Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Aseev, Leonid V., Koledinskaya, Ludmila S., Bychenko, Oksana S., Boni, Irina V.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
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
_version_ 1784574178642362368
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
work_keys_str_mv AT aseevleonidv regulationofribosomalproteinsynthesisinmycobacteriatheautogenouscontrolofrpso
AT koledinskayaludmilas regulationofribosomalproteinsynthesisinmycobacteriatheautogenouscontrolofrpso
AT bychenkooksanas regulationofribosomalproteinsynthesisinmycobacteriatheautogenouscontrolofrpso
AT boniirinav regulationofribosomalproteinsynthesisinmycobacteriatheautogenouscontrolofrpso