DNA Gyrase Inhibitors Increase the Frequency of Bacteriophage-like RcGTA-Mediated Gene Transfer in <i>Rhodobacter capsulatus</i>
<i>Rhodobacter capsulatus</i> produces a bacteriophage-like particle called the gene transfer agent (RcGTA) that mediates horizontal gene transfer. RcGTA particles transfer random ~4.5-kb fragments of genomic DNA that integrate into recipient genomes by allelic replacement. This work add...
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| Format: | Article |
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MDPI AG
2022-11-01
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| Series: | Genes |
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| Online Access: | https://www.mdpi.com/2073-4425/13/11/2071 |
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| author | Rachel Bernelot-Moens J. Thomas Beatty |
| author_facet | Rachel Bernelot-Moens J. Thomas Beatty |
| author_sort | Rachel Bernelot-Moens |
| collection | DOAJ |
| description | <i>Rhodobacter capsulatus</i> produces a bacteriophage-like particle called the gene transfer agent (RcGTA) that mediates horizontal gene transfer. RcGTA particles transfer random ~4.5-kb fragments of genomic DNA that integrate into recipient genomes by allelic replacement. This work addresses the effect of sub-inhibitory concentrations of antibiotics on gene transfer by RcGTA. A transduction assay was developed to test the effects of various substances on gene transfer. Using this assay, low concentrations of DNA gyrase inhibitors were found to increase the frequency of gene transfer. Novobiocin was studied in more detail, and it was found that this antibiotic did not influence the production or release of RcGTA but instead appeared to act on the recipient cells. The target of novobiocin in other species has been shown to be the GyrB subunit of DNA gyrase (a heterotetramer of 2GyrA and 2GyrB). <i>R. capsulatus</i> encodes GyrA and GyrB homologues, and a GyrB overexpression plasmid was created and found to confer resistance to novobiocin. The presence of the overexpression plasmid in recipient cells greatly diminished the novobiocin-mediated increase in gene transfer, confirming that this effect is due to the binding of novobiocin by GyrB. The results of this work show that antibiotics affect gene transfer in <i>R. capsulatus</i> and may be relevant to microbial genetic exchange in natural ecosystems. |
| first_indexed | 2024-03-09T19:03:16Z |
| format | Article |
| id | doaj.art-2daef8994d6945b98fdc5b390d2d8b43 |
| institution | Directory Open Access Journal |
| issn | 2073-4425 |
| language | English |
| last_indexed | 2024-03-09T19:03:16Z |
| publishDate | 2022-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Genes |
| spelling | doaj.art-2daef8994d6945b98fdc5b390d2d8b432023-11-24T04:49:19ZengMDPI AGGenes2073-44252022-11-011311207110.3390/genes13112071DNA Gyrase Inhibitors Increase the Frequency of Bacteriophage-like RcGTA-Mediated Gene Transfer in <i>Rhodobacter capsulatus</i>Rachel Bernelot-Moens0J. Thomas Beatty1Department of Microbiology and Immunology, The University of British Columbia, 2350 Health Sciences Mall, Vancouver, BC V6T 1Z3, CanadaDepartment of Microbiology and Immunology, The University of British Columbia, 2350 Health Sciences Mall, Vancouver, BC V6T 1Z3, Canada<i>Rhodobacter capsulatus</i> produces a bacteriophage-like particle called the gene transfer agent (RcGTA) that mediates horizontal gene transfer. RcGTA particles transfer random ~4.5-kb fragments of genomic DNA that integrate into recipient genomes by allelic replacement. This work addresses the effect of sub-inhibitory concentrations of antibiotics on gene transfer by RcGTA. A transduction assay was developed to test the effects of various substances on gene transfer. Using this assay, low concentrations of DNA gyrase inhibitors were found to increase the frequency of gene transfer. Novobiocin was studied in more detail, and it was found that this antibiotic did not influence the production or release of RcGTA but instead appeared to act on the recipient cells. The target of novobiocin in other species has been shown to be the GyrB subunit of DNA gyrase (a heterotetramer of 2GyrA and 2GyrB). <i>R. capsulatus</i> encodes GyrA and GyrB homologues, and a GyrB overexpression plasmid was created and found to confer resistance to novobiocin. The presence of the overexpression plasmid in recipient cells greatly diminished the novobiocin-mediated increase in gene transfer, confirming that this effect is due to the binding of novobiocin by GyrB. The results of this work show that antibiotics affect gene transfer in <i>R. capsulatus</i> and may be relevant to microbial genetic exchange in natural ecosystems.https://www.mdpi.com/2073-4425/13/11/2071<i>Rhodobacter</i>gene transfer agentRcGTAhorizontal gene transferphagevirus |
| spellingShingle | Rachel Bernelot-Moens J. Thomas Beatty DNA Gyrase Inhibitors Increase the Frequency of Bacteriophage-like RcGTA-Mediated Gene Transfer in <i>Rhodobacter capsulatus</i> Genes <i>Rhodobacter</i> gene transfer agent RcGTA horizontal gene transfer phage virus |
| title | DNA Gyrase Inhibitors Increase the Frequency of Bacteriophage-like RcGTA-Mediated Gene Transfer in <i>Rhodobacter capsulatus</i> |
| title_full | DNA Gyrase Inhibitors Increase the Frequency of Bacteriophage-like RcGTA-Mediated Gene Transfer in <i>Rhodobacter capsulatus</i> |
| title_fullStr | DNA Gyrase Inhibitors Increase the Frequency of Bacteriophage-like RcGTA-Mediated Gene Transfer in <i>Rhodobacter capsulatus</i> |
| title_full_unstemmed | DNA Gyrase Inhibitors Increase the Frequency of Bacteriophage-like RcGTA-Mediated Gene Transfer in <i>Rhodobacter capsulatus</i> |
| title_short | DNA Gyrase Inhibitors Increase the Frequency of Bacteriophage-like RcGTA-Mediated Gene Transfer in <i>Rhodobacter capsulatus</i> |
| title_sort | dna gyrase inhibitors increase the frequency of bacteriophage like rcgta mediated gene transfer in i rhodobacter capsulatus i |
| topic | <i>Rhodobacter</i> gene transfer agent RcGTA horizontal gene transfer phage virus |
| url | https://www.mdpi.com/2073-4425/13/11/2071 |
| work_keys_str_mv | AT rachelbernelotmoens dnagyraseinhibitorsincreasethefrequencyofbacteriophagelikercgtamediatedgenetransferinirhodobactercapsulatusi AT jthomasbeatty dnagyraseinhibitorsincreasethefrequencyofbacteriophagelikercgtamediatedgenetransferinirhodobactercapsulatusi |