Structural highlights
FunctionRL3_ECOLI One of two assembly inititator proteins, it binds directly near the 3'-end of the 23S rRNA, where it nucleates assembly of the 50S subunit.[HAMAP-Rule:MF_01325_B] Publication Abstract from PubMedRNA-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. Three-dimensional structure-guided evolution of a ribosome with tethered subunits.,Kim DS, Watkins A, Bidstrup E, Lee J, Topkar V, Kofman C, Schwarz KJ, Liu Y, Pintilie G, Roney E, Das R, Jewett MC Nat Chem Biol. 2022 Sep;18(9):990-998. doi: 10.1038/s41589-022-01064-w. Epub 2022 , Jul 14. PMID:35836020[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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