The polyadenylase PAPI is required for virulence plasmid maintenance in pathogenic bacteria.

Publisher: Public Library of Science Country of Publication: United States NLM ID: 101238921 Publication Model: eCollection Cited Medium: Internet ISSN: 1553-7374 (Electronic) Linking ISSN: 15537366 NLM ISO Abbreviation: PLoS Pathog Subsets: MEDLINE

Original Publication: San Francisco, CA : Public Library of Science, c2005-

Plasmids*/genetics ; Plasmids*/metabolism ; Yersinia pestis*/genetics ; Yersinia pestis*/pathogenicity ; Yersinia pseudotuberculosis*/genetics ; Yersinia pseudotuberculosis*/pathogenicity ; Virulence Factors*/genetics ; Virulence Factors*/metabolism ; Bacterial Proteins*/genetics ; Bacterial Proteins*/metabolism; Animals ; Mice ; Virulence/genetics ; Type III Secretion Systems/genetics ; Type III Secretion Systems/metabolism ; Humans ; Gene Expression Regulation, Bacterial

Competing Interests: The authors have declared that no competing interests exist.
Many species of pathogenic bacteria harbor critical plasmid-encoded virulence factors, and yet the regulation of plasmid replication is often poorly understood despite playing a key role in plasmid-encoded gene expression. Human pathogenic Yersinia, including the plague agent Yersinia pestis and its close relative Y. pseudotuberculosis, require the type III secretion system (T3SS) virulence factor to subvert host defense mechanisms and colonize host tissues. The Yersinia T3SS is encoded on the IncFII plasmid for Yersinia virulence (pYV). Several layers of gene regulation enable a large increase in expression of Yersinia T3SS genes at mammalian body temperature. Surprisingly, T3SS expression is also controlled at the level of gene dosage. The number of pYV molecules relative to the number of chromosomes per cell, referred to as plasmid copy number, increases with temperature. The ability to increase and maintain elevated pYV plasmid copy number, and therefore T3SS gene dosage, at 37˚C is important for Yersinia virulence. In addition, pYV is highly stable in Yersinia at all temperatures, despite being dispensable for growth outside the host. Yet how Yersinia reinforces elevated plasmid replication and plasmid stability remains unclear. In this study, we show that the chromosomal gene pcnB encoding the polyadenylase PAP I is required for regulation of pYV plasmid copy number (PCN), maintenance of pYV in the bacterial population outside the host, robust T3SS activity, and Yersinia virulence in a mouse infection model. Likewise, pcnB/PAP I is required for robust expression of the Shigella flexneri T3SS that, similar to Yersinia, is encoded on a virulence plasmid whose replication is regulated by sRNA. Furthermore, Yersinia and Shigella pcnB/PAP I is required for maintaining model antimicrobial resistance (AMR) plasmids whose replication is regulated by sRNA, thereby increasing antibiotic resistance by ten-fold. These data suggest that pcnB/PAP I contributes to the spread and stabilization of sRNA-regulated virulence and AMR plasmids in bacterial pathogens, and is essential in maintaining the gene dosage required to mediate plasmid-encoded traits. Importantly pcnB/PAP I has been bioinformatically identified in many species of bacteria despite being studied in only a few species to date. Our work highlights the potential importance of pcnB/PAP I in antibiotic resistance, and shows for the first time that pcnB/PAP I promotes virulence plasmid stability in natural pathogenic hosts with a direct impact on bacterial virulence.
(Copyright: © 2025 Schubert et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Update of: bioRxiv. 2024 Nov 08:2024.10.11.617751. doi: 10.1101/2024.10.11.617751.. (PMID: 39416138)

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K99 AI139281 United States AI NIAID NIH HHS; R01 AI141511 United States AI NIAID NIH HHS; T32 GM007276 United States GM NIGMS NIH HHS; T32 GM008646 United States GM NIGMS NIH HHS; R01 AI128360 United States AI NIAID NIH HHS

0 (Type III Secretion Systems)
0 (Virulence Factors)
0 (Bacterial Proteins)

Date Created: 20250527 Date Completed: 20250605 Latest Revision: 20250608

20250608

PMC12140428

10.1371/journal.ppat.1012655

40424556