6HRM

E. coli 70S d2d8 stapled ribosome

これはPDB形式変換不可エントリーです。

6HRM の概要

• エントリーDOI: 10.2210/pdb6hrm/pdb
• EMDBエントリー: 0261
• 分子名称: stapled 16S-23S rRNA,stapled 16S-23S rRNA,stapled 16S-23S rRNA,stapled 16S-23S rRNA, 50S ribosomal protein L11, 50S ribosomal protein L13, ... (56 entities in total)
• 機能のキーワード: staple, ribosome
• 由来する生物種: Escherichia coli; 詳細
• タンパク質・核酸の鎖数: 53
• 化学式量合計: 2159788.00

構造登録者

Schmied, W.H.,Tnimov, Z.,Uttamapinant, C.,Rae, C.D.,Fried, S.D.,Chin, J.W. (登録日: 2018-09-27, 公開日: 2018-12-19, 最終更新日: 2025-03-12)

主引用文献

Schmied, W.H.,Tnimov, Z.,Uttamapinant, C.,Rae, C.D.,Fried, S.D.,Chin, J.W.
Controlling orthogonal ribosome subunit interactions enables evolution of new function.
Nature, 564:444-448, 2018

PubMed Abstract: Orthogonal ribosomes are unnatural ribosomes that are directed towards orthogonal messenger RNAs in Escherichia coli, through an altered version of the 16S ribosomal RNA of the small subunit. Directed evolution of orthogonal ribosomes has provided access to new ribosomal function, and the evolved orthogonal ribosomes have enabled the encoding of multiple non-canonical amino acids into proteins. The original orthogonal ribosomes shared the pool of 23S ribosomal RNAs, contained in the large subunit, with endogenous ribosomes. Selectively directing a new 23S rRNA to an orthogonal mRNA, by controlling the association between the orthogonal 16S rRNAs and 23S rRNAs, would enable the evolution of new function in the large subunit. Previous work covalently linked orthogonal 16S rRNA and a circularly permuted 23S rRNA to create orthogonal ribosomes with low activity; however, the linked subunits in these ribosomes do not associate specifically with each other, and mediate translation by associating with endogenous subunits. Here we discover engineered orthogonal 'stapled' ribosomes (with subunits linked through an optimized RNA staple) with activities comparable to that of the parent orthogonal ribosome; they minimize association with endogenous subunits and mediate translation of orthogonal mRNAs through the association of stapled subunits. We evolve cells with genomically encoded stapled ribosomes as the sole ribosomes, which support cellular growth at similar rates to natural ribosomes. Moreover, we visualize the engineered stapled ribosome structure by cryo-electron microscopy at 3.0 Å, revealing how the staple links the subunits and controls their association. We demonstrate the utility of controlling subunit association by evolving orthogonal stapled ribosomes which efficiently polymerize a sequence of monomers that the natural ribosome is intrinsically unable to translate. Our work provides a foundation for evolving the rRNA of the entire orthogonal ribosome for the encoded cellular synthesis of non-canonical biological polymers.

PubMed: 30518861
DOI: 10.1038/s41586-018-0773-z

実験手法

ELECTRON MICROSCOPY (2.96 Å)