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RNase J1 and J2 Are Host-Encoded Factors for Plasmid Replication

Plasmids need to ensure their transmission to both daughter-cells when their host divides, but should at the same time avoid overtaxing their hosts by directing excessive host-resources toward production of plasmid factors. Naturally occurring plasmids have therefore evolved regulatory mechanisms to...

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Autores principales: Guimarães, Vanessa Andrade, Le Scornet, Alexandre, Khemici, Vanessa, Hausmann, Stéphane, Armitano, Joshua, Prados, Julien, Jousselin, Ambre, Manzano, Caroline, Linder, Patrick, Redder, Peter
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8129170/
https://www.ncbi.nlm.nih.gov/pubmed/34017314
http://dx.doi.org/10.3389/fmicb.2021.586886
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author Guimarães, Vanessa Andrade
Le Scornet, Alexandre
Khemici, Vanessa
Hausmann, Stéphane
Armitano, Joshua
Prados, Julien
Jousselin, Ambre
Manzano, Caroline
Linder, Patrick
Redder, Peter
author_facet Guimarães, Vanessa Andrade
Le Scornet, Alexandre
Khemici, Vanessa
Hausmann, Stéphane
Armitano, Joshua
Prados, Julien
Jousselin, Ambre
Manzano, Caroline
Linder, Patrick
Redder, Peter
author_sort Guimarães, Vanessa Andrade
collection PubMed
description Plasmids need to ensure their transmission to both daughter-cells when their host divides, but should at the same time avoid overtaxing their hosts by directing excessive host-resources toward production of plasmid factors. Naturally occurring plasmids have therefore evolved regulatory mechanisms to restrict their copy-number in response to the volume of the cytoplasm. In many plasmid families, copy-number control is mediated by a small plasmid-specified RNA, which is continuously produced and rapidly degraded, to ensure that its concentration is proportional to the current plasmid copy-number. We show here that pSA564 from the RepA_N-family is regulated by a small antisense RNA (RNA1), which, when over-expressed in trans, blocks plasmid replication and cures the bacterial host. The 5′ untranslated region (5′UTR) of the plasmid replication initiation gene (repA) potentially forms two mutually exclusive secondary structures, ON and OFF, where the latter both sequesters the repA ribosome binding site and acts as a rho-independent transcriptional terminator. Duplex formation between RNA1 and the 5′UTR shifts the equilibrium to favor the putative OFF-structure, enabling a single small RNA to down-regulate repA expression at both transcriptional and translational levels. We further examine which sequence elements on the antisense RNA and on its 5′UTR target are needed for this regulation. Finally, we identify the host-encoded exoribonucleases RNase J1 and J2 as the enzymes responsible for rapidly degrading the replication-inhibiting section of RNA1. This region accumulates and blocks RepA expression in the absence of either RNase J1 or J2, which are therefore essential host factors for pSA564 replication in Staphylococcus aureus.
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spelling pubmed-81291702021-05-19 RNase J1 and J2 Are Host-Encoded Factors for Plasmid Replication Guimarães, Vanessa Andrade Le Scornet, Alexandre Khemici, Vanessa Hausmann, Stéphane Armitano, Joshua Prados, Julien Jousselin, Ambre Manzano, Caroline Linder, Patrick Redder, Peter Front Microbiol Microbiology Plasmids need to ensure their transmission to both daughter-cells when their host divides, but should at the same time avoid overtaxing their hosts by directing excessive host-resources toward production of plasmid factors. Naturally occurring plasmids have therefore evolved regulatory mechanisms to restrict their copy-number in response to the volume of the cytoplasm. In many plasmid families, copy-number control is mediated by a small plasmid-specified RNA, which is continuously produced and rapidly degraded, to ensure that its concentration is proportional to the current plasmid copy-number. We show here that pSA564 from the RepA_N-family is regulated by a small antisense RNA (RNA1), which, when over-expressed in trans, blocks plasmid replication and cures the bacterial host. The 5′ untranslated region (5′UTR) of the plasmid replication initiation gene (repA) potentially forms two mutually exclusive secondary structures, ON and OFF, where the latter both sequesters the repA ribosome binding site and acts as a rho-independent transcriptional terminator. Duplex formation between RNA1 and the 5′UTR shifts the equilibrium to favor the putative OFF-structure, enabling a single small RNA to down-regulate repA expression at both transcriptional and translational levels. We further examine which sequence elements on the antisense RNA and on its 5′UTR target are needed for this regulation. Finally, we identify the host-encoded exoribonucleases RNase J1 and J2 as the enzymes responsible for rapidly degrading the replication-inhibiting section of RNA1. This region accumulates and blocks RepA expression in the absence of either RNase J1 or J2, which are therefore essential host factors for pSA564 replication in Staphylococcus aureus. Frontiers Media S.A. 2021-05-04 /pmc/articles/PMC8129170/ /pubmed/34017314 http://dx.doi.org/10.3389/fmicb.2021.586886 Text en Copyright © 2021 Guimarães, Le Scornet, Khemici, Hausmann, Armitano, Prados, Jousselin, Manzano, Linder and Redder. https://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) and the copyright owner(s) 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
Guimarães, Vanessa Andrade
Le Scornet, Alexandre
Khemici, Vanessa
Hausmann, Stéphane
Armitano, Joshua
Prados, Julien
Jousselin, Ambre
Manzano, Caroline
Linder, Patrick
Redder, Peter
RNase J1 and J2 Are Host-Encoded Factors for Plasmid Replication
title RNase J1 and J2 Are Host-Encoded Factors for Plasmid Replication
title_full RNase J1 and J2 Are Host-Encoded Factors for Plasmid Replication
title_fullStr RNase J1 and J2 Are Host-Encoded Factors for Plasmid Replication
title_full_unstemmed RNase J1 and J2 Are Host-Encoded Factors for Plasmid Replication
title_short RNase J1 and J2 Are Host-Encoded Factors for Plasmid Replication
title_sort rnase j1 and j2 are host-encoded factors for plasmid replication
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8129170/
https://www.ncbi.nlm.nih.gov/pubmed/34017314
http://dx.doi.org/10.3389/fmicb.2021.586886
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