title: Efficient 50S ribosome-catalyzed peptide bond synthesis with an aminoacyl minihelix.
abstract: RNA minihelices that recreate the amino acid acceptor domain of the two-domain L-shaped tRNA molecule are substrates for specific aminoacylation by tRNA synthetases. Some lines of evidence suggest that this domain arose independently of and predated the second, anticodon-containing domain. With puromycin and a minihelix charged with alanine, we show here efficient 50S ribosome catalyzed peptide synthesis. The aminoacyl minihelix is as active as aminoacyl tRNA in the synthetic reaction. The high efficiency of the charged minihelix is due to a relatively strong interaction with the 50S particle. In contrast, an aminoacyl RNA fragment that recreates the 3'-side of the tRNA acceptor stem has a much weaker interaction with the 50S particle. These results are consistent with the minihelix domain being the major loci for tRNA interactions with the 50S ribosome. They may also have implications for the historical development of RNA-based systems of peptide synthesis.
authors.affiliation: The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
authors.forename: N Y
authors.initials: NY
authors.lastname: Sardesai
authors.forename: R
authors.initials: R
authors.lastname: Green
authors.forename: P
authors.initials: P
authors.lastname: Schimmel
mesh_terms: D000215:Acylation; D000412:Alanine-tRNA Ligase / Q000378:metabolism; D002384:Catalysis; D002855:Chromatography, Thin Layer; D009690:Nucleic Acid Conformation; D009842:Oligopeptides / Q000378:metabolism; D010452:Peptide Biosynthesis; D010458:Peptidyl Transferases / Q000378:metabolism; D011691:Puromycin / Q000031:analogs & derivatives / Q000096:biosynthesis; D012335:RNA, Ribosomal / Q000737:chemistry / Q000378:metabolism*; D012344:RNA, Transfer, Ala / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity; D046249:Transfer RNA Aminoacylation
chemical_list: D009842:Oligopeptides; D012335:RNA, Ribosomal; D012344:RNA, Transfer, Ala; C046712:N-acetylphenylalanylpuromycin; D011691:Puromycin; D010458:Peptidyl Transferases; D000412:Alanine-tRNA Ligase