Synthetic protein quality control to enhance full-length translation in bacteria
- 4 February 2021
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Chemical Biology
- Vol. 17 (4), 421-427
- https://doi.org/10.1038/s41589-021-00736-3
Abstract
Coupled transcription and translation processes in bacteria cause indiscriminate translation of intact and truncated messenger RNAs, inevitably generating nonfunctional polypeptides. Here, we devised a synthetic protein quality control (ProQC) system that enables translation only when both ends of mRNAs are present and followed by circularization based on sequence-specific RNA–RNA hybridization. We demonstrate that the ProQC system dramatically improved the fraction of full-length proteins among all synthesized polypeptides by selectively translating intact mRNA and reducing abortive translation. As a result, full-length protein synthesis increased up to 2.5-fold without changing the transcription or translation efficiency. Furthermore, we applied the ProQC system for 3-hydroxypropionic acid, violacein and lycopene production by ensuring full-length expression of enzymes in biosynthetic pathways, resulting in 1.6- to 2.3-fold greater biochemical production. We believe that our ProQC system can be universally applied to improve not only the quality of recombinant protein production but also efficiencies of metabolic pathways.Keywords
Funding Information
- National Research Foundation of Korea (2018R1A4A1022513, 2018M3A9H3020459, 2018R1C1B6001129, 2015M3D3A1A01064882)
- Rural Development Administration (PJ01323601)
- Creative-Pioneering Researchers Program through Seoul National University
This publication has 53 references indexed in Scilit:
- Dissection of Malonyl-Coenzyme A Reductase of Chloroflexus aurantiacus Results in Enzyme Activity ImprovementPLOS ONE, 2013
- A Primary Role for Release Factor 3 in Quality Control during Translation Elongation in Escherichia coliCell, 2011
- Condition-Dependent Cell Volume and Concentration of Escherichia coli to Facilitate Data Conversion for Systems Biology ModelingPLOS ONE, 2011
- Protein folding and conformational stress in microbial cells producing recombinant proteins: a host comparative overviewMicrobial Cell Factories, 2008
- Dissociation of halted T7 RNA polymerase elongation complexes proceeds via a forward-translocation mechanismProceedings of the National Academy of Sciences of the United States of America, 2007
- Endonucleolytic cleavage of eukaryotic mRNAs with stalls in translation elongationNature, 2006
- Thinking quantitatively about transcriptional regulationNature Reviews Molecular Cell Biology, 2005
- An mRNA Surveillance Mechanism That Eliminates Transcripts Lacking Termination CodonsScience, 2002
- Exosome-Mediated Recognition and Degradation of mRNAs Lacking a Termination CodonScience, 2002
- Single-Molecule Study of Transcriptional Pausing and Arrest by E. coli RNA PolymeraseScience, 2000