Mobilization and prevalence of a fusobacterial plasmid

Claypool, Brianna M.; Yoder, Sean C.; Citron, Diane M.; Finegold, Sydney M.; Goldstein, Ellie J. C.; Haake, Susan Kinder

doi:10.1016/j.plasmid.2009.09.001

Cited by 7 publications

(3 citation statements)

References 34 publications

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“…Several plasmids of F. nucleatum , including pFN1, have relaxase gene homologs that may operate in plasmid mobilization. Recently, Claypool et al (41) found that the fusobacterial plasmid pFN1 encodes genes, enabling efficient mobilization between strains of Escherichia coli by the broad-range plasmid RP4. A role was suggested for plasmid mobilization in DNA transfer in F. nucleatum .…”

Section: Dna Transfer In Major Dental Plaque Bacteriamentioning

confidence: 99%

“…A role was suggested for plasmid mobilization in DNA transfer in F. nucleatum . Plasmid or plasmid-related sequences were identified in as many as 11.5% of the strains in a panel of clinical F. nucleatum isolates (41). Roberts and Lansciardi (42) showed that two strains of F. nucleatum were able to transfer tetracycline resistance by conjugation encoded by the tet (M) determinant to strains of F. nucleatum , Peptostreptococcus anaerobius , and Enterococcus faecalis .…”

Section: Dna Transfer In Major Dental Plaque Bacteriamentioning

confidence: 99%

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Bacterial sex in dental plaque

Olsen

Tribble

Fiehn

et al. 2013

Journal of Oral Microbiology

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Genes are transferred between bacteria in dental plaque by transduction, conjugation, and transformation. Membrane vesicles can also provide a mechanism for horizontal gene transfer. DNA transfer is considered bacterial sex, but the transfer is not parallel to processes that we associate with sex in higher organisms. Several examples of bacterial gene transfer in the oral cavity are given in this review. How frequently this occurs in dental plaque is not clear, but evidence suggests that it affects a number of the major genera present. It has been estimated that new sequences in genomes established through horizontal gene transfer can constitute up to 30% of bacterial genomes. Gene transfer can be both inter- and intrageneric, and it can also affect transient organisms. The transferred DNA can be integrated or recombined in the recipient's chromosome or remain as an extrachromosomal inheritable element. This can make dental plaque a reservoir for antimicrobial resistance genes. The ability to transfer DNA is important for bacteria, making them better adapted to the harsh environment of the human mouth, and promoting their survival, virulence, and pathogenicity.

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Section: Dna Transfer In Major Dental Plaque Bacteriamentioning

confidence: 99%

Section: Dna Transfer In Major Dental Plaque Bacteriamentioning

confidence: 99%

Bacterial sex in dental plaque

Olsen

Tribble

Fiehn

et al. 2013

Journal of Oral Microbiology

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“…polymorphum (Fnp 10953; transformation by electroporation) 52 , Fusobacterium nucleatum subsp. polymorphum 12230 ( Fnp 12230; transformation by sonoporation) 53 , and a recent paper highlighting the first gene interruption in Fusobacterium necrophorum using DNA conjugation from E. coli 54,55 . Needless to say, these four strains do not encompass all of the Fusobacterium subspecies and their respective infections and diseases that we would like to study and highlights the need for molecular biology and biochemical studies to achieve universal genetics.…”

Section: Introductionmentioning

confidence: 99%

DNA methyltransferase enhanced Fusobacterium nucleatum genetics

Umaña

Nguyen

Sanders

et al. 2022

Preprint

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Bacterial restriction-modification (R-M) systems are a first line immune defense against foreign DNA from viruses and other bacteria. While R-M systems are critical in maintaining genome integrity, R-M nucleases unfortunately present significant barriers to targeted genetic modification. Bacteria of the genus Fusobacterium are oral, Gram-negative, anaerobic, opportunistic pathogens that are implicated in the progression and severity of multiple cancers and tissue infections, yet our understanding of their direct roles in disease have been severely hindered by their genetic recalcitrance. Here, we demonstrate a path to overcome these barriers in Fusobacterium by using native DNA methylation as a host mimicry strategy to bypass R-M system cleavage of user introduced plasmid DNA. We report the identification, characterization, and successful use of Fusobacterium nucleatum (Fn) Type II and III DNA methyltransferase (DMTase) enzymes to produce a multi-fold increase in gene knockout efficiency in the strain Fusobacterium nucleatum subsp. nucleatum 23726 (Fnn 23726), as well as the first efficient gene knockouts and complementations in Fnn 25586. We show plasmid protection can be accomplished in vitro with purified enzymes, as well as in vivo in an E. coli host that constitutively expresses Fnn DMTase enzymes. By characterizing specific DMTases that are critical for bypassing R-M systems, we have enhanced our understanding of potential enzyme combinations, with the goal of expanding these studies to genetically modify clinical isolates of Fusobacterium that have thus far been inaccessible to molecular characterization. This proof-of-concept study provides a roadmap to guide molecular microbiology efforts of the scientific community to facilitate the discovery of new Fusobacterium virulence genes, thereby leading to a new era of characterizing how an oral opportunistic pathogen contributes to an array of human infections and diseases.IMPORTANCEFusobacterium nucleatum is an oral opportunistic pathogen associated with diseases including cancer and preterm birth. Our understanding of how this bacterium modulates human disease has been hindered by a lack of genetic systems. Here we show that F. nucleatum DNA methyltransferase modified plasmid DNA overcomes the transformation barrier and allows the development of genetic systems in previously inaccessible strains. We present a strategy that can be expanded to enable the genetic modification of clinical isolates, thereby fostering investigational studies to uncover novel host-pathogen interactions in Fusobacterium.

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