Autonomous Replication of the Conjugative Transposon Tn916
Integrative and conjugative elements (ICEs), also known as conjugative transposons, are self-transferable elements that are widely distributed among bacterial phyla and are important drivers of horizontal gene transfer. Many ICEs carry genes that confer antibiotic resistances to their host cells and...
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| Format: | Article |
| Language: | en_US |
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American Society for Microbiology
2018
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| Online Access: | http://hdl.handle.net/1721.1/117164 https://orcid.org/0000-0003-4999-5486 https://orcid.org/0000-0002-8235-7227 |
| _version_ | 1811090680078925824 |
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| author | Wright, Laurel Grossman, Alan Davis |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Wright, Laurel Grossman, Alan Davis |
| author_sort | Wright, Laurel |
| collection | MIT |
| description | Integrative and conjugative elements (ICEs), also known as conjugative transposons, are self-transferable elements that are widely distributed among bacterial phyla and are important drivers of horizontal gene transfer. Many ICEs carry genes that confer antibiotic resistances to their host cells and are involved in the dissemination of these resistance genes. ICEs reside in host chromosomes but under certain conditions can excise to form a plasmid that is typically the substrate for transfer. A few ICEs are known to undergo autonomous replication following activation. However, it is not clear if autonomous replication is a general property of many ICEs. We found that Tn916, the first conjugative transposon identified, replicates autonomously via a rolling-circle mechanism. Replication of Tn916 was dependent on the relaxase encoded by orf20 of Tn916. The origin of transfer of Tn916, oriT(916), also functioned as an origin of replication. Using immunoprecipitation and mass spectrometry, we found that the relaxase (Orf20) and the two putative helicase processivity factors (Orf22 and Orf23) encoded by Tn916 likely interact in a complex and that the Tn916 relaxase contains a previously unidentified conserved helix-turn-helix domain in its N-terminal region that is required for relaxase function and replication. Lastly, we identified a functional single-strand origin of replication (sso) in Tn916 that we predict primes second-strand synthesis during rolling-circle replication. Together these results add to the emerging data that show that several ICEs replicate via a conserved, rolling-circle mechanism.
IMPORTANCE Integrative and conjugative elements (ICEs) drive horizontal gene transfer and the spread of antibiotic resistances in bacteria. ICEs reside integrated in a host genome but can excise to create a plasmid that is the substrate for transfer to other cells. Here we show that Tn916, an ICE with broad host range, undergoes autonomous rolling-circle replication when in the plasmid form. We found that the origin of transfer functions as a double-stranded origin of replication and identified a single-stranded origin of replication. It was long thought that ICEs do not undergo autonomous replication. Our work adds to the evidence that ICEs replicate autonomously as part of their normal life cycle and indicates that diverse ICEs use the same replicative mechanism. |
| first_indexed | 2024-09-23T14:50:09Z |
| format | Article |
| id | mit-1721.1/117164 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T14:50:09Z |
| publishDate | 2018 |
| publisher | American Society for Microbiology |
| record_format | dspace |
| spelling | mit-1721.1/1171642022-10-01T22:49:27Z Autonomous Replication of the Conjugative Transposon Tn916 Wright, Laurel Grossman, Alan Davis Massachusetts Institute of Technology. Department of Biology Grossman Alan D Wright, Laurel Grossman, Alan Davis Integrative and conjugative elements (ICEs), also known as conjugative transposons, are self-transferable elements that are widely distributed among bacterial phyla and are important drivers of horizontal gene transfer. Many ICEs carry genes that confer antibiotic resistances to their host cells and are involved in the dissemination of these resistance genes. ICEs reside in host chromosomes but under certain conditions can excise to form a plasmid that is typically the substrate for transfer. A few ICEs are known to undergo autonomous replication following activation. However, it is not clear if autonomous replication is a general property of many ICEs. We found that Tn916, the first conjugative transposon identified, replicates autonomously via a rolling-circle mechanism. Replication of Tn916 was dependent on the relaxase encoded by orf20 of Tn916. The origin of transfer of Tn916, oriT(916), also functioned as an origin of replication. Using immunoprecipitation and mass spectrometry, we found that the relaxase (Orf20) and the two putative helicase processivity factors (Orf22 and Orf23) encoded by Tn916 likely interact in a complex and that the Tn916 relaxase contains a previously unidentified conserved helix-turn-helix domain in its N-terminal region that is required for relaxase function and replication. Lastly, we identified a functional single-strand origin of replication (sso) in Tn916 that we predict primes second-strand synthesis during rolling-circle replication. Together these results add to the emerging data that show that several ICEs replicate via a conserved, rolling-circle mechanism. IMPORTANCE Integrative and conjugative elements (ICEs) drive horizontal gene transfer and the spread of antibiotic resistances in bacteria. ICEs reside integrated in a host genome but can excise to create a plasmid that is the substrate for transfer to other cells. Here we show that Tn916, an ICE with broad host range, undergoes autonomous rolling-circle replication when in the plasmid form. We found that the origin of transfer functions as a double-stranded origin of replication and identified a single-stranded origin of replication. It was long thought that ICEs do not undergo autonomous replication. Our work adds to the evidence that ICEs replicate autonomously as part of their normal life cycle and indicates that diverse ICEs use the same replicative mechanism. National Institutes of Health (U.S.) (R01GM050895) National Institute of General Medical Sciences (U.S.) (Grant T32GM007287) 2018-07-27T17:34:39Z 2018-07-27T17:34:39Z 2016-12 Article http://purl.org/eprint/type/JournalArticle 0021-9193 1098-5530 http://hdl.handle.net/1721.1/117164 Wright, Laurel D., and Alan D. Grossman. “Autonomous Replication of the Conjugative Transposon Tn 916.” Journal of Bacteriology, edited by I. B. Zhulin, vol. 198, no. 24, Dec. 2016, pp. 3355–66. https://orcid.org/0000-0003-4999-5486 https://orcid.org/0000-0002-8235-7227 en_US https://doi.org/10.1128/JB.00639-16 Journal of Bacteriology Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf American Society for Microbiology Prof. Grossman via Courtney Crummett |
| spellingShingle | Wright, Laurel Grossman, Alan Davis Autonomous Replication of the Conjugative Transposon Tn916 |
| title | Autonomous Replication of the Conjugative Transposon Tn916 |
| title_full | Autonomous Replication of the Conjugative Transposon Tn916 |
| title_fullStr | Autonomous Replication of the Conjugative Transposon Tn916 |
| title_full_unstemmed | Autonomous Replication of the Conjugative Transposon Tn916 |
| title_short | Autonomous Replication of the Conjugative Transposon Tn916 |
| title_sort | autonomous replication of the conjugative transposon tn916 |
| url | http://hdl.handle.net/1721.1/117164 https://orcid.org/0000-0003-4999-5486 https://orcid.org/0000-0002-8235-7227 |
| work_keys_str_mv | AT wrightlaurel autonomousreplicationoftheconjugativetransposontn916 AT grossmanalandavis autonomousreplicationoftheconjugativetransposontn916 |