Remodeling the cellular stress response for enhanced genetic code expansion in mammalian cells.

Autor: Sushkin ME; Biocenter, Johannes Gutenberg University Mainz, Hanns-Dieter-Hüsch-Weg 17, 55128, Mainz, Germany.; International PhD Programme of the Institute of Molecular Biology, Hanns-Dieter-Hüsch-Weg 17, 55128, Mainz, Germany., Koehler C; Biocenter, Johannes Gutenberg University Mainz, Hanns-Dieter-Hüsch-Weg 17, 55128, Mainz, Germany.; VERAXA Biotech GmbH, Carl-Friedrich-Gauß-Ring 5, 69124, Heidelberg, Germany., Lemke EA; Biocenter, Johannes Gutenberg University Mainz, Hanns-Dieter-Hüsch-Weg 17, 55128, Mainz, Germany. edlemke@uni-mainz.de.; Institute of Molecular Biology gGmbH, Ackermannweg 4, 55128, Mainz, Germany. edlemke@uni-mainz.de.
Jazyk: angličtina
Zdroj: Nature communications [Nat Commun] 2023 Oct 31; Vol. 14 (1), pp. 6931. Date of Electronic Publication: 2023 Oct 31.
DOI: 10.1038/s41467-023-42689-2
Abstrakt: Genetic code expansion (GCE) reprograms the translational machinery to site-specifically incorporate noncanonical amino acids (ncAAs) into a selected protein. The efficiency of GCE in mammalian cells might be compromised by cellular stress responses, among which, the protein kinase R(PKR)-dependent eIF2α phosphorylation pathway can reduce translation rates. Here we test several strategies to engineer the eIF2α pathway and boost the rate of translation and show that such interventions increase GCE efficiency in mammalian cells. In particular, addition of the N-terminal PKR fragment (1-174) provides a substantial enhancement in cytoplasmic GCE and also in GCE realized by OTOs (orthogonally translating designer organelles), which built on the principle of 2D phase separation to enable mRNA-selective ncAA incorporation. Our study demonstrates an approach for improving the efficiency of GCE and provides a means by which the power of designer organelles can be further optimized to tune protein translation.
(© 2023. The Author(s).)
Databáze: MEDLINE
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