Efficient translation of rotavirus mRNA requires simultaneous interaction of NSP3 with the eukaryotic translation initiation factor eIF4G and the mRNA 3' end.
In contrast to the vast majority of cellular proteins, rotavirus proteins are translated from capped but nonpolyadenylated mRNAs. The viral nonstructural protein NSP3 specifically binds the 3'-end consensus sequence of viral mRNAs and interacts with the eukaryotic translation initiation factor eIF4G. Here we show that expression of NSP3 in mammalian cells allows the efficient translation of virus-like mRNA. A synergistic effect between the cap structure and the 3' end of rotavirus mRNA was observed in NSP3-expressing cells. The enhancement of viral mRNA translation by NSP3 was also observed in a rabbit reticulocyte lysate translation system supplemented with recombinant NSP3. The use of NSP3 mutants indicates that its RNA- and eIF4G-binding domains are both required to enhance the translation of viral mRNA. The results reported here show that NSP3 forms a link between viral mRNA and the cellular translation machinery and hence is a functional analogue of cellular poly(A)-binding protein.
Vende
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P
Laboratoire de Virologie et Immunologie Moléculaires INRA, C.R.J.J., 78352 Jouy-en-Josas Cedex, France.
Piron
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M
Castagné
N
N
Poncet
D
D
D000818:Animals; D002460:Cell Line; D039603:Eukaryotic Initiation Factor-4G; D010448:Peptide Initiation Factors / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D011061:Poly A / Q000378:metabolism; D014176:Protein Biosynthesis; D012315:RNA Caps / Q000378:metabolism; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011817:Rabbits; D011994:Recombinant Proteins; D012156:Reticulocytes / Q000378:metabolism; D012401:Rotavirus / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism*
D039603:Eukaryotic Initiation Factor-4G; C082740:NSP3 protein, Rotavirus; D010448:Peptide Initiation Factors; D012315:RNA Caps; D012333:RNA, Messenger; D012367:RNA, Viral; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins; D011061:Poly A