Three-dimensional structure-guided evolution of a ribosome with tethered subunits
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- Title
- Three-dimensional structure-guided evolution of a ribosome with tethered subunits
- Authors
- Kim, Do Soon; Watkins, Andrew; Bidstrup, Erik; Lee, Joongoo; Topkar, Ved; Kofman, Camila; Schwarz, Kevin J.; Liu, Yan; Pintilie, Grigore; Roney, Emily; Das, Rhiju; Jewett, Michael C.
- Date Issued
- 2022-09
- Publisher
- Nature Publishing Group
- 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 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. [Figure not available: see fulltext.] © 2022, The Author(s), under exclusive licence to Springer Nature America, Inc.
- URI
- https://oasis.postech.ac.kr/handle/2014.oak/116188
- DOI
- 10.1038/s41589-022-01064-w
- ISSN
- 1552-4450
- Article Type
- Article
- Citation
- Nature Chemical Biology, vol. 18, no. 9, page. 990 - 998, 2022-09
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