Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Markowska-Barkic, Aleksandra, Lewicka, Ewa, Czeredys, Magdalena, Mitura, Monika, Jagura-Burdzy, Grazyna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
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
_version_ 1784785697538834432
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
work_keys_str_mv AT markowskabarkicaleksandra decipheringtheregulatorycircuitsofra3replicationmodulemechanismsofthecopynumbercontrol
AT lewickaewa decipheringtheregulatorycircuitsofra3replicationmodulemechanismsofthecopynumbercontrol
AT czeredysmagdalena decipheringtheregulatorycircuitsofra3replicationmodulemechanismsofthecopynumbercontrol
AT mituramonika decipheringtheregulatorycircuitsofra3replicationmodulemechanismsofthecopynumbercontrol
AT jaguraburdzygrazyna decipheringtheregulatorycircuitsofra3replicationmodulemechanismsofthecopynumbercontrol