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TitleThree-dimensional structure-guided evolution of a ribosome with tethered subunits.
Journal, issue, pagesNat Chem Biol, Vol. 18, Issue 9, Page 990-998, Year 2022
Publish dateJul 14, 2022
AuthorsDo 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 AbstractRNA-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 linksNat Chem Biol / PubMed:35836020 / PubMed Central
MethodsEM (single particle)
Resolution4.18 Å
Structure data

26666

7uph

EMDB-26666, PDB-7uph:
Structure of a ribosome with tethered subunits
Method: EM (single particle) / Resolution: 4.18 Å

Source
  • escherichia coli (E. coli)
  • escherichia coli k-12 (bacteria)
  • Escherichia coli
KeywordsRIBOSOME / Engineered / Tethered / Synthetic

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