Extensive Cotransformation of Natural Variation into Chromosomes of Naturally Competent Haemophilus influenzae

Author:

Mell Joshua Chang11,Lee Jae Yun2,Firme Marlo2,Sinha Sunita3,Redfield Rosemary J1

Affiliation:

1. Department of Zoology, University of British Columbia, Vancouver, BC V6T 1Z3, Canada

2. Genome Sciences and Technology Graduate Program, University of British Columbia, Vancouver, BC V6T 1Z3, Canada

3. Department of Pharmacy Sciences, University of British Columbia, Vancouver, BC V6T 1Z3, Canada

Abstract

Abstract Naturally competent bacterial species actively take up environmental DNA and can incorporate it into their chromosomes by homologous recombination. This can bring genetic variation from environmental DNA to recipient chromosomes, often in multiple long “donor” segments. Here, we report the results of genome sequencing 96 colonies of a laboratory Haemophilus influenzae strain, which had been experimentally transformed by DNA from a diverged clinical isolate. Donor segments averaged 6.9 kb (spanning several genes) and were clustered into recombination tracts of ~19.5 kb. Individual colonies had replaced from 0.1 to 3.2% of their chromosomes, and ~1/3 of all donor-specific single-nucleotide variants were present in at least one recombinant. We found that nucleotide divergence did not obviously limit the locations of recombination tracts, although there were small but significant reductions in divergence at recombination breakpoints. Although indels occasionally transformed as parts of longer recombination tracts, they were common at breakpoints, suggesting that indels typically block progression of strand exchange. Some colonies had recombination tracts in which variant positions contained mixtures of both donor and recipient alleles. These tracts were clustered around the origin of replication and were interpreted as the result of heteroduplex segregation in the original transformed cell. Finally, a pilot experiment demonstrated the utility of natural transformation for genetically dissecting natural phenotypic variation. We discuss our results in the context of the potential to merge experimental and population genetic approaches, giving a more holistic understanding of bacterial gene transfer.

Publisher

Oxford University Press (OUP)

Subject

Genetics(clinical),Genetics,Molecular Biology

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