title: Recognition of the amber UAG stop codon by release factor RF1.
abstract: We report the crystal structure of a termination complex containing release factor RF1 bound to the 70S ribosome in response to an amber (UAG) codon at 3.6-A resolution. The amber codon is recognized in the 30S subunit-decoding centre directly by conserved elements of domain 2 of RF1, including T186 of the PVT motif. Together with earlier structures, the mechanisms of recognition of all three stop codons by release factors RF1 and RF2 can now be described. Our structure confirms that the backbone amide of Q230 of the universally conserved GGQ motif is positioned to contribute directly to the catalysis of the peptidyl-tRNA hydrolysis reaction through stabilization of the leaving group and/or transition state. We also observe synthetic-negative interactions between mutations in the switch loop of RF1 and in helix 69 of 23S rRNA, revealing that these structural features interact functionally in the termination process. These findings are consistent with our proposal that structural rearrangements of RF1 and RF2 are critical to accurate translation termination.
authors.affiliation: Department of Molecular, Cell and Developmental Biology, Center for Molecular Biology of RNA, UCSC, Santa Cruz, CA, USA.
authors.forename: Andrei
authors.initials: A
authors.lastname: Korostelev
authors.forename: Jianyu
authors.initials: J
authors.lastname: Zhu
authors.forename: Haruichi
authors.initials: H
authors.lastname: Asahara
authors.forename: Harry F
authors.initials: HF
authors.lastname: Noller
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D055162:Biocatalysis; D018388:Codon, Terminator / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D010454:Peptide Termination Factors / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D016963:Thermus thermophilus / Q000737:chemistry* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D018388:Codon, Terminator; D010454:Peptide Termination Factors