Journal
SCIENCE ADVANCES
Volume 9, Issue 17, Pages -Publisher
AMER ASSOC ADVANCEMENT SCIENCE
DOI: 10.1126/sciadv.ade8934
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This study develops a method to map the nucleotides in the peptidyl transfer center and the resulting epistatic interactions.
Fitness landscapes are models of the sequence space of a genetic element that map how each sequence corre-sponds to its activity and can be used to guide laboratory evolution. The ribosome is a macromolecular machine that is essential for protein synthesis in all organisms. Because of the prevalence of dominant lethal mutations, a comprehensive fitness landscape of the ribosomal peptidyl transfer center (PTC) has not yet been attained. Here, we develop a method to functionally map an orthogonal tethered ribosome (oRiboT), which permits com-plete mutagenesis of nucleotides located in the PTC and the resulting epistatic interactions. We found that most nucleotides studied showed flexibility to mutation, and identified epistatic interactions between them, which compensate for deleterious mutations. This work provides a basis for a deeper understanding of ribosome func-tion and malleability and could be used to inform design of engineered ribosomes with applications to synthe-size next-generation biomaterials and therapeutics.
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