Twice exploration of tRNA +1 frameshifting in an elongation cycle of protein synthesis
| Autor: | Howard Gamper, Yuhong Wang, Henri McGuigan, Gregor Blaha, Yujia Mao, Shoujun Xu, Ya-Ming Hou, Isao Masuda |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Reading Frames
AcademicSubjects/SCI00010 Peptide Chain Elongation Translational RNA Transfer Amino Acyl Biology Ribosome RNA Transfer RNA and RNA-protein complexes Anticodon Escherichia coli Genetics Protein biosynthesis P-site Nucleotide RNA Messenger Codon chemistry.chemical_classification Binding Sites Escherichia coli Proteins Frameshifting Ribosomal Ribosomal RNA Cell biology Amino acid A-site chemistry Transfer RNA Carrier Proteins Ribosomes |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/gkab734 |
| Popis: | Inducing tRNA +1 frameshifting to read a quadruplet codon has the potential to incorporate a non-natural amino acid into the polypeptide chain. While this strategy is being considered for genome expansion in biotechnology and bioengineering endeavors, a major limitation is a lack of understanding of where the shift occurs in an elongation cycle of protein synthesis. Here, we use the high-efficiency +1-frameshifting SufB2 tRNA, containing an extra nucleotide in the anticodon loop, to address this question. Physical and kinetic measurements of the ribosome reading frame of SufB2 identify twice exploration of +1 frameshifting in one elongation cycle, with the major fraction making the shift during translocation from the aminoacyl-tRNA binding (A) site to the peptidyl-tRNA binding (P) site and the remaining fraction making the shift within the P site upon occupancy of the A site in the +1-frame. We demonstrate that the twice exploration of +1 frameshifting occurs during active protein synthesis and that each exploration is consistent with ribosomal conformational dynamics that permits changes of the reading frame. This work indicates that the ribosome itself is a determinant of changes of the reading frame and reveals a mechanistic parallel of +1 frameshifting with –1 frameshifting. Graphical Abstract Graphical AbstracttRNA+1 frameshifting during translocation and in the P site. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|