The RNase J-Based RNA Degradosome Is Compartmentalized in the Gastric Pathogen <named-content content-type="genus-species">Helicobacter pylori</named-content>

ABSTRACT Posttranscriptional regulation is a major level of gene expression control in any cell. In bacteria, multiprotein machines called RNA degradosomes are central for RNA processing and degradation, and some were reported to be compartmentalized inside these organelleless cells. The minimal RNA...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Alejandro Tejada-Arranz, Eloïse Galtier, Lamya El Mortaji, Evelyne Turlin, Dmitry Ershov, Hilde De Reuse
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	Helicobacter pylori RNA degradosome RNase J post-transcriptional regulation subcellular localization superresolution microscopy Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/247ba4ac6a1f430eb201f17d5a95753e
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:247ba4ac6a1f430eb201f17d5a95753e
record_format	dspace
spelling	oai:doaj.org-article:247ba4ac6a1f430eb201f17d5a95753e2021-11-15T16:19:07ZThe RNase J-Based RNA Degradosome Is Compartmentalized in the Gastric Pathogen <named-content content-type="genus-species">Helicobacter pylori</named-content>10.1128/mBio.01173-202150-7511https://doaj.org/article/247ba4ac6a1f430eb201f17d5a95753e2020-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01173-20https://doaj.org/toc/2150-7511ABSTRACT Posttranscriptional regulation is a major level of gene expression control in any cell. In bacteria, multiprotein machines called RNA degradosomes are central for RNA processing and degradation, and some were reported to be compartmentalized inside these organelleless cells. The minimal RNA degradosome of the important gastric pathogen Helicobacter pylori is composed of the essential ribonuclease RNase J and RhpA, its sole DEAD box RNA helicase, and plays a major role in the regulation of mRNA decay and adaptation to gastric colonization. Here, the subcellular localization of the H. pylori RNA degradosome was investigated using cellular fractionation and both confocal and superresolution microscopy. We established that RNase J and RhpA are peripheral inner membrane proteins and that this association was mediated neither by ribosomes nor by RNA nor by the RNase Y membrane protein. In live H. pylori cells, we observed that fluorescent RNase J and RhpA protein fusions assemble into nonpolar foci. We identified factors that regulate the formation of these foci without affecting the degradosome membrane association. Flotillin, a bacterial membrane scaffolding protein, and free RNA promote focus formation in H. pylori. Finally, RNase J-GFP (RNase J-green fluorescent protein) molecules and foci in cells were quantified by three-dimensional (3D) single-molecule fluorescence localization microscopy. The number and size of the RNase J foci were found to be scaled with growth phase and cell volume as previously reported for eukaryotic ribonucleoprotein granules. In conclusion, we propose that membrane compartmentalization and the regulated clustering of RNase J-based degradosome hubs represent important levels of control of their activity and specificity. IMPORTANCE Helicobacter pylori is a bacterial pathogen that chronically colonizes the stomach of half of the human population worldwide. Infection by H. pylori can lead to the development of gastric pathologies such as ulcers and adenocarcinoma, which causes up to 800,000 deaths in the world each year. Persistent colonization by H. pylori relies on regulation of the expression of adaptation-related genes. One major level of such control is posttranscriptional regulation, which, in H. pylori, largely relies on a multiprotein molecular machine, an RNA degradosome, that we previously discovered. In this study, we established that the two protein partners of this machine are associated with the membrane of H. pylori. Using cutting-edge microscopy, we showed that these complexes assemble into hubs whose formation is regulated by free RNA and scaled with bacterial size and growth phase. Organelleless cellular compartmentalization of molecular machines into hubs emerges as an important regulatory level in bacteria.Alejandro Tejada-ArranzEloïse GaltierLamya El MortajiEvelyne TurlinDmitry ErshovHilde De ReuseAmerican Society for MicrobiologyarticleHelicobacter pyloriRNA degradosomeRNase Jpost-transcriptional regulationsubcellular localizationsuperresolution microscopyMicrobiologyQR1-502ENmBio, Vol 11, Iss 5 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Helicobacter pylori RNA degradosome RNase J post-transcriptional regulation subcellular localization superresolution microscopy Microbiology QR1-502
spellingShingle	Helicobacter pylori RNA degradosome RNase J post-transcriptional regulation subcellular localization superresolution microscopy Microbiology QR1-502 Alejandro Tejada-Arranz Eloïse Galtier Lamya El Mortaji Evelyne Turlin Dmitry Ershov Hilde De Reuse The RNase J-Based RNA Degradosome Is Compartmentalized in the Gastric Pathogen <named-content content-type="genus-species">Helicobacter pylori</named-content>
description	ABSTRACT Posttranscriptional regulation is a major level of gene expression control in any cell. In bacteria, multiprotein machines called RNA degradosomes are central for RNA processing and degradation, and some were reported to be compartmentalized inside these organelleless cells. The minimal RNA degradosome of the important gastric pathogen Helicobacter pylori is composed of the essential ribonuclease RNase J and RhpA, its sole DEAD box RNA helicase, and plays a major role in the regulation of mRNA decay and adaptation to gastric colonization. Here, the subcellular localization of the H. pylori RNA degradosome was investigated using cellular fractionation and both confocal and superresolution microscopy. We established that RNase J and RhpA are peripheral inner membrane proteins and that this association was mediated neither by ribosomes nor by RNA nor by the RNase Y membrane protein. In live H. pylori cells, we observed that fluorescent RNase J and RhpA protein fusions assemble into nonpolar foci. We identified factors that regulate the formation of these foci without affecting the degradosome membrane association. Flotillin, a bacterial membrane scaffolding protein, and free RNA promote focus formation in H. pylori. Finally, RNase J-GFP (RNase J-green fluorescent protein) molecules and foci in cells were quantified by three-dimensional (3D) single-molecule fluorescence localization microscopy. The number and size of the RNase J foci were found to be scaled with growth phase and cell volume as previously reported for eukaryotic ribonucleoprotein granules. In conclusion, we propose that membrane compartmentalization and the regulated clustering of RNase J-based degradosome hubs represent important levels of control of their activity and specificity. IMPORTANCE Helicobacter pylori is a bacterial pathogen that chronically colonizes the stomach of half of the human population worldwide. Infection by H. pylori can lead to the development of gastric pathologies such as ulcers and adenocarcinoma, which causes up to 800,000 deaths in the world each year. Persistent colonization by H. pylori relies on regulation of the expression of adaptation-related genes. One major level of such control is posttranscriptional regulation, which, in H. pylori, largely relies on a multiprotein molecular machine, an RNA degradosome, that we previously discovered. In this study, we established that the two protein partners of this machine are associated with the membrane of H. pylori. Using cutting-edge microscopy, we showed that these complexes assemble into hubs whose formation is regulated by free RNA and scaled with bacterial size and growth phase. Organelleless cellular compartmentalization of molecular machines into hubs emerges as an important regulatory level in bacteria.
format	article
author	Alejandro Tejada-Arranz Eloïse Galtier Lamya El Mortaji Evelyne Turlin Dmitry Ershov Hilde De Reuse
author_facet	Alejandro Tejada-Arranz Eloïse Galtier Lamya El Mortaji Evelyne Turlin Dmitry Ershov Hilde De Reuse
author_sort	Alejandro Tejada-Arranz
title	The RNase J-Based RNA Degradosome Is Compartmentalized in the Gastric Pathogen <named-content content-type="genus-species">Helicobacter pylori</named-content>
title_short	The RNase J-Based RNA Degradosome Is Compartmentalized in the Gastric Pathogen <named-content content-type="genus-species">Helicobacter pylori</named-content>
title_full	The RNase J-Based RNA Degradosome Is Compartmentalized in the Gastric Pathogen <named-content content-type="genus-species">Helicobacter pylori</named-content>
title_fullStr	The RNase J-Based RNA Degradosome Is Compartmentalized in the Gastric Pathogen <named-content content-type="genus-species">Helicobacter pylori</named-content>
title_full_unstemmed	The RNase J-Based RNA Degradosome Is Compartmentalized in the Gastric Pathogen <named-content content-type="genus-species">Helicobacter pylori</named-content>
title_sort	rnase j-based rna degradosome is compartmentalized in the gastric pathogen <named-content content-type="genus-species">helicobacter pylori</named-content>
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/247ba4ac6a1f430eb201f17d5a95753e
work_keys_str_mv	AT alejandrotejadaarranz thernasejbasedrnadegradosomeiscompartmentalizedinthegastricpathogennamedcontentcontenttypegenusspecieshelicobacterpylorinamedcontent AT eloisegaltier thernasejbasedrnadegradosomeiscompartmentalizedinthegastricpathogennamedcontentcontenttypegenusspecieshelicobacterpylorinamedcontent AT lamyaelmortaji thernasejbasedrnadegradosomeiscompartmentalizedinthegastricpathogennamedcontentcontenttypegenusspecieshelicobacterpylorinamedcontent AT evelyneturlin thernasejbasedrnadegradosomeiscompartmentalizedinthegastricpathogennamedcontentcontenttypegenusspecieshelicobacterpylorinamedcontent AT dmitryershov thernasejbasedrnadegradosomeiscompartmentalizedinthegastricpathogennamedcontentcontenttypegenusspecieshelicobacterpylorinamedcontent AT hildedereuse thernasejbasedrnadegradosomeiscompartmentalizedinthegastricpathogennamedcontentcontenttypegenusspecieshelicobacterpylorinamedcontent AT alejandrotejadaarranz rnasejbasedrnadegradosomeiscompartmentalizedinthegastricpathogennamedcontentcontenttypegenusspecieshelicobacterpylorinamedcontent AT eloisegaltier rnasejbasedrnadegradosomeiscompartmentalizedinthegastricpathogennamedcontentcontenttypegenusspecieshelicobacterpylorinamedcontent AT lamyaelmortaji rnasejbasedrnadegradosomeiscompartmentalizedinthegastricpathogennamedcontentcontenttypegenusspecieshelicobacterpylorinamedcontent AT evelyneturlin rnasejbasedrnadegradosomeiscompartmentalizedinthegastricpathogennamedcontentcontenttypegenusspecieshelicobacterpylorinamedcontent AT dmitryershov rnasejbasedrnadegradosomeiscompartmentalizedinthegastricpathogennamedcontentcontenttypegenusspecieshelicobacterpylorinamedcontent AT hildedereuse rnasejbasedrnadegradosomeiscompartmentalizedinthegastricpathogennamedcontentcontenttypegenusspecieshelicobacterpylorinamedcontent
_version_	1718426954587176960

The RNase J-Based RNA Degradosome Is Compartmentalized in the Gastric Pathogen <named-content content-type="genus-species">Helicobacter pylori</named-content>

Ejemplares similares