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A fully orthogonal system for protein synthesis in bacterial cells.
Aleksashin, Nikolay A; Szal, Teresa; d'Aquino, Anne E; Jewett, Michael C; Vázquez-Laslop, Nora; Mankin, Alexander S.
Afiliação
  • Aleksashin NA; Center for Biomolecular Sciences, University of Illinois at Chicago, Chicago, IL, 60607, USA.
  • Szal T; Department of Pharmaceutical Sciences, University of Illinois at Chicago, Chicago, IL, 60612, USA.
  • d'Aquino AE; Center for Biomolecular Sciences, University of Illinois at Chicago, Chicago, IL, 60607, USA.
  • Jewett MC; Interdisciplinary Biological Science Program, Northwestern University, 2145 Sheridan Rd, Evanston, IL, 60208, USA.
  • Vázquez-Laslop N; Department of Chemical and Biological Engineering, Northwestern University, 2145 Sheridan Rd, Evanston, IL, 60208, USA.
  • Mankin AS; Department of Chemical and Biological Engineering, Northwestern University, 2145 Sheridan Rd, Evanston, IL, 60208, USA.
Nat Commun ; 11(1): 1858, 2020 04 20.
Article em En | MEDLINE | ID: mdl-32313034
Ribosome engineering is a powerful approach for expanding the catalytic potential of the protein synthesis apparatus. Due to the potential detriment the properties of the engineered ribosome may have on the cell, the designer ribosome needs to be functionally isolated from the translation machinery synthesizing cellular proteins. One solution to this problem was offered by Ribo-T, an engineered ribosome with tethered subunits which, while producing a desired protein, could be excluded from general translation. Here, we provide a conceptually different design of a cell with two orthogonal protein synthesis systems, where Ribo-T produces the proteome, while the dissociable ribosome is committed to the translation of a specific mRNA. The utility of this system is illustrated by generating a comprehensive collection of mutants with alterations at every rRNA nucleotide of the peptidyl transferase center and isolating gain-of-function variants that enable the ribosome to overcome the translation termination blockage imposed by an arrest peptide.
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Ribossomos / Bactérias / Engenharia de Proteínas / Biologia Sintética Idioma: En Ano de publicação: 2020 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Ribossomos / Bactérias / Engenharia de Proteínas / Biologia Sintética Idioma: En Ano de publicação: 2020 Tipo de documento: Article