Abstract
Legionella pneumophila is an environmental bacterium and the causative agent of Legionnaires’ disease. Previous genomic studies have shown that recombination accounts for a high proportion (>96%) of diversity within several major disease-associated sequence types (STs) of L. pneumophila. This suggests that recombination represents a potentially important force shaping adaptation and virulence. Despite this, little is known about the biological effects of recombination in L. pneumophila, particularly with regards to homologous recombination (whereby genes are replaced with alternative allelic variants). Using newly available population genomic data, we have disentangled events arising from homologous and non-homologous recombination in six major disease-associated STs of L. pneumophila (subsp. pneumophila), and subsequently performed a detailed characterisation of the dynamics and impact of homologous recombination. We identified genomic “hotspots” of homologous recombination that include regions containing outer membrane proteins, the lipopolysaccharide (LPS) region and Dot/Icm effectors, which provide interesting clues to the selection pressures faced by L. pneumophila. Inference of the origin of the recombined regions showed that isolates have most frequently imported DNA from isolates belonging to their own clade, but also occasionally from other major clades of the same subspecies. This supports the hypothesis that the possibility for horizontal exchange of new adaptations between major clades of the subspecies may have been a critical factor in the recent emergence of several clinically important STs from diverse genomic backgrounds. However, acquisition of recombined regions from another subspecies, L. pneumophila subsp. fraseri, was rarely observed, suggesting the existence of a recombination barrier and/or the possibility of ongoing speciation between the two subspecies. Finally, we suggest that multi-fragment recombination may occur in L. pneumophila, whereby multiple non-contiguous segments that originate from the same molecule of donor DNA are imported into a recipient genome during a single episode of recombination.
Highlights
While all bacteria reproduce clonally, some import DNA from other organisms into their chromosomes through processes such as recombination or horizontal gene transfer
Previous studies have shown that members of this species undergo a process called recombination, whereby DNA is imported from another bacterial cell into the recipient genome
Whilst recombination plays an undoubtedly important role in L. pneumophila evolution, accounting for more than 96% of the diversity observed within some lineages, little is known about its biological impact
Summary
While all bacteria reproduce clonally, some import DNA from other organisms into their chromosomes through processes such as recombination or horizontal gene transfer. The imported DNA can either replace a homologous segment of the genome (homologous recombination) or comprise novel genes that are new to the recipient genome (non-homologous recombination) The former results in the replacement of genes with alternative allelic variants and requires the DNA to be highly similar, and possibly identical, at both ends of the fragment [1]. The importance of recombination in bacterial evolution first became clear through the analysis of multi-locus sequence typing (MLST) data, which showed that phylogenetic trees constructed from individual MLST genes were often incongruent [2] These analyses predicted that the rate of homologous recombination varies considerably between different species [3]. A recent study has suggested that bacteria use recombination to delete selfish mobile genetic elements from their genomes [8]
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