated in the formation of the complex with VPg.
Strokovskaia
L I
LI
Grzela
R
R
Chroboczek
J
J
Kikhno
I M
IM
Chashchina
L I
LI
Solomko
A P
AP
D000818:Animals; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D039561:Eukaryotic Initiation Factor-4E / Q000235:genetics*; D016679:Genome, Viral; D010957:Plasmids; D017800:Potyvirus / Q000235:genetics* / Q000378:metabolism; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics*; D012261:Ribonucleoproteins / Q000235:genetics*; D011198:Solanum tuberosum / Q000378:metabolism / Q000821:virology; D018411:Spodoptera / Q000378:metabolism / Q000821:virology; D014908:Triticum / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000235:genetics*
D039561:Eukaryotic Initiation Factor-4E; D011994:Recombinant Proteins; D012261:Ribonucleoproteins; D017361:Viral Nonstructural Proteins; C102206:genome-linked viral protein, potato virus Y