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Article

Deciphering the Regulatory Circuits of RA3 Replication Module - Mechanisms of the Copy Number Control

Laboratory of DNA Segregation and Life Cycle of Proteobacteria, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5a, 02-106 Warsaw, Poland
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Academic Editor: Alessandro Fatica
Int. J. Mol. Sci. 2022, 23(17), 9964; https://doi.org/10.3390/ijms23179964
Received: 6 August 2022 / Revised: 26 August 2022 / Accepted: 26 August 2022 / Published: 1 September 2022
(This article belongs to the Section Molecular Microbiology)
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. View Full-Text
Keywords: plasmid replication; RA3; IncU; RepA repressor; antisense RNA; transcriptional activation plasmid replication; RA3; IncU; RepA repressor; antisense RNA; transcriptional activation
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MDPI and ACS Style

Markowska-Barkic, A.; Lewicka, E.; Czeredys, M.; Mitura, M.; Jagura-Burdzy, G. Deciphering the Regulatory Circuits of RA3 Replication Module - Mechanisms of the Copy Number Control. Int. J. Mol. Sci. 2022, 23, 9964. https://doi.org/10.3390/ijms23179964

AMA Style

Markowska-Barkic A, Lewicka E, Czeredys M, Mitura M, Jagura-Burdzy G. Deciphering the Regulatory Circuits of RA3 Replication Module - Mechanisms of the Copy Number Control. International Journal of Molecular Sciences. 2022; 23(17):9964. https://doi.org/10.3390/ijms23179964

Chicago/Turabian Style

Markowska-Barkic, Aleksandra, Ewa Lewicka, Magdalena Czeredys, Monika Mitura, and Grazyna Jagura-Burdzy. 2022. "Deciphering the Regulatory Circuits of RA3 Replication Module - Mechanisms of the Copy Number Control" International Journal of Molecular Sciences 23, no. 17: 9964. https://doi.org/10.3390/ijms23179964

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