title: In vitro suppression as a tool for the investigation of translation initiation.
abstract: An in vitro protein synthesizing system that employs rabbit reticulocyte lysates has been employed for protein production from mRNAs containing nonsense (UAG) codons in the presence of misacylated suppressor tRNAs.The system includes a misacylated Escherichia coli tRNAAlaCUA that functions at least as efficiently as any suppressor tRNA transcript reported to date and which has been shown not to be a substrate for (re)activation by alanyl-tRNA synthetase. Application of the optimized system for preparation of dihydrofolate analogs has also permitted analysis of competing mechanisms that control the sites(s) of translation initiation.
authors.affiliation: Departments of Chemistry and Biology, University of Virginia, Charlottesville, VA 22901, USA.
authors.forename: V A
authors.initials: VA
authors.lastname: Karginov
authors.forename: S V
authors.initials: SV
authors.lastname: Mamaev
authors.forename: S M
authors.initials: SM
authors.lastname: Hecht
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018389:Codon, Nonsense / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D010442:Peptide Chain Initiation, Translational / Q000235:genetics*; D012329:RNA, Bacterial / Q000235:genetics / Q000378:metabolism; D012344:RNA, Transfer, Ala / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism; D013489:Suppression, Genetic; D013762:Tetrahydrofolate Dehydrogenase / Q000096:biosynthesis / Q000235:genetics
chemical_list: D018389:Codon, Nonsense; D012329:RNA, Bacterial; D012344:RNA, Transfer, Ala; D013762:Tetrahydrofolate Dehydrogenase