Intracellular Transposition of Mobile Genetic Elements Associated with the Colistin Resistance Gene mcr-1
ABSTRACT Mobile colistin resistance (mcr) genes are often located on conjugative plasmids, where their association with insertion sequences enables intercellular and intracellular dissemination throughout bacterial replicons and populations. Multiple mcr genes have been discovered in every habitable...
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2023-02-01
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| Series: | Microbiology Spectrum |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.03278-22 |
| _version_ | 1828033535507693568 |
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| author | Richard N. Goodman Supathep Tansirichaiya Michael S. M. Brouwer Adam P. Roberts |
| author_facet | Richard N. Goodman Supathep Tansirichaiya Michael S. M. Brouwer Adam P. Roberts |
| author_sort | Richard N. Goodman |
| collection | DOAJ |
| description | ABSTRACT Mobile colistin resistance (mcr) genes are often located on conjugative plasmids, where their association with insertion sequences enables intercellular and intracellular dissemination throughout bacterial replicons and populations. Multiple mcr genes have been discovered in every habitable continent, in many bacterial species, on both plasmids and integrated into the chromosome. Previously, we showed the intercellular transfer of mcr-1 on an IncI1 plasmid, pMCR-E2899, between strains of Escherichia coli. Characterizing the intracellular dynamics of mcr-1 transposition and recombination would further our understanding of how these important genes move through bacterial populations and whether interventions can be put in place to stop their spread. In this study, we aimed to characterize transfer events from the mcr-1-containing transposon Tn7511 (ISApl1-mcr-1-pap2-ISApl1), located on plasmid pMCR-E2899, using the pBACpAK entrapment vector. Following the transformation of pBACpAK into our DH5α-Azir/pMCR-E2899 transconjugant, we captured ISApl1 in pBACpAK multiple times and, for the first time, observed the ISApl1-mediated transfer of the mcr-1 transposon (Tn7511) into the chromosome of E. coli DH5α. Whole-genome sequencing allowed us to determine consensus insertion sites of ISApl1 and Tn7511 in this strain, and comparison of these sites allowed us to explain the transposition events observed. These observations reveal the consequences of ISApl1 transposition within and between multiple replicons of the same cell and show mcr-1 transposition within the cell as part of the novel transposon Tn7511. IMPORTANCE By analyzing the intracellular transfer of clinically relevant transposons, we can understand the dissemination and evolution of drug resistance conferring mobile genetic elements (MGEs) once a plasmid enters a cell following conjugation. This knowledge will help further our understanding of how these important genes move through bacterial populations. Utilizing the pBACpAK entrapment vector has allowed us to determine the mobility of the novel mcr-1-containing transposon Tn7511. |
| first_indexed | 2024-04-10T15:21:06Z |
| format | Article |
| id | doaj.art-00c92aeb3b664f329d4af0a6a5bc911a |
| institution | Directory Open Access Journal |
| issn | 2165-0497 |
| language | English |
| last_indexed | 2024-04-10T15:21:06Z |
| publishDate | 2023-02-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj.art-00c92aeb3b664f329d4af0a6a5bc911a2023-02-14T14:15:50ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972023-02-0111110.1128/spectrum.03278-22Intracellular Transposition of Mobile Genetic Elements Associated with the Colistin Resistance Gene mcr-1Richard N. Goodman0Supathep Tansirichaiya1Michael S. M. Brouwer2Adam P. Roberts3Department of Tropical Disease Biology, Liverpool School of Tropical Medicine, Liverpool, United KingdomDepartment of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, ThailandWageningen Bioveterinary Research, Lelystad, The NetherlandsDepartment of Tropical Disease Biology, Liverpool School of Tropical Medicine, Liverpool, United KingdomABSTRACT Mobile colistin resistance (mcr) genes are often located on conjugative plasmids, where their association with insertion sequences enables intercellular and intracellular dissemination throughout bacterial replicons and populations. Multiple mcr genes have been discovered in every habitable continent, in many bacterial species, on both plasmids and integrated into the chromosome. Previously, we showed the intercellular transfer of mcr-1 on an IncI1 plasmid, pMCR-E2899, between strains of Escherichia coli. Characterizing the intracellular dynamics of mcr-1 transposition and recombination would further our understanding of how these important genes move through bacterial populations and whether interventions can be put in place to stop their spread. In this study, we aimed to characterize transfer events from the mcr-1-containing transposon Tn7511 (ISApl1-mcr-1-pap2-ISApl1), located on plasmid pMCR-E2899, using the pBACpAK entrapment vector. Following the transformation of pBACpAK into our DH5α-Azir/pMCR-E2899 transconjugant, we captured ISApl1 in pBACpAK multiple times and, for the first time, observed the ISApl1-mediated transfer of the mcr-1 transposon (Tn7511) into the chromosome of E. coli DH5α. Whole-genome sequencing allowed us to determine consensus insertion sites of ISApl1 and Tn7511 in this strain, and comparison of these sites allowed us to explain the transposition events observed. These observations reveal the consequences of ISApl1 transposition within and between multiple replicons of the same cell and show mcr-1 transposition within the cell as part of the novel transposon Tn7511. IMPORTANCE By analyzing the intracellular transfer of clinically relevant transposons, we can understand the dissemination and evolution of drug resistance conferring mobile genetic elements (MGEs) once a plasmid enters a cell following conjugation. This knowledge will help further our understanding of how these important genes move through bacterial populations. Utilizing the pBACpAK entrapment vector has allowed us to determine the mobility of the novel mcr-1-containing transposon Tn7511.https://journals.asm.org/doi/10.1128/spectrum.03278-22entrapment vectorinsertion sequencetarget siteantimicrobial resistanceplasmidISApl1 |
| spellingShingle | Richard N. Goodman Supathep Tansirichaiya Michael S. M. Brouwer Adam P. Roberts Intracellular Transposition of Mobile Genetic Elements Associated with the Colistin Resistance Gene mcr-1 Microbiology Spectrum entrapment vector insertion sequence target site antimicrobial resistance plasmid ISApl1 |
| title | Intracellular Transposition of Mobile Genetic Elements Associated with the Colistin Resistance Gene mcr-1 |
| title_full | Intracellular Transposition of Mobile Genetic Elements Associated with the Colistin Resistance Gene mcr-1 |
| title_fullStr | Intracellular Transposition of Mobile Genetic Elements Associated with the Colistin Resistance Gene mcr-1 |
| title_full_unstemmed | Intracellular Transposition of Mobile Genetic Elements Associated with the Colistin Resistance Gene mcr-1 |
| title_short | Intracellular Transposition of Mobile Genetic Elements Associated with the Colistin Resistance Gene mcr-1 |
| title_sort | intracellular transposition of mobile genetic elements associated with the colistin resistance gene mcr 1 |
| topic | entrapment vector insertion sequence target site antimicrobial resistance plasmid ISApl1 |
| url | https://journals.asm.org/doi/10.1128/spectrum.03278-22 |
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