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Ribosome Subunit Stapling for Orthogonal Translation in E. coli

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Specialty Chemistry
Date 2015 Oct 15
PMID 26465656
Citations 34
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Abstract

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.

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References
1.
Szewczak A, Cech T . An RNA internal loop acts as a hinge to facilitate ribozyme folding and catalysis. RNA. 1997; 3(8):838-49. PMC: 1369529. View

2.
Yokoyama T, Suzuki T . Ribosomal RNAs are tolerant toward genetic insertions: evolutionary origin of the expansion segments. Nucleic Acids Res. 2008; 36(11):3539-51. PMC: 2441807. DOI: 10.1093/nar/gkn224. View

3.
Aitken C, Puglisi J . Following the intersubunit conformation of the ribosome during translation in real time. Nat Struct Mol Biol. 2010; 17(7):793-800. PMC: 4459212. DOI: 10.1038/nsmb.1828. View

4.
Schmeing T, Ramakrishnan V . What recent ribosome structures have revealed about the mechanism of translation. Nature. 2009; 461(7268):1234-42. DOI: 10.1038/nature08403. View

5.
Triman K . Mutational analysis of the ribosome. Adv Genet. 2007; 58:89-119. DOI: 10.1016/S0065-2660(06)58004-6. View