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An RNA-dependent mechanism for transient expression of bacterial translocation filaments
The prokaryotic RNA chaperone Hfq mediates sRNA–mRNA interactions and plays a significant role in post-transcriptional regulation of the type III secretion (T3S) system produced by a range of Escherichia coli pathotypes. UV-crosslinking was used to map Hfq-binding under conditions that promote T3S a...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5909449/ https://www.ncbi.nlm.nih.gov/pubmed/29432565 http://dx.doi.org/10.1093/nar/gky096 |
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author | Wang, Dai McAteer, Sean P Wawszczyk, Agata B Russell, Clark D Tahoun, Amin Elmi, Alex Cockroft, Scott L Tollervey, David Granneman, Sander Tree, Jai J Gally, David L |
author_facet | Wang, Dai McAteer, Sean P Wawszczyk, Agata B Russell, Clark D Tahoun, Amin Elmi, Alex Cockroft, Scott L Tollervey, David Granneman, Sander Tree, Jai J Gally, David L |
author_sort | Wang, Dai |
collection | PubMed |
description | The prokaryotic RNA chaperone Hfq mediates sRNA–mRNA interactions and plays a significant role in post-transcriptional regulation of the type III secretion (T3S) system produced by a range of Escherichia coli pathotypes. UV-crosslinking was used to map Hfq-binding under conditions that promote T3S and multiple interactions were identified within polycistronic transcripts produced from the locus of enterocyte effacement (LEE) that encodes the T3S system. The majority of Hfq binding was within the LEE5 and LEE4 operons, the latter encoding the translocon apparatus (SepL-EspADB) that is positively regulated by the RNA binding protein, CsrA. Using the identified Hfq-binding sites and a series of sRNA deletions, the sRNA Spot42 was shown to directly repress translation of LEE4 at the sepL 5′ UTR. In silico and in vivo analyses of the sepL mRNA secondary structure combined with expression studies of truncates indicated that the unbound sepL mRNA is translationally inactive. Based on expression studies with site-directed mutants, an OFF-ON-OFF toggle model is proposed that results in transient translation of SepL and EspA filament assembly. Under this model, the nascent mRNA is translationally off, before being activated by CsrA, and then repressed by Hfq and Spot42. |
format | Online Article Text |
id | pubmed-5909449 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-59094492018-04-24 An RNA-dependent mechanism for transient expression of bacterial translocation filaments Wang, Dai McAteer, Sean P Wawszczyk, Agata B Russell, Clark D Tahoun, Amin Elmi, Alex Cockroft, Scott L Tollervey, David Granneman, Sander Tree, Jai J Gally, David L Nucleic Acids Res Gene regulation, Chromatin and Epigenetics The prokaryotic RNA chaperone Hfq mediates sRNA–mRNA interactions and plays a significant role in post-transcriptional regulation of the type III secretion (T3S) system produced by a range of Escherichia coli pathotypes. UV-crosslinking was used to map Hfq-binding under conditions that promote T3S and multiple interactions were identified within polycistronic transcripts produced from the locus of enterocyte effacement (LEE) that encodes the T3S system. The majority of Hfq binding was within the LEE5 and LEE4 operons, the latter encoding the translocon apparatus (SepL-EspADB) that is positively regulated by the RNA binding protein, CsrA. Using the identified Hfq-binding sites and a series of sRNA deletions, the sRNA Spot42 was shown to directly repress translation of LEE4 at the sepL 5′ UTR. In silico and in vivo analyses of the sepL mRNA secondary structure combined with expression studies of truncates indicated that the unbound sepL mRNA is translationally inactive. Based on expression studies with site-directed mutants, an OFF-ON-OFF toggle model is proposed that results in transient translation of SepL and EspA filament assembly. Under this model, the nascent mRNA is translationally off, before being activated by CsrA, and then repressed by Hfq and Spot42. Oxford University Press 2018-04-20 2018-02-08 /pmc/articles/PMC5909449/ /pubmed/29432565 http://dx.doi.org/10.1093/nar/gky096 Text en © The Author(s) 2018. 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 | Gene regulation, Chromatin and Epigenetics Wang, Dai McAteer, Sean P Wawszczyk, Agata B Russell, Clark D Tahoun, Amin Elmi, Alex Cockroft, Scott L Tollervey, David Granneman, Sander Tree, Jai J Gally, David L An RNA-dependent mechanism for transient expression of bacterial translocation filaments |
title | An RNA-dependent mechanism for transient expression of bacterial translocation filaments |
title_full | An RNA-dependent mechanism for transient expression of bacterial translocation filaments |
title_fullStr | An RNA-dependent mechanism for transient expression of bacterial translocation filaments |
title_full_unstemmed | An RNA-dependent mechanism for transient expression of bacterial translocation filaments |
title_short | An RNA-dependent mechanism for transient expression of bacterial translocation filaments |
title_sort | rna-dependent mechanism for transient expression of bacterial translocation filaments |
topic | Gene regulation, Chromatin and Epigenetics |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5909449/ https://www.ncbi.nlm.nih.gov/pubmed/29432565 http://dx.doi.org/10.1093/nar/gky096 |
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