Experimental capture of genomic islands defines a widespread class of genetic element capable of non-autonomous transfer

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Abstract

Bacteria acquire new genes by horizontal gene transfer (HGT). Acquisition is typically mediated by mobile genetic elements (MGEs), however, beyond plasmids, bacteriophages and certain integrative conjugative elements (ICEs), the nature and diversity of MGEs is poorly understood. The bacterium Pseudomonas fluorescens SBW25 was propagated by serial transfer in the presence of filtrate obtained from garden compost communities. Genome sequencing of derived colonies revealed acquisition of three different mobile elements, each integrated immediately downstream of tmRNA . All are flanked by direct repeats and harbour a tyrosine integrase ( intY ), followed by a cargo of accessory genes including putative phage defence systems. Although characteristic of genomic islands, MGE-classifiers showed no matches to mobile elements. Interrogation of DNA sequence databases showed that similar elements are widespread in the genus Pseudomonas and beyond, with Vibrio Pathogenicity Island-1 (VPI-1) from V. cholerae being a notable example. Bioinformatic analyses demonstrate frequent transfer among diverse hosts. With focus on a single 55 kb element (I55) we show that intY is necessary for excision and circularisation, that the element is incapable of autonomous horizontal transfer, but is mobilizable – in the absence of direct cell-cell contact – upon addition of community filtrate. Further analyses demonstrate that I55 enhances host fitness in the presence of community filtrate, which stems in part from ability to defend against phages.

Significance Statement

The impact of horizontal gene transfer on the evolution of bacteria outpaces that driven by spontaneous mutation, but knowledge of the range of mediators, the genes mobilised, and the mechanisms of movement have largely depended on inferences stemming from bioinformatics. Here we describe a real-time evolution experiment in which a single focal strain propagated with filtrate from a complex microbial community captures genetic elements carrying a diverse cargo of genes whose mobility was previously unknown. The elements are representative of a class of mobile DNA that depend on nothing other than a tyrosine integrase that targets a highly conserved gene. The elements encode phage resistance, require the integrase to excise from the genome, confer a significant fitness advantage, and depend on a secondary element for transfer.

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