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dc.contributor.authorBowring, Janine Zara
dc.contributor.authorSu, Yue
dc.contributor.authorAlsaadi, Ahlam
dc.contributor.authorSvenningsen, Sine L
dc.contributor.authorParkhill, Julian
dc.contributor.authorIngmer, Hanne
dc.date.accessioned2022-03-14T02:04:59Z
dc.date.available2022-03-14T02:04:59Z
dc.date.issued2022-02-23
dc.identifier.issn2165-0497
dc.identifier.otherPMC8826898
dc.identifier.other35138167
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/334953
dc.description.abstractBacteriophage-mediated transduction of bacterial DNA is a major route of horizontal gene transfer in the human pathogen, Staphylococcus aureus. Transduction involves the packaging of bacterial DNA by viruses and enables the transmission of virulence and resistance genes between cells. To learn more about transduction in S. aureus, we searched a transposon mutant library for genes and mutations that enhanced transfer mediated by the temperate phage, ϕ11. Using a novel screening strategy, we performed multiple rounds of transduction of transposon mutant pools selecting for an antibiotic resistance marker within the transposon element. When determining the locations of transferred mutations, we found that the screen had selected for just 1 or 2 transposon mutant(s) within each pool of 96 mutants. Subsequent analysis showed that the position of the transposon, rather than the inactivation of bacterial genes, was responsible for the phenotype. Interestingly, from multiple rounds, we identified a pattern of transduction that encompassed mobile genetic elements as well as chromosomal regions both upstream and downstream of the phage integration site. The latter was confirmed by DNA sequencing of purified phage lysates. Importantly, transduction frequencies were lower for phage lysates obtained by phage infection rather than induction. Our results confirmed previous reports of lateral transduction of bacterial DNA downstream of the integrated phage but also indicated a novel form of specialized transduction of DNA upstream of the phage. These findings illustrated the complexity of transduction processes and increased our understanding of the mechanisms by which phages transfer bacterial DNA. IMPORTANCE Horizontal transfer of DNA between bacterial cells contributes to the spread of virulence and antibiotic resistance genes in human pathogens. For Staphylococcus aureus, bacterial viruses play a major role in facilitating the horizontal transfer. These viruses, termed bacteriophages, can transfer bacterial DNA between cells by a process known as transduction, which despite its importance is only poorly characterized. Here, we employed a transposon mutant library to investigate transduction in S. aureus. We showed that the genomic location of bacterial DNA relative to where bacteriophages integrated into that bacterial genome affected how frequently that DNA was transduced. Based on serial transduction of transposon mutant pools and direct sequencing of bacterial DNA in bacteriophage particles, we demonstrated both lateral and specialized transduction. The use of mutant libraries to investigate the genomic patterns of bacterial DNA transferred between cells could help us understand how horizontal transfer influences virulence and resistance development.
dc.languageeng
dc.publisherAmerican Society for Microbiology
dc.rightsAttribution 4.0 International
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.sourcenlmid: 101634614
dc.sourceessn: 2165-0497
dc.subjectPhages
dc.subjectTransduction
dc.subjectStaphylococcus aureus
dc.subjectGene transfer
dc.subjectBacteriophages
dc.subjectTransposons
dc.subjectMobile genetic elements
dc.subjectTransducing Particles
dc.titleScreening for Highly Transduced Genes in Staphylococcus aureus Revealed Both Lateral and Specialized Transduction.
dc.typeArticle
dc.date.updated2022-03-14T02:04:58Z
prism.issueIdentifier1
prism.publicationNameMicrobiol Spectr
prism.volume10
dc.identifier.doi10.17863/CAM.82391
rioxxterms.versionofrecord10.1128/spectrum.02423-21
rioxxterms.versionVoR
rioxxterms.licenseref.urihttps://creativecommons.org/licenses/by/4.0/
dc.contributor.orcidBowring, Janine Zara [0000-0003-2448-9632]
dc.contributor.orcidParkhill, Julian [0000-0002-7069-5958]
dc.contributor.orcidIngmer, Hanne [0000-0002-8350-5631]
dc.identifier.eissn2165-0497
pubs.funder-project-idTeknologi og Produktion, Det Frie Forskningsråd (7017-00079B)
pubs.funder-project-idWellcome Trust (WT098051)
pubs.funder-project-idOlav Thon Foundation (Development of antibiotic resistance in bacterial biofilms)
cam.issuedOnline2022-02-09


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Attribution 4.0 International
Except where otherwise noted, this item's licence is described as Attribution 4.0 International