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Deciphering the Regulatory Circuits of RA3 Replication Module - Mechanisms of the Copy Number Control
The RA3 plasmid, the archetype of IncU incompatibility group, represents a mosaic-modular genome of 45.9 kb. The replication module encompasses repA and repB (initiator) surrounded by two long repetitive sequences DR1 and DR2 of unknown function. Here, we mapped the origin of replication oriV to the...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9455977/ https://www.ncbi.nlm.nih.gov/pubmed/36077372 http://dx.doi.org/10.3390/ijms23179964 |
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author | Markowska-Barkic, Aleksandra Lewicka, Ewa Czeredys, Magdalena Mitura, Monika Jagura-Burdzy, Grazyna |
author_facet | Markowska-Barkic, Aleksandra Lewicka, Ewa Czeredys, Magdalena Mitura, Monika Jagura-Burdzy, Grazyna |
author_sort | Markowska-Barkic, Aleksandra |
collection | PubMed |
description | The RA3 plasmid, the archetype of IncU incompatibility group, represents a mosaic-modular genome of 45.9 kb. The replication module encompasses repA and repB (initiator) surrounded by two long repetitive sequences DR1 and DR2 of unknown function. Here, we mapped the origin of replication oriV to the 3′ end of repB and showed that oriV was activated by the transcription coming from orf02revp in the adjacent stability module. Using various in vivo and in vitro methods we demonstrated that the repB expression proceeded either from repBp located in the intergenic repA-repB region or from the upstream strong repAp that was autoregulated by RepA. Additionally, the repBp activity was modulated by the transcription from the overlapping, divergently oriented repXp. Both repXmRNA (antisense for repAmRNA) and its small polypeptide product, RepX, were strong incompatibility determinants. Hence, we showed that the sophisticated RA3 copy number control combined the multivalent regulation of repB expression, RepB titration by DR1, and transcriptional activation of oriV, dependent on the RA3 global regulatory network. Similarly organized replicons have been found in diverse bacterial species confirming the significance of these mechanisms in establishing the IncU plasmids in a broad spectrum of hosts. |
format | Online Article Text |
id | pubmed-9455977 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-94559772022-09-09 Deciphering the Regulatory Circuits of RA3 Replication Module - Mechanisms of the Copy Number Control Markowska-Barkic, Aleksandra Lewicka, Ewa Czeredys, Magdalena Mitura, Monika Jagura-Burdzy, Grazyna Int J Mol Sci Article The RA3 plasmid, the archetype of IncU incompatibility group, represents a mosaic-modular genome of 45.9 kb. The replication module encompasses repA and repB (initiator) surrounded by two long repetitive sequences DR1 and DR2 of unknown function. Here, we mapped the origin of replication oriV to the 3′ end of repB and showed that oriV was activated by the transcription coming from orf02revp in the adjacent stability module. Using various in vivo and in vitro methods we demonstrated that the repB expression proceeded either from repBp located in the intergenic repA-repB region or from the upstream strong repAp that was autoregulated by RepA. Additionally, the repBp activity was modulated by the transcription from the overlapping, divergently oriented repXp. Both repXmRNA (antisense for repAmRNA) and its small polypeptide product, RepX, were strong incompatibility determinants. Hence, we showed that the sophisticated RA3 copy number control combined the multivalent regulation of repB expression, RepB titration by DR1, and transcriptional activation of oriV, dependent on the RA3 global regulatory network. Similarly organized replicons have been found in diverse bacterial species confirming the significance of these mechanisms in establishing the IncU plasmids in a broad spectrum of hosts. MDPI 2022-09-01 /pmc/articles/PMC9455977/ /pubmed/36077372 http://dx.doi.org/10.3390/ijms23179964 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Markowska-Barkic, Aleksandra Lewicka, Ewa Czeredys, Magdalena Mitura, Monika Jagura-Burdzy, Grazyna Deciphering the Regulatory Circuits of RA3 Replication Module - Mechanisms of the Copy Number Control |
title | Deciphering the Regulatory Circuits of RA3 Replication Module - Mechanisms of the Copy Number Control |
title_full | Deciphering the Regulatory Circuits of RA3 Replication Module - Mechanisms of the Copy Number Control |
title_fullStr | Deciphering the Regulatory Circuits of RA3 Replication Module - Mechanisms of the Copy Number Control |
title_full_unstemmed | Deciphering the Regulatory Circuits of RA3 Replication Module - Mechanisms of the Copy Number Control |
title_short | Deciphering the Regulatory Circuits of RA3 Replication Module - Mechanisms of the Copy Number Control |
title_sort | deciphering the regulatory circuits of ra3 replication module - mechanisms of the copy number control |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9455977/ https://www.ncbi.nlm.nih.gov/pubmed/36077372 http://dx.doi.org/10.3390/ijms23179964 |
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