title: A cell cycle-dependent protein serves as a template-specific translation initiation factor.
abstract: Cap-independent translation initiation on picornavirus mRNAs is mediated by an internal ribosomal entry site (IRES) in the 5' untranslated region (5' UTR) and requires both eukaryotic initiation factors (eIFs) and IRES-specific cellular trans-acting factors (ITAFs). We show here that the requirements for trans-acting factors differ between related picornavirus IRESs and can account for cell type-specific differences in IRES function. The neurovirulence of Theiler's murine encephalomyelitis virus (TMEV; GDVII strain) was completely attenuated by substituting its IRES by that of foot-and-mouth disease virus (FMDV). Reconstitution of initiation using fully fractionated translation components indicated that 48S complex formation on both IRESs requires eIF2, eIF3, eIF4A, eIF4B, eIF4F, and the pyrimidine tract-binding protein (PTB) but that the FMDV IRES additionally requires ITAF(45), also known as murine proliferation-associated protein (Mpp1), a proliferation-dependent protein that is not expressed in murine brain cells. ITAF(45) did not influence assembly of 48S complexes on the TMEV IRES. Specific binding sites for ITAF(45), PTB, and a complex of the eIF4G and eIF4A subunits of eIF4F were mapped onto the FMDV IRES, and the cooperative function of PTB and ITAF(45) in promoting stable binding of eIF4G/4A to the IRES was characterized by chemical and enzymatic footprinting. Our data indicate that PTB and ITAF(45) act as RNA chaperones that control the functional state of a particular IRES and that their cell-specific distribution may constitute a basis for cell-specific translational control of certain mRNAs.
authors.affiliation: Institute of Poliomyelitis and Viral Encephalitides, Russian Academy of Medical Sciences, Moscow 142782, Russia.
authors.forename: E V
authors.initials: EV
authors.lastname: Pilipenko
authors.forename: T V
authors.initials: TV
authors.lastname: Pestova
authors.forename: V G
authors.initials: VG
authors.lastname: Kolupaeva
authors.forename: E V
authors.initials: EV
authors.lastname: Khitrina
authors.forename: A N
authors.initials: AN
authors.lastname: Poperechnaya
authors.forename: V I
authors.initials: VI
authors.lastname: Agol
authors.forename: C U
authors.initials: CU
authors.lastname: Hellen
mesh_terms: D001483:Base Sequence; D018797:Cell Cycle Proteins / Q000502:physiology*; D004247:DNA; D018983:DNA Footprinting; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D014176:Protein Biosynthesis / Q000502:physiology*; D012313:RNA; D017386:Sequence Homology, Amino Acid
chemical_list: D018797:Cell Cycle Proteins; D012313:RNA; D004247:DNA