A new plant protein interacts with eIF3 and 60S to enhance virus-activated translation re-initiation

• Published in: The EMBO journal Vol. 28; no. 20; pp. 3171 - 3184
• Main Authors: Thiébeauld, Odon, Schepetilnikov, Mikhail, Park, Hyun-Sook, Geldreich, Angèle, Kobayashi, Kappei, Keller, Mario, Hohn, Thomas, Ryabova, Lyubov A
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 21.10.2009; Blackwell Publishing Ltd; EMBO Press; Nature Publishing Group
• Subjects: Arabidopsis Proteins - genetics; Arabidopsis Proteins - metabolism; Botany; Cauliflower mosaic virus (CaMV); Caulimovirus - genetics; Caulimovirus - metabolism; Caulimovirus - physiology; Cellular Biology; eIF3; Eukaryotic Initiation Factor-3 - metabolism; Gene Expression Regulation, Plant; large ribosomal subunit (60S); Life Sciences; Models, Biological; Molecular biology; Mutation; Polymerase Chain Reaction; Polyribosomes - metabolism; Protein Binding - genetics; Protein Binding - physiology; Protein Biosynthesis - genetics; Protein Biosynthesis - physiology; Proteins; re-initiation supporting protein (RISP); Ribosome Subunits, Large, Eukaryotic - metabolism; Ribosome Subunits, Small, Eukaryotic - metabolism; transactivator viroplasmin (TAV); Two-Hybrid System Techniques; Viral Proteins - genetics; Viral Proteins - metabolism; Viruses
• ISSN: 0261-4189; 1460-2075
• DOI: 10.1038/emboj.2009.256

The plant viral re‐initiation factor transactivator viroplasmin (TAV) activates translation of polycistronic mRNA by a re‐initiation mechanism involving translation initiation factor 3 (eIF3) and the 60S ribosomal subunit (60S). QJ;Here, we report a new plant factor—re‐initiation supporting protein (RISP)—that enhances TAV function in re‐initiation. RISP interacts physically with TAV in vitro and in vivo. Mutants defective in interaction are less active, or inactive, in transactivation and viral amplification. RISP alone can serve as a scaffold protein, which is able to interact with eIF3 subunits a/c and 60S, apparently through the C‐terminus of ribosomal protein L24. RISP pre‐bound to eIF3 binds 40S, suggesting that RISP enters the translational machinery at the 43S formation step. RISP, TAV and 60S co‐localize in epidermal cells of infected plants, and eIF3–TAV–RISP–L24 complex formation can be shown in vitro. These results suggest that RISP and TAV bridge interactions between eIF3‐bound 40S and L24 of 60S after translation termination to ensure 60S recruitment during repetitive initiation events on polycistronic mRNA; RISP can thus be considered as a new component of the cell translation machinery.