Abstract

Genetic exchange mediated by viruses of bacteria (bacteriophages) is the primary driver of rapid bacterial evolution. The priority of viruses is usually to propagate themselves. Most bacteriophages use the small terminase protein to identify their own genome and direct its inclusion into phage capsids. Gene transfer agents (GTAs) are descended from bacteriophages, but they instead package fragments of the entire bacterial genome without preference for their own genes. GTAs do not selectively target specific DNA, and no GTA small terminases are known. Here, we identified the small terminase from the model Rhodobacter capsulatus GTA, which then allowed prediction of analogues in other species. We examined the role of the small terminase in GTA production and propose a structural basis for random DNA packaging.IMPORTANCE Random transfer of any and all genes between bacteria could be influential in the spread of virulence or antimicrobial resistance genes. Discovery of the true prevalence of GTAs in sequenced genomes is hampered by their apparent similarity to bacteriophages. Our data allowed the prediction of small terminases in diverse GTA producer species, and defining the characteristics of a "GTA-type" terminase could be an important step toward novel GTA identification. Importantly, the GTA small terminase shares many features with its phage counterpart. We propose that the GTA terminase complex could become a streamlined model system to answer fundamental questions about double-stranded DNA (dsDNA) packaging by viruses that have not been forthcoming to date.

Highlights

  • Genetic exchange mediated by viruses of bacteria is the primary driver of rapid bacterial evolution

  • Viral transduction by bacteriophages is generally accepted to be the dominant mechanism for the rapid exchange of genes between bacteria

  • We definitively identify the small terminase of the model R. capsulatus Gene transfer agents (GTAs), demonstrate and localize its interaction with the large terminase, and investigate its role in RcGTA production

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Summary

Introduction

Genetic exchange mediated by viruses of bacteria (bacteriophages) is the primary driver of rapid bacterial evolution. Most bacteriophages use the small terminase protein to identify their own genome and direct its inclusion into phage capsids. Gene transfer agents (GTAs) are descended from bacteriophages, but they instead package fragments of the entire bacterial genome without preference for their own genes. Gene transfer agents (GTAs) are small virus-like particles that exclusively package and transfer random fragments of their host bacterium’s DNA to recipient bacteria [7, 8], with no preference for the propagation of December 2019 Volume 93 Issue 23 e01328-19. An eye-opening study of antibiotic gene transfer by GTAs in in situ marine microcosms detected extraordinary transfer frequencies that were orders of magnitude greater than more established mechanisms [11]

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