How the initiating ribosome copes with ppGpp to translate mRNAs
During host colonization, bacteria use the alarmones (p)ppGpp to reshape their proteome by acting pleiotropically on DNA, RNA, and protein synthesis. Here, we elucidate how the initiating ribosome senses the cellular pool of guanosine nucleotides and regulates the progression towards protein synthes...
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ndltd-PERUUPC-oai-repositorioacademico.upc.edu.pe-10757-6521892020-07-22T05:22:40Z How the initiating ribosome copes with ppGpp to translate mRNAs Vinogradova, Daria S. Zegarra, Victor Maksimova, Elena Nakamoto, Jose Alberto Kasatsky, Pavel Paleskava, Alena Konevega, Andrey L. Milón, Pohl During host colonization, bacteria use the alarmones (p)ppGpp to reshape their proteome by acting pleiotropically on DNA, RNA, and protein synthesis. Here, we elucidate how the initiating ribosome senses the cellular pool of guanosine nucleotides and regulates the progression towards protein synthesis. Our results show that the affinity of guanosine triphosphate (GTP) and the inhibitory concentration of ppGpp for the 30S-bound initiation factor IF2 vary depending on the programmed mRNA. The TufA mRNA enhanced GTP affinity for 30S complexes, resulting in improved ppGpp tolerance and allowing efficient protein synthesis. Conversely, the InfA mRNA allowed ppGpp to compete with GTP for IF2, thus stalling 30S complexes. Structural modeling and biochemical analysis of the TufA mRNA unveiled a structured enhancer of translation initiation (SETI) composed of two consecutive hairpins proximal to the translation initiation region (TIR) that largely account for ppGpp tolerance under physiological concentrations of guanosine nucleotides. Furthermore, our results show that the mechanism enhancing ppGpp tolerance is not restricted to the TufA mRNA, as similar ppGpp tolerance was found for the SETI-containing Rnr mRNA. Finally, we show that IF2 can use pppGpp to promote the formation of 30S initiation complexes (ICs), albeit requiring higher factor concentration and resulting in slower transitions to translation elongation. Altogether, our data unveil a novel regulatory mechanism at the onset of protein synthesis that tolerates physiological concentrations of ppGpp and that bacteria can exploit to modulate their proteome as a function of the nutritional shift happening during stringent response and infection. Russian Foundation for Basic Research Revisión por pares 2020-07-20T02:05:54Z 2020-07-20T02:05:54Z 2020-01-01 info:eu-repo/semantics/article 15449173 10.1371/journal.pbio.3000593 http://hdl.handle.net/10757/652189 15457885 PLoS Biology 2-s2.0-85078947441 SCOPUS_ID:85078947441 0000 0001 2196 144X eng https://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.3000593 info:eu-repo/semantics/openAccess Attribution-NonCommercial-ShareAlike 4.0 International http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf Public Library of Science Universidad Peruana de Ciencias Aplicadas (UPC) Repositorio Academico - UPC PLoS Biology 18 1 |
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English |
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Article |
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During host colonization, bacteria use the alarmones (p)ppGpp to reshape their proteome by acting pleiotropically on DNA, RNA, and protein synthesis. Here, we elucidate how the initiating ribosome senses the cellular pool of guanosine nucleotides and regulates the progression towards protein synthesis. Our results show that the affinity of guanosine triphosphate (GTP) and the inhibitory concentration of ppGpp for the 30S-bound initiation factor IF2 vary depending on the programmed mRNA. The TufA mRNA enhanced GTP affinity for 30S complexes, resulting in improved ppGpp tolerance and allowing efficient protein synthesis. Conversely, the InfA mRNA allowed ppGpp to compete with GTP for IF2, thus stalling 30S complexes. Structural modeling and biochemical analysis of the TufA mRNA unveiled a structured enhancer of translation initiation (SETI) composed of two consecutive hairpins proximal to the translation initiation region (TIR) that largely account for ppGpp tolerance under physiological concentrations of guanosine nucleotides. Furthermore, our results show that the mechanism enhancing ppGpp tolerance is not restricted to the TufA mRNA, as similar ppGpp tolerance was found for the SETI-containing Rnr mRNA. Finally, we show that IF2 can use pppGpp to promote the formation of 30S initiation complexes (ICs), albeit requiring higher factor concentration and resulting in slower transitions to translation elongation. Altogether, our data unveil a novel regulatory mechanism at the onset of protein synthesis that tolerates physiological concentrations of ppGpp and that bacteria can exploit to modulate their proteome as a function of the nutritional shift happening during stringent response and infection. === Russian Foundation for Basic Research === Revisión por pares |
author |
Vinogradova, Daria S. Zegarra, Victor Maksimova, Elena Nakamoto, Jose Alberto Kasatsky, Pavel Paleskava, Alena Konevega, Andrey L. Milón, Pohl |
spellingShingle |
Vinogradova, Daria S. Zegarra, Victor Maksimova, Elena Nakamoto, Jose Alberto Kasatsky, Pavel Paleskava, Alena Konevega, Andrey L. Milón, Pohl How the initiating ribosome copes with ppGpp to translate mRNAs |
author_facet |
Vinogradova, Daria S. Zegarra, Victor Maksimova, Elena Nakamoto, Jose Alberto Kasatsky, Pavel Paleskava, Alena Konevega, Andrey L. Milón, Pohl |
author_sort |
Vinogradova, Daria S. |
title |
How the initiating ribosome copes with ppGpp to translate mRNAs |
title_short |
How the initiating ribosome copes with ppGpp to translate mRNAs |
title_full |
How the initiating ribosome copes with ppGpp to translate mRNAs |
title_fullStr |
How the initiating ribosome copes with ppGpp to translate mRNAs |
title_full_unstemmed |
How the initiating ribosome copes with ppGpp to translate mRNAs |
title_sort |
how the initiating ribosome copes with ppgpp to translate mrnas |
publisher |
Public Library of Science |
publishDate |
2020 |
url |
http://hdl.handle.net/10757/652189 |
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