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Nucleoid and cytoplasmic localization of small RNAs in Escherichia coli
Bacterial small RNAs (sRNAs) regulate protein production by binding to mRNAs and altering their translation and degradation. sRNAs are smaller than most mRNAs but larger than many proteins. Therefore it is uncertain whether sRNAs can enter the nucleoid to target nascent mRNAs. Here, we investigate t...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5389542/ https://www.ncbi.nlm.nih.gov/pubmed/28119418 http://dx.doi.org/10.1093/nar/gkx023 |
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author | Sheng, Huanjie Stauffer, Weston T. Hussein, Razika Lin, Chris Lim, Han N. |
author_facet | Sheng, Huanjie Stauffer, Weston T. Hussein, Razika Lin, Chris Lim, Han N. |
author_sort | Sheng, Huanjie |
collection | PubMed |
description | Bacterial small RNAs (sRNAs) regulate protein production by binding to mRNAs and altering their translation and degradation. sRNAs are smaller than most mRNAs but larger than many proteins. Therefore it is uncertain whether sRNAs can enter the nucleoid to target nascent mRNAs. Here, we investigate the intracellular localization of sRNAs transcribed from plasmids in Escherichia coli using RNA fluorescent in-situ hybridization. We found that sRNAs (GlmZ, OxyS, RyhB and SgrS) have equal preference for the nucleoid and cytoplasm, and no preferential localization at the cell membrane. We show using the gfp mRNA (encoding green fluorescent protein) that non-sRNAs can be engineered to have different proportions of nucleoid and cytoplasmic localization by altering their length and/or translation. The same localization as sRNAs was achieved by decreasing gfp mRNA length and translation, which suggests that sRNAs and other RNAs may enter the densely packed DNA of the nucleoid if they are sufficiently small. We also found that the Hfq protein, which binds sRNAs, minimally affects sRNA localization. Important implications of our findings for engineering synthetic circuits are: (i) sRNAs can potentially bind nascent mRNAs in the nucleoid, and (ii) localization patterns and distribution volumes of sRNAs can differ from some larger RNAs. |
format | Online Article Text |
id | pubmed-5389542 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-53895422017-04-24 Nucleoid and cytoplasmic localization of small RNAs in Escherichia coli Sheng, Huanjie Stauffer, Weston T. Hussein, Razika Lin, Chris Lim, Han N. Nucleic Acids Res Synthetic Biology and Bioengineering Bacterial small RNAs (sRNAs) regulate protein production by binding to mRNAs and altering their translation and degradation. sRNAs are smaller than most mRNAs but larger than many proteins. Therefore it is uncertain whether sRNAs can enter the nucleoid to target nascent mRNAs. Here, we investigate the intracellular localization of sRNAs transcribed from plasmids in Escherichia coli using RNA fluorescent in-situ hybridization. We found that sRNAs (GlmZ, OxyS, RyhB and SgrS) have equal preference for the nucleoid and cytoplasm, and no preferential localization at the cell membrane. We show using the gfp mRNA (encoding green fluorescent protein) that non-sRNAs can be engineered to have different proportions of nucleoid and cytoplasmic localization by altering their length and/or translation. The same localization as sRNAs was achieved by decreasing gfp mRNA length and translation, which suggests that sRNAs and other RNAs may enter the densely packed DNA of the nucleoid if they are sufficiently small. We also found that the Hfq protein, which binds sRNAs, minimally affects sRNA localization. Important implications of our findings for engineering synthetic circuits are: (i) sRNAs can potentially bind nascent mRNAs in the nucleoid, and (ii) localization patterns and distribution volumes of sRNAs can differ from some larger RNAs. Oxford University Press 2017-03-17 2017-01-24 /pmc/articles/PMC5389542/ /pubmed/28119418 http://dx.doi.org/10.1093/nar/gkx023 Text en © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Synthetic Biology and Bioengineering Sheng, Huanjie Stauffer, Weston T. Hussein, Razika Lin, Chris Lim, Han N. Nucleoid and cytoplasmic localization of small RNAs in Escherichia coli |
title | Nucleoid and cytoplasmic localization of small RNAs in Escherichia coli |
title_full | Nucleoid and cytoplasmic localization of small RNAs in Escherichia coli |
title_fullStr | Nucleoid and cytoplasmic localization of small RNAs in Escherichia coli |
title_full_unstemmed | Nucleoid and cytoplasmic localization of small RNAs in Escherichia coli |
title_short | Nucleoid and cytoplasmic localization of small RNAs in Escherichia coli |
title_sort | nucleoid and cytoplasmic localization of small rnas in escherichia coli |
topic | Synthetic Biology and Bioengineering |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5389542/ https://www.ncbi.nlm.nih.gov/pubmed/28119418 http://dx.doi.org/10.1093/nar/gkx023 |
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