Ribosome Subunit Stapling for Orthogonal Translation in E. coli.
Ribosome Subunit Stapling for Orthogonal Translation in E. coli.
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DOI：
10.1002/ange.201506311
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发表时间：
2015-10-19
期刊：
影响因子：
--
通讯作者：
Chin JW
中科院分区：
文献类型：
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作者：
Fried SD;Schmied WH;Uttamapinant C;Chin JW
正交大核糖体亚基和小核糖体亚基的创建，它们彼此相互作用，但不与内源核糖体亚基相互作用，将通过使大亚基可进化来扩展我们在核糖体中创建新功能的能力。为此，我们合理设计了一种核糖体RNA，通过RNA钉书钉将核糖体亚基共价连接。钉合的核糖体针对正交 mRNA，允许在大亚基中引入突变，从而减少正交翻译，但对细胞生长的影响最小。我们的方法为正交亚基关联提供了一条有前途的途径，这可能使大亚基中关键功能中心（包括肽基转移酶中心）的进化成为可能，用于细胞中非自然聚合物的合成。
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.