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Introduction of an AU-rich Element into the 5’ UTR of mRNAs Enhances Protein Expression in Escherichia coli by S1 Protein and Hfq Protein

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Abstract

AU-rich elements in 5’ untranslated region (UTR) are known to increase translation efficiency by recruiting S1 protein that facilitates the assembly of ribosomes. However, AU-rich elements also serve a binding site for Hfq protein, RNase E, etc. To investigate their roles in translation, mRNAs containing either an AU-rich element, originated from sodB 5’-UTR or a non-AU-rich element were constructed. The non-AU-rich elements were designed to retain the thermodynamics of the AUrich element-containing mRNAs to reduce structural effect on translation. The AU-rich element increased mRNA translation and knock-down of S1 protein decreased the translation of AU-rich element-containing mRNAs, confirming the essential role of S1 protein in translation. When their mRNA levels were measured in hfq-deleted cells, those containing a non-AU-rich element and a high AU-content N-terminal coding sequence decreased, representing an auxiliary role of Hfq in translation, specifically in mRNA protection. Interestingly, despite of decreased mRNA level in hfq-deleted cells, protein production was increased, implying the involvement of unknown factors in translation. Consequently, these results suggest that actively translating ribosomes recruited by S1 protein at an AU-rich element stabilize mRNAs from degradation. In the absence of S1 protein, Hfq protein protects mRNAs from degradation. Moreover, AU-rich elements can be used for improved protein production.

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Acknowledgements

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (No. NRF-2018R1A5A1025077), and this research was also supported by the Chung-Ang University Research Grants in 2019.

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Authors and Affiliations

  1. Department of Biomedical Engineering, Chung-Ang University, Seoul, 06974, Korea

    Hyang-Mi Lee, Jun Ren, Woo Young Kim, Phuong Ngoc Lam Vo & Dokyun Na

  2. Department of Life Science, Chung-Ang University, Seoul, 06974, Korea

    Seong-il Eyun

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  1. Hyang-Mi Lee
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Lee, HM., Ren, J., Kim, W.Y. et al. Introduction of an AU-rich Element into the 5’ UTR of mRNAs Enhances Protein Expression in Escherichia coli by S1 Protein and Hfq Protein. Biotechnol Bioproc E 26, 749–757 (2021). https://doi.org/10.1007/s12257-020-0348-3

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  • DOI: https://doi.org/10.1007/s12257-020-0348-3

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