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      Increased mistranslation protects E. coli from protein misfolding stress due to activation of a RpoS‐dependent heat shock response

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          Abstract

          The misincorporation of an incorrect amino acid into a polypeptide during protein synthesis is considered a detrimental phenomenon. A mistranslated protein is often misfolded and degraded or nonfunctional and results in an increased cost to quality control machinery. Despite these costs, errors during protein synthesis are common in bacteria. Here, we report that mistranslation in Escherichia coli increase the protein level of the heat shock sigma factor RpoH and protect cells against heat stress. Surprisingly, this increase in RpoH due to mistranslation is dependent on the presence of the general stress response sigma factor RpoS. This report provides evidence for a protective function of mistranslation and suggests a novel regulatory role of RpoS in the heat shock response.

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          Most cited references27

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          The heat shock response: life on the verge of death.

          Organisms must survive a variety of stressful conditions, including sudden temperature increases that damage important cellular structures and interfere with essential functions. In response to heat stress, cells activate an ancient signaling pathway leading to the transient expression of heat shock or heat stress proteins (Hsps). Hsps exhibit sophisticated protection mechanisms, and the most conserved Hsps are molecular chaperones that prevent the formation of nonspecific protein aggregates and assist proteins in the acquisition of their native structures. In this Review, we summarize the concepts of the protective Hsp network. Copyright © 2010 Elsevier Inc. All rights reserved.
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            Mistranslation of membrane proteins and two-component system activation trigger antibiotic-mediated cell death.

            Aminoglycoside antibiotics, such as gentamicin and kanamycin, directly target the ribosome, yet the mechanisms by which these bactericidal drugs induce cell death are not fully understood. Recently, oxidative stress has been implicated as one of the mechanisms whereby bactericidal antibiotics kill bacteria. Here, we use systems-level approaches and phenotypic analyses to provide insight into the pathway whereby aminoglycosides ultimately trigger hydroxyl radical formation. We show, by disabling systems that facilitate membrane protein traffic, that mistranslation and misfolding of membrane proteins are central to aminoglycoside-induced oxidative stress and cell death. Signaling through the envelope stress-response two-component system is found to be a key player in this process, and the redox-responsive two-component system is shown to have an associated role. Additionally, we show that these two-component systems play a general role in bactericidal antibiotic-mediated oxidative stress and cell death, expanding our understanding of the common mechanism of killing induced by bactericidal antibiotics.
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              Innate Immune and Chemically Triggered Oxidative Stress Modifies Translational Fidelity

              Translational fidelity, essential for protein and cell function, requires accurate tRNA aminoacylation. Purified aminoacyl-tRNA synthetases exhibit a fidelity of 1 error per 10,000 to 100,000 couplings 1, 2. The accuracy of tRNA aminoacylation in vivo is uncertain, however, and might be considerably lower 3–6. Here, we show that in mammalian cells, approximately 1% of methionine (Met) residues used in protein synthesis are aminoacylated to non-methionyl-tRNAs. Remarkably, Met-misacylation increases up to 10-fold upon exposing cells to live or non-infectious viruses, toll-like receptor ligands, or chemically induced oxidative stress. Met is misacylated to specific non-methionyl-tRNA families, and these Met-misacylated tRNAs are used in translation. Met-misacylation is blocked by an inhibitor of cellular oxidases, implicating reactive oxygen species (ROS) as the misacylation trigger. Among six amino acids tested, tRNA misacylation occurs exclusively with Met. As Met residues are known to protect proteins against ROS-mediated damage 7, we propose that Met-misacylation functions adaptively to increase Met incorporation into proteins to protect cells against oxidative stress. In demonstrating an unexpected conditional aspect of decoding mRNA, our findings illustrate the importance of considering alternative iterations of the genetic code.
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                Author and article information

                Contributors
                jling12@umd.edu
                Journal
                FEBS Lett
                FEBS Lett
                10.1002/(ISSN)1873-3468
                FEB2
                Febs Letters
                John Wiley and Sons Inc. (Hoboken )
                0014-5793
                1873-3468
                24 August 2019
                November 2019
                : 593
                : 22 ( doiID: 10.1002/feb2.v593.22 )
                : 3220-3227
                Affiliations
                [ 1 ] Department of Microbiology and Molecular Genetics McGovern Medical School University of Texas Health Science Center Houston TX USA
                [ 2 ]Present address: Department of Biological Sciences Columbia University New York NY 10027 USA
                [ 3 ]Present address: College of Life and Health Sciences Northeastern University Shenyang 110819 China
                Author notes
                [*] [* ] Correspondence

                J. Ling, Department of Cell Biology and Molecular Genetics, The University of Maryland, College Park, MD 20742, USA

                Tel: +1 301 405 1035

                E‐mail: jling12@ 123456umd.edu

                Author information
                https://orcid.org/0000-0003-4466-8304
                Article
                FEB213578
                10.1002/1873-3468.13578
                6878130
                31419308
                e6651d5c-5e27-4c42-99e6-64fd6f071adf
                © 2019 The Authors. FEBS Letters published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.

                History
                : 10 July 2019
                : 02 August 2019
                : 08 August 2019
                Page count
                Figures: 5, Tables: 0, Pages: 8, Words: 4618
                Funding
                Funded by: National Institute of General Medical Sciences , open-funder-registry 10.13039/100000057;
                Award ID: R01GM115431
                Funded by: National Institute of Neurological Disorders and Stroke , open-funder-registry 10.13039/100000065;
                Award ID: R21NS101245
                Categories
                Research Letter
                Research Letters
                Microbiology
                Custom metadata
                2.0
                November 2019
                Converter:WILEY_ML3GV2_TO_JATSPMC version:5.7.2 mode:remove_FC converted:05.12.2019

                Molecular biology
                heat shock response,mistranslation,rpoh,rpos
                Molecular biology
                heat shock response, mistranslation, rpoh, rpos

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