Comparative analysis of antirestriction activity of the ArdA and ArdB proteins encoded by genes of the R64 transmissible plasmid (IncI1)

Balabanov, V. P.; Pustovoit, K. S.; Zavilgelsky, G. B.

doi:10.1134/s0026893312010025

Cited by 6 publications

(3 citation statements)

References 15 publications

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“…This strategy enabled the identification of two genes ( ardB and 910) that played donor-specific roles, affecting conjugation from TOP10 as donor but not MG1655. ArdB has anti-restriction activity, which protects incoming plasmid DNA from restriction by the chromosomally-encoded EcoKI type I restriction-modification system [ 45 ]. EcoKI methylates recognition sites on host DNA, but cleaves unmethylated DNA from mobile genetic elements such as phage and conjugative plasmids [ 46 ].…”

Section: Discussionmentioning

confidence: 99%

Genetic basis of I-complex plasmid stability and conjugation

et al. 2023

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Plasmids are major drivers of increasing antibiotic resistance, necessitating an urgent need to understand their biology. Here we describe a detailed dissection of the molecular components controlling the genetics of I-complex plasmids, a group of antibiotic resistance plasmids found frequently in pathogenic Escherichia coli and other Enterobacteriaceae that cause significant human disease. We show these plasmids cluster into four distinct subgroups, with the prototype IncI1 plasmid R64 subgroup displaying low nucleotide sequence conservation to other I-complex plasmids. Using pMS7163B, an I-complex plasmid distantly related to R64, we performed a high-resolution transposon-based genetic screen and defined genes involved in replication, stability, and conjugative transfer. We identified the replicon and a partitioning system as essential for replication/stability. Genes required for conjugation included the type IV secretion system, relaxosome, and several uncharacterised genes located in the pMS7163B leading transfer region that exhibited an upstream strand-specific transposon insertion bias. The overexpression of these genes severely impacted host cell growth or reduced fitness during mixed competitive growth, demonstrating that their expression must be controlled to avoid deleterious impacts. These genes were present in >80% of all I-complex plasmids and broadly conserved across multiple plasmid incompatibility groups, implicating an important role in plasmid dissemination.

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Section: Discussionmentioning

confidence: 99%

Genetic basis of I-complex plasmid stability and conjugation

et al. 2023

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“…Each annotated resistant protein was searched against NCBI database using the best match from a blastn search (NCBI BLAST v2.2.27+). Further confirmation was performed by blast search against the Antibiotic Resistance Database (ARDB) (http://ardb.cbcb.umd.edu/) (Balabanov et al, 2012) and Comprehensive Antibiotic Resistance Database (CARD) (https://card.mcmaster.ca/) (McArthur et al, 2013). The NCBI BLAST results were used to confirm the annotation of insertion sequence, replication, and conjugative proteins.…”

Section: Methodsmentioning

confidence: 99%

Characterization of a Novel Conjugative Plasmid in Edwardsiella piscicida Strain MS-18-199

Abdelhamed

Ramachandran

Özdemir

et al. 2019

Front. Cell. Infect. Microbiol.

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Edwardsiella piscicida is a pathogenic bacterium responsible for significant losses in important wild and cultured fish species. E. piscicida strain MS-18-199 recovered from a diseased hybrid catfish from East Mississippi and showed resistance to florfenicol, chloramphenicol, oxytetracycline, doxycycline, erythromycin, tetracycline, azitromycin, spectinomycin, sulfonamide, and bacitracin. To explore the mechanisms of resistance in E. piscicida strain MS-18-199, genomic DNA was extracted and subjected to whole genome sequencing (WGS) using a combination of long (Oxford Nanopore) and short (Illumina) reads. The genome of strain MS-18-199 revealed a novel plasmid named pEPMS-18199. The 117,448 bp plasmid contains several antimicrobial resistance (AMR) elements/genes, including florfenicol efflux pump (floR), tetracycline efflux pump (tetA), tetracycline repressor protein (tetR), sulfonamide resistance (sul2), aminoglycoside O-phosphotransferase aph(6)-Id (strB), and aminoglycoside O-phosphotransferase aph(3)-Ib (strA). Two genes, arsA and arsD, that encode protein components related to transport/resistance to arsenic were also found in pEPMS-18199. In addition, pEPMS-18199 carried twelve conjugative transfer genes (tra), eight transposases and insertion elements, two plasmid stability proteins, two replication proteins, and three partitioning proteins (par system). Results from mobilization and stability experiments revealed that pEPMS-18199 is highly stable in the host cell and could be transferred to Escherichia coli and Edwardsiella ictaluri by conjugation. To our knowledge, this is the first detection of a multidrug resistance (MDR) conjugative plasmid in E. piscicida in the United States. Careful tracking of this plasmid in the aquaculture system is warranted. Knowledge regarding the molecular mechanisms of AMR in aquaculture is important for antimicrobial stewardship.

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“…Ard genes are commonly found in transposons and conjugative plasmids in various prokaryotes [ 57 ]. The Ard antirestriction proteins are of three types, ArdA, ArdB, and ArdC, all of which act as inhibitors of the type I R-M system [ 33 , 34 , 58 , 59 , 60 , 61 ]. ArdA and ArdB proteins are small and acidic, and they both include a small region of similarity that consists of 14 residues designated as the “antirestriction” domain [ 58 ].…”

Section: The Ahpnd-causing Plasmid Pva1mentioning

confidence: 99%

A Review of the Functional Annotations of Important Genes in the AHPND-Causing pVA1 Plasmid

et al. 2020

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Acute hepatopancreatic necrosis disease (AHPND) is a lethal shrimp disease. The pathogenic agent of this disease is a special Vibrio parahaemolyticus strain that contains a pVA1 plasmid. The protein products of two toxin genes in pVA1, pirAvp and pirBvp, targeted the shrimp’s hepatopancreatic cells and were identified as the major virulence factors. However, in addition to pirAvp and pirBvp, pVA1 also contains about ~90 other open-reading frames (ORFs), which may encode functional proteins. NCBI BLASTp annotations of the functional roles of 40 pVA1 genes reveal transposases, conjugation factors, and antirestriction proteins that are involved in horizontal gene transfer, plasmid transmission, and maintenance, as well as components of type II and III secretion systems that may facilitate the toxic effects of pVA1-containing Vibrio spp. There is also evidence of a post-segregational killing (PSK) system that would ensure that only pVA1 plasmid-containing bacteria could survive after segregation. Here, in this review, we assess the functional importance of these pVA1 genes and consider those which might be worthy of further study.

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Comparative analysis of antirestriction activity of the ArdA and ArdB proteins encoded by genes of the R64 transmissible plasmid (IncI1)

Cited by 6 publications

References 15 publications

Genetic basis of I-complex plasmid stability and conjugation

Genetic basis of I-complex plasmid stability and conjugation

Characterization of a Novel Conjugative Plasmid in Edwardsiella piscicida Strain MS-18-199

A Review of the Functional Annotations of Important Genes in the AHPND-Causing pVA1 Plasmid

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