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Ribosome Subunit Stapling for Orthogonal Translation in E. coli
Author(s) -
Fried Stephen D.,
Schmied Wolfgang H.,
Uttamapinant Chayasith,
Chin Jason W.
Publication year - 2015
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201506311
Subject(s) - translation (biology) , ribosome , protein subunit , chemistry , computational biology , stereochemistry , biochemistry , biology , rna , messenger rna , gene
The creation of orthogonal large and small ribosomal subunits, which interact with each other but not with endogenous ribosomal subunits, would extend our capacity to create new functions in the ribosome by making the large subunit evolvable. To this end, we rationally designed a ribosomal RNA that covalently links the ribosome subunits via an RNA staple. The stapled ribosome is directed to an orthogonal mRNA, allowing the introduction of mutations into the large subunit that reduce orthogonal translation, but have minimal effects on cell growth. Our approach provides a promising route towards orthogonal subunit association, which may enable the evolution of key functional centers in the large subunit, including the peptidyl‐transferase center, for unnatural polymer synthesis in cells.