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Small protein B upregulates sensor kinase bvgS expression in Aeromonas veronii
Earlier studies reveal that Small protein B (SmpB), a class of well-conserved tmRNA-binding proteins, is essential for the trans-translation process, which functions as a system for translation surveillance and ribosome rescue. Here, we report a previously unrecognized mechanism by which SmpB alone...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4468919/ https://www.ncbi.nlm.nih.gov/pubmed/26136727 http://dx.doi.org/10.3389/fmicb.2015.00579 |
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author | Liu, Zhu Liu, Peng Liu, Shuanshuan Song, Haichao Tang, Hongqian Hu, Xinwen |
author_facet | Liu, Zhu Liu, Peng Liu, Shuanshuan Song, Haichao Tang, Hongqian Hu, Xinwen |
author_sort | Liu, Zhu |
collection | PubMed |
description | Earlier studies reveal that Small protein B (SmpB), a class of well-conserved tmRNA-binding proteins, is essential for the trans-translation process, which functions as a system for translation surveillance and ribosome rescue. Here, we report a previously unrecognized mechanism by which SmpB alone positively regulates the expression of a sensor kinase, BvgS, in Aeromonas veronii. A reporter plasmid was constructed in which the promoter of bvgS was used to control the expression of the enhanced green fluorescent protein (eGFP) gene. When the reporter plasmid was co-transformed with a SmpB expression construct into E. coli, the relative fluorescence intensity increased about threefold. Transformation with a truncated form of smpB gene showed that the C-terminus had little effect, while N-terminus unexpectedly increased eGFP production. Next, a series of SmpB mutants were generated by site-directed mutagenesis. When the mutants SmpB (G11S) or SmpB (E32AG) was used in the experiment, eGFP expression dropped significantly compared with that of wild type SmpB. Further, purified SmpB was shown to bind the promoter regions of bvgS in the agarose gel retardation assay. Quantitative RT-PCR analysis showed that eGFP transcript levels increased approximately 25-fold in the presence of SmpB. Likewise, smpB knockout decreased bvgS transcripts significantly in A. veronii, and also displayed a reduced capability in salt tolerance. Collectively, the data presented here will facilitate a deeper understanding of SmpB-mediated regulatory circuits as a transcriptional factor in A. veronii. |
format | Online Article Text |
id | pubmed-4468919 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-44689192015-07-01 Small protein B upregulates sensor kinase bvgS expression in Aeromonas veronii Liu, Zhu Liu, Peng Liu, Shuanshuan Song, Haichao Tang, Hongqian Hu, Xinwen Front Microbiol Microbiology Earlier studies reveal that Small protein B (SmpB), a class of well-conserved tmRNA-binding proteins, is essential for the trans-translation process, which functions as a system for translation surveillance and ribosome rescue. Here, we report a previously unrecognized mechanism by which SmpB alone positively regulates the expression of a sensor kinase, BvgS, in Aeromonas veronii. A reporter plasmid was constructed in which the promoter of bvgS was used to control the expression of the enhanced green fluorescent protein (eGFP) gene. When the reporter plasmid was co-transformed with a SmpB expression construct into E. coli, the relative fluorescence intensity increased about threefold. Transformation with a truncated form of smpB gene showed that the C-terminus had little effect, while N-terminus unexpectedly increased eGFP production. Next, a series of SmpB mutants were generated by site-directed mutagenesis. When the mutants SmpB (G11S) or SmpB (E32AG) was used in the experiment, eGFP expression dropped significantly compared with that of wild type SmpB. Further, purified SmpB was shown to bind the promoter regions of bvgS in the agarose gel retardation assay. Quantitative RT-PCR analysis showed that eGFP transcript levels increased approximately 25-fold in the presence of SmpB. Likewise, smpB knockout decreased bvgS transcripts significantly in A. veronii, and also displayed a reduced capability in salt tolerance. Collectively, the data presented here will facilitate a deeper understanding of SmpB-mediated regulatory circuits as a transcriptional factor in A. veronii. Frontiers Media S.A. 2015-06-16 /pmc/articles/PMC4468919/ /pubmed/26136727 http://dx.doi.org/10.3389/fmicb.2015.00579 Text en Copyright © 2015 Liu, Liu, Liu, Song, Tang and Hu. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Microbiology Liu, Zhu Liu, Peng Liu, Shuanshuan Song, Haichao Tang, Hongqian Hu, Xinwen Small protein B upregulates sensor kinase bvgS expression in Aeromonas veronii |
title | Small protein B upregulates sensor kinase bvgS expression in Aeromonas veronii |
title_full | Small protein B upregulates sensor kinase bvgS expression in Aeromonas veronii |
title_fullStr | Small protein B upregulates sensor kinase bvgS expression in Aeromonas veronii |
title_full_unstemmed | Small protein B upregulates sensor kinase bvgS expression in Aeromonas veronii |
title_short | Small protein B upregulates sensor kinase bvgS expression in Aeromonas veronii |
title_sort | small protein b upregulates sensor kinase bvgs expression in aeromonas veronii |
topic | Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4468919/ https://www.ncbi.nlm.nih.gov/pubmed/26136727 http://dx.doi.org/10.3389/fmicb.2015.00579 |
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