Multifaceted Mechanism of Amicoumacin A Inhibition of Bacterial Translation
Amicoumacin A (Ami) halts bacterial growth by inhibiting the ribosome during translation. The Ami binding site locates in the vicinity of the E-site codon of mRNA. However, Ami does not clash with mRNA, rather stabilizes it, which is relatively unusual and implies a unique way of translation inhibit...
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ndltd-PERUUPC-oai-repositorioacademico.upc.edu.pe-10757-6558222021-05-06T05:11:34Z Multifaceted Mechanism of Amicoumacin A Inhibition of Bacterial Translation Maksimova, Elena M. Vinogradova, Daria S. Osterman, Ilya A. Kasatsky, Pavel S. Nikonov, Oleg S. Milón, Pohl Dontsova, Olga A. Sergiev, Petr V. Paleskava, Alena Konevega, Andrey L. amicoumacin A antibiotic resistance elongation factor EF-G initiation microscale thermophoresis rapid kinetics translocation Amicoumacin A (Ami) halts bacterial growth by inhibiting the ribosome during translation. The Ami binding site locates in the vicinity of the E-site codon of mRNA. However, Ami does not clash with mRNA, rather stabilizes it, which is relatively unusual and implies a unique way of translation inhibition. In this work, we performed a kinetic and thermodynamic investigation of Ami influence on the main steps of polypeptide synthesis. We show that Ami reduces the rate of the functional canonical 70S initiation complex (IC) formation by 30-fold. Additionally, our results indicate that Ami promotes the formation of erroneous 30S ICs; however, IF3 prevents them from progressing towards translation initiation. During early elongation steps, Ami does not compromise EF-Tu-dependent A-site binding or peptide bond formation. On the other hand, Ami reduces the rate of peptidyl-tRNA movement from the A to the P site and significantly decreases the amount of the ribosomes capable of polypeptide synthesis. Our data indicate that Ami progressively decreases the activity of translating ribosomes that may appear to be the main inhibitory mechanism of Ami. Indeed, the use of EF-G mutants that confer resistance to Ami (G542V, G581A, or ins544V) leads to a complete restoration of the ribosome functionality. It is possible that the changes in translocation induced by EF-G mutants compensate for the activity loss caused by Ami. Russian Foundation for Basic Research Revisión por pares 2021-05-04T16:06:57Z 2021-05-04T16:06:57Z 2021-02-12 info:eu-repo/semantics/article 10.3389/fmicb.2021.618857 http://hdl.handle.net/10757/655822 1664302X Frontiers in Microbiology 2-s2.0-85101886894 SCOPUS_ID:85101886894 0000 0001 2196 144X eng https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7907450/ info:eu-repo/semantics/openAccess Attribution-NonCommercial-ShareAlike 4.0 International http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf Frontiers Media S.A. Universidad Peruana de Ciencias Aplicadas (UPC) Repositorio Academico - UPC Frontiers in Microbiology 12 |
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English |
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Article |
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amicoumacin A antibiotic resistance elongation factor EF-G initiation microscale thermophoresis rapid kinetics translocation |
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amicoumacin A antibiotic resistance elongation factor EF-G initiation microscale thermophoresis rapid kinetics translocation Maksimova, Elena M. Vinogradova, Daria S. Osterman, Ilya A. Kasatsky, Pavel S. Nikonov, Oleg S. Milón, Pohl Dontsova, Olga A. Sergiev, Petr V. Paleskava, Alena Konevega, Andrey L. Multifaceted Mechanism of Amicoumacin A Inhibition of Bacterial Translation |
description |
Amicoumacin A (Ami) halts bacterial growth by inhibiting the ribosome during translation. The Ami binding site locates in the vicinity of the E-site codon of mRNA. However, Ami does not clash with mRNA, rather stabilizes it, which is relatively unusual and implies a unique way of translation inhibition. In this work, we performed a kinetic and thermodynamic investigation of Ami influence on the main steps of polypeptide synthesis. We show that Ami reduces the rate of the functional canonical 70S initiation complex (IC) formation by 30-fold. Additionally, our results indicate that Ami promotes the formation of erroneous 30S ICs; however, IF3 prevents them from progressing towards translation initiation. During early elongation steps, Ami does not compromise EF-Tu-dependent A-site binding or peptide bond formation. On the other hand, Ami reduces the rate of peptidyl-tRNA movement from the A to the P site and significantly decreases the amount of the ribosomes capable of polypeptide synthesis. Our data indicate that Ami progressively decreases the activity of translating ribosomes that may appear to be the main inhibitory mechanism of Ami. Indeed, the use of EF-G mutants that confer resistance to Ami (G542V, G581A, or ins544V) leads to a complete restoration of the ribosome functionality. It is possible that the changes in translocation induced by EF-G mutants compensate for the activity loss caused by Ami. === Russian Foundation for Basic Research === Revisión por pares |
author |
Maksimova, Elena M. Vinogradova, Daria S. Osterman, Ilya A. Kasatsky, Pavel S. Nikonov, Oleg S. Milón, Pohl Dontsova, Olga A. Sergiev, Petr V. Paleskava, Alena Konevega, Andrey L. |
author_facet |
Maksimova, Elena M. Vinogradova, Daria S. Osterman, Ilya A. Kasatsky, Pavel S. Nikonov, Oleg S. Milón, Pohl Dontsova, Olga A. Sergiev, Petr V. Paleskava, Alena Konevega, Andrey L. |
author_sort |
Maksimova, Elena M. |
title |
Multifaceted Mechanism of Amicoumacin A Inhibition of Bacterial Translation |
title_short |
Multifaceted Mechanism of Amicoumacin A Inhibition of Bacterial Translation |
title_full |
Multifaceted Mechanism of Amicoumacin A Inhibition of Bacterial Translation |
title_fullStr |
Multifaceted Mechanism of Amicoumacin A Inhibition of Bacterial Translation |
title_full_unstemmed |
Multifaceted Mechanism of Amicoumacin A Inhibition of Bacterial Translation |
title_sort |
multifaceted mechanism of amicoumacin a inhibition of bacterial translation |
publisher |
Frontiers Media S.A. |
publishDate |
2021 |
url |
http://hdl.handle.net/10757/655822 |
work_keys_str_mv |
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