Plasmid R6K replication control

Rakowski, Sheryl A.; Filutowicz, Marcin

doi:10.1016/j.plasmid.2013.02.003

Cited by 52 publications

(37 citation statements)

References 115 publications

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“…5). Both SUSP1 and SUSP2 efficiently liberated two broad-host-range plasmids, pOAR31 and pπγ, whose environmental counterparts are prominent vectors of virulence factors and antibiotic resistance genes (5, 34, 35). Ultimately, the biological relevance of superspreading phages is likely to be principally regulated by the host ranges of their liberated plasmids, by the relative abundance of naturally competent bacteria compatible with those plasmids, and by factors that influence the persistence of DNA in natural environments.…”

Section: Discussionmentioning

confidence: 99%

Novel “Superspreader” Bacteriophages Promote Horizontal Gene Transfer by Transformation

Keen

Bliskovsky

Malagón

et al. 2017

mBio

117

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Bacteriophages infect an estimated 1023 to 1025 bacterial cells each second, many of which carry physiologically relevant plasmids (e.g., those encoding antibiotic resistance). However, even though phage-plasmid interactions occur on a massive scale and have potentially significant evolutionary, ecological, and biomedical implications, plasmid fate upon phage infection and lysis has not been investigated to date. Here we show that a subset of the natural lytic phage population, which we dub “superspreaders,” releases substantial amounts of intact, transformable plasmid DNA upon lysis, thereby promoting horizontal gene transfer by transformation. Two novel Escherichia coli phage superspreaders, SUSP1 and SUSP2, liberated four evolutionarily distinct plasmids with equal efficiency, including two close relatives of prominent antibiotic resistance vectors in natural environments. SUSP2 also mediated the extensive lateral transfer of antibiotic resistance in unbiased communities of soil bacteria from Maryland and Wyoming. Furthermore, the addition of SUSP2 to cocultures of kanamycin-resistant E. coli and kanamycin-sensitive Bacillus sp. bacteria resulted in roughly 1,000-fold more kanamycin-resistant Bacillus sp. bacteria than arose in phage-free controls. Unlike many other lytic phages, neither SUSP1 nor SUSP2 encodes homologs to known hydrolytic endonucleases, suggesting a simple potential mechanism underlying the superspreading phenotype. Consistent with this model, the deletion of endonuclease IV and the nucleoid-disrupting protein ndd from coliphage T4, a phage known to extensively degrade chromosomal DNA, significantly increased its ability to promote plasmid transformation. Taken together, our results suggest that phage superspreaders may play key roles in microbial evolution and ecology but should be avoided in phage therapy and other medical applications.

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Section: Discussionmentioning

confidence: 99%

Novel “Superspreader” Bacteriophages Promote Horizontal Gene Transfer by Transformation

Keen

Bliskovsky

Malagón

et al. 2017

mBio

117

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“…The replication protein is encoded by pir (protein for initiation of replication), and X plasmids have three ori regions: ␥ (part of the minimal replicon), ␣, and ␤ (287). Replication is regulated by handcuffing mediated by dimers coupling two ori␥ regions (148,287). The conjugation region consists of genes for pilus synthesis and assembly (originally named pilX1 to -11 but renamed tivB1 to -11) plus taxC/rlxX (relaxase gene), taxB/cplX (coupling protein gene), and taxA/dtrX1 (auxiliary relaxosome protein gene) (286).…”

Section: Resistance Plasmids In the Enterobacteriaceaementioning

confidence: 99%

Mobile Genetic Elements Associated with Antimicrobial Resistance

et al. 2018

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SUMMARYStrains of bacteria resistant to antibiotics, particularly those that are multiresistant, are an increasing major health care problem around the world. It is now abundantly clear that both Gram-negative and Gram-positive bacteria are able to meet the evolutionary challenge of combating antimicrobial chemotherapy, often by acquiring preexisting resistance determinants from the bacterial gene pool. This is achieved through the concerted activities of mobile genetic elements able to move within or between DNA molecules, which include insertion sequences, transposons, and gene cassettes/integrons, and those that are able to transfer between bacterial cells, such as plasmids and integrative conjugative elements. Together these elements play a central role in facilitating horizontal genetic exchange and therefore promote the acquisition and spread of resistance genes. This review aims to outline the characteristics of the major types of mobile genetic elements involved in acquisition and spread of antibiotic resistance in both Gram-negative and Gram-positive bacteria, focusing on the so-called ESKAPEE group of organisms (, ,, ,, spp., and), which have become the most problematic hospital pathogens.

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“…To allow tuning of the duration of differentiated cell proliferation, we take advantage of the reliance of R6K plasmid replication on the  protein encoded by the pir gene. 20 By varying the expression of the pir gene using an inducible promoter, and using the recombination event to inactivate its expression, the  protein abundance, R6K plasmid copy number, and therefor the number of cell divisions differentiated cells undergo before losing the R6K plasmid and its associated antibiotic resistance gene may be tuned (Fig. 3A).…”

Section: Resultsmentioning

confidence: 99%

Integrase-mediated differentiation circuits improve evolutionary stability of burdensome and toxic functions in E. coli

Williams

Murray

2019

Preprint

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Application of synthetic biology is limited by the capacity of cells to faithfully execute burdensome engineered functions in the face of Darwinian evolution. Division of labor, both metabolic and reproductive, are underutilized in confronting this barrier. To address this, we developed a serine-integrase based differentiation circuit that allows control of the population composition through tuning of the differentiation rate and number of cell divisions differentiated cells can undergo. We applied this system to T7 RNAP-driven expression of a fluorescent protein, and demonstrate both increased duration of circuit function and total production for high burden expression. While T7 expression systems are typically used for high-level short-term expression, this system enables longer duration production, and could be readily applied to burdensome or toxic products not readily produced in bacteria.

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Plasmid R6K replication control

Cited by 52 publications

References 115 publications

Novel “Superspreader” Bacteriophages Promote Horizontal Gene Transfer by Transformation

Novel “Superspreader” Bacteriophages Promote Horizontal Gene Transfer by Transformation

Mobile Genetic Elements Associated with Antimicrobial Resistance

Integrase-mediated differentiation circuits improve evolutionary stability of burdensome and toxic functions in E. coli

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