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-Structure paper
Title | Three-dimensional structure-guided evolution of a ribosome with tethered subunits. |
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Journal, issue, pages | Nat Chem Biol, Vol. 18, Issue 9, Page 990-998, Year 2022 |
Publish date | Jul 14, 2022 |
Authors | Do Soon Kim / Andrew Watkins / Erik Bidstrup / Joongoo Lee / Ved Topkar / Camila Kofman / Kevin J Schwarz / Yan Liu / Grigore Pintilie / Emily Roney / Rhiju Das / Michael C Jewett / |
PubMed Abstract | RNA-based macromolecular machines, such as the ribosome, have functional parts reliant on structural interactions spanning sequence-distant regions. These features limit evolutionary exploration of ...RNA-based macromolecular machines, such as the ribosome, have functional parts reliant on structural interactions spanning sequence-distant regions. These features limit evolutionary exploration of mutant libraries and confound three-dimensional structure-guided design. To address these challenges, we describe Evolink (evolution and linkage), a method that enables high-throughput evolution of sequence-distant regions in large macromolecular machines, and library design guided by computational RNA modeling to enable exploration of structurally stable designs. Using Evolink, we evolved a tethered ribosome with a 58% increased activity in orthogonal protein translation and a 97% improvement in doubling times in SQ171 cells compared to a previously developed tethered ribosome, and reveal new permissible sequences in a pair of ribosomal helices with previously explored biological function. The Evolink approach may enable enhanced engineering of macromolecular machines for new and improved functions for synthetic biology. |
External links | Nat Chem Biol / PubMed:35836020 / PubMed Central |
Methods | EM (single particle) |
Resolution | 4.18 Å |
Structure data | EMDB-26666, PDB-7uph: |
Source |
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Keywords | RIBOSOME / Engineered / Tethered / Synthetic |