title: Eukaryotic initiation factor (eIF) 4F binding to barley yellow dwarf virus (BYDV) 3'-untranslated region correlates with translation efficiency.
abstract: Eukaryotic initiation factor (eIF) 4F binding to mRNA is the first committed step in cap-dependent protein synthesis. Barley yellow dwarf virus (BYDV) employs a cap-independent mechanism of translation initiation that is mediated by a structural BYDV translation element (BTE) located in the 3'-UTR of its mRNA. eIF4F bound the BTE and a translationally inactive mutant with high affinity, thus questioning the role of eIF4F in translation of BYDV. To examine the effects of eIF4F in BYDV translation initiation, BTE mutants with widely different in vitro translation efficiencies ranging from 5 to 164% compared with WT were studied. Using fluorescence anisotropy to obtain quantitative data, we show 1) the equilibrium binding affinity (complex stability) correlated well with translation efficiency, whereas the "on" rate of binding did not; 2) other unidentified proteins or small molecules in wheat germ extract prevented eIF4F binding to mutant BTE but not WT BTE; 3) BTE mutant-eIF4F interactions were found to be both enthalpically and entropically favorable with an enthalpic contribution of 52-90% to ΔG° at 25 °C, suggesting that hydrogen bonding contributes to stability; and 4) in contrast to cap-dependent and tobacco etch virus internal ribosome entry site interaction with eIF4F, poly(A)-binding protein did not increase eIF4F binding. Further, the eIF4F bound to the 3' BTE with higher affinity than for either m(7)G cap or tobacco etch virus internal ribosome entry site, suggesting that the 3' BTE may play a role in sequestering host cell initiation factors and possibly regulating the switch from replication to translation.
authors.affiliation: From the Department of Chemistry, Hunter College and the Graduate Center of the City University of New York, New York, New York 10065.
authors.forename: Bidisha
authors.initials: B
authors.lastname: Banerjee
authors.forename: Dixie J
authors.initials: DJ
authors.lastname: Goss
mesh_terms: D020413:3' Untranslated Regions; D039562:Eukaryotic Initiation Factor-4F / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017793:Luteovirus / Q000737:chemistry* / Q000502:physiology; D009154:Mutation; D010442:Peptide Chain Initiation, Translational / Q000502:physiology*; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication / Q000502:physiology
chemical_list: D020413:3' Untranslated Regions; D039562:Eukaryotic Initiation Factor-4F; D010940:Plant Proteins; D012367:RNA, Viral; D011994:Recombinant Proteins
keywords: Fluorescence; Plant Viruses; Protein Synthesis; Protein-Nucleic Acid Interaction; RNA Viruses; eIF4F