Translation initiation with initiator tRNA charged with exotic peptides.
In the prokaryotic translation system formylmethionyl-tRNA(fMet)(CAU) acts as an exclusive initiator to yield peptides bearing formylmethionine at the N-terminus. We herein report a new methodology to initiate the translation reaction with peptidyl-tRNA(fMet)(CAU) in which the peptidyl group consists of unusual building blocks, such as D-amino acids, beta-amino acids, and N-methyl amino acids, and express peptides containing a stretch of exotic peptides at the N-terminus. Synthesis of the initiator peptidyl-tRNAs was facilitated by flexizyme, a flexible tRNA aminoacylation ribozyme, and the start codon was reprogrammed by withdrawing methionine from a reconstituted E. coli cell-free translation system to reassign the initiator from formylmethionine to exotic peptides. This represents the first demonstration for initiating the ribosomal peptide synthesis with peptides, which offers us a new tool for the synthesis of a wide variety of unusual peptides and also the mechanistic studies on the initiation and/or elongation events in translation.
Goto
Yuki
Y
Research Center for Advanced Science and Technology and Department of Advanced Interdisciplinary Studies, The University of Tokyo, 4-6-1, Komaba, Meguro-ku, Tokyo 153-8904, Japan.
Suga
Hiroaki
H
D000596:Amino Acids / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics; D010442:Peptide Chain Initiation, Translational; D010455:Peptides / Q000737:chemistry* / Q000235:genetics*; D012329:RNA, Bacterial / Q000737:chemistry / Q000235:genetics; D012346:RNA, Transfer, Amino Acyl / Q000737:chemistry* / Q000235:genetics*; D012358:RNA, Transfer, Met / Q000737:chemistry / Q000235:genetics; D012270:Ribosomes / Q000235:genetics; D046249:Transfer RNA Aminoacylation
D000596:Amino Acids; D010455:Peptides; D012329:RNA, Bacterial; D012346:RNA, Transfer, Amino Acyl; D012358:RNA, Transfer, Met; C019037:tRNA, peptidyl-