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Regulation of Ribosomal Protein Synthesis in Mycobacteria: The Autogenous Control of rpsO
The autogenous regulation of ribosomal protein (r-protein) synthesis plays a key role in maintaining the stoichiometry of ribosomal components in bacteria. In this work, taking the rpsO gene as a classic example, we addressed for the first time the in vivo regulation of r-protein synthesis in the my...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8470358/ https://www.ncbi.nlm.nih.gov/pubmed/34575857 http://dx.doi.org/10.3390/ijms22189679 |
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author | Aseev, Leonid V. Koledinskaya, Ludmila S. Bychenko, Oksana S. Boni, Irina V. |
author_facet | Aseev, Leonid V. Koledinskaya, Ludmila S. Bychenko, Oksana S. Boni, Irina V. |
author_sort | Aseev, Leonid V. |
collection | PubMed |
description | The autogenous regulation of ribosomal protein (r-protein) synthesis plays a key role in maintaining the stoichiometry of ribosomal components in bacteria. In this work, taking the rpsO gene as a classic example, we addressed for the first time the in vivo regulation of r-protein synthesis in the mycobacteria M. smegmatis (Msm) and M. tuberculosis (Mtb). We used a strategy based on chromosomally integrated reporters under the control of the rpsO regulatory regions and the ectopic expression of Msm S15 to measure its impact on the reporter expression. Because the use of E. coli as a host appeared inefficient, a fluorescent reporter system was developed by inserting Msm or Mtb rpsO-egfp fusions into the Msm chromosome and expressing Msm S15 or E. coli S15 in trans from a novel replicative shuttle vector, pAMYC. The results of the eGFP expression measurements in Msm cells provided evidence that the rpsO gene in Msm and Mtb was feedback-regulated at the translation level. The mutagenic analysis showed that the folding of Msm rpsO 5′UTR in a pseudoknot appeared crucial for repression by both Msm S15 and E. coli S15, thus indicating a striking resemblance of the rpsO feedback control in mycobacteria and in E. coli. |
format | Online Article Text |
id | pubmed-8470358 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-84703582021-09-27 Regulation of Ribosomal Protein Synthesis in Mycobacteria: The Autogenous Control of rpsO Aseev, Leonid V. Koledinskaya, Ludmila S. Bychenko, Oksana S. Boni, Irina V. Int J Mol Sci Article The autogenous regulation of ribosomal protein (r-protein) synthesis plays a key role in maintaining the stoichiometry of ribosomal components in bacteria. In this work, taking the rpsO gene as a classic example, we addressed for the first time the in vivo regulation of r-protein synthesis in the mycobacteria M. smegmatis (Msm) and M. tuberculosis (Mtb). We used a strategy based on chromosomally integrated reporters under the control of the rpsO regulatory regions and the ectopic expression of Msm S15 to measure its impact on the reporter expression. Because the use of E. coli as a host appeared inefficient, a fluorescent reporter system was developed by inserting Msm or Mtb rpsO-egfp fusions into the Msm chromosome and expressing Msm S15 or E. coli S15 in trans from a novel replicative shuttle vector, pAMYC. The results of the eGFP expression measurements in Msm cells provided evidence that the rpsO gene in Msm and Mtb was feedback-regulated at the translation level. The mutagenic analysis showed that the folding of Msm rpsO 5′UTR in a pseudoknot appeared crucial for repression by both Msm S15 and E. coli S15, thus indicating a striking resemblance of the rpsO feedback control in mycobacteria and in E. coli. MDPI 2021-09-07 /pmc/articles/PMC8470358/ /pubmed/34575857 http://dx.doi.org/10.3390/ijms22189679 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Aseev, Leonid V. Koledinskaya, Ludmila S. Bychenko, Oksana S. Boni, Irina V. Regulation of Ribosomal Protein Synthesis in Mycobacteria: The Autogenous Control of rpsO |
title | Regulation of Ribosomal Protein Synthesis in Mycobacteria: The Autogenous Control of rpsO |
title_full | Regulation of Ribosomal Protein Synthesis in Mycobacteria: The Autogenous Control of rpsO |
title_fullStr | Regulation of Ribosomal Protein Synthesis in Mycobacteria: The Autogenous Control of rpsO |
title_full_unstemmed | Regulation of Ribosomal Protein Synthesis in Mycobacteria: The Autogenous Control of rpsO |
title_short | Regulation of Ribosomal Protein Synthesis in Mycobacteria: The Autogenous Control of rpsO |
title_sort | regulation of ribosomal protein synthesis in mycobacteria: the autogenous control of rpso |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8470358/ https://www.ncbi.nlm.nih.gov/pubmed/34575857 http://dx.doi.org/10.3390/ijms22189679 |
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