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bioRxiv · 10.1101/2022.01.07.475358

Homologous recombination between tandem paralogues drives evolution of Type VII secretion system immunity genes in firmicute bacteria

Garrett, S.·Mariano, G.·Palmer, T.
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Abstract

The Type VII secretion system (T7SS) is found in many Gram-positive firmicutes and secretes protein toxins that mediate bacterial antagonism. Two T7SS toxins have been identified in Staphylococcus aureus, EsaD a nuclease toxin that is counteracted by the EsaG immunity protein, and TspA, which has membrane depolarising activity and is neutralised by TsaI. Both toxins are polymorphic, and strings of non-identical esaG and tsaI immunity genes are encoded in all S. aureus strains. To investigate the evolution of esaG repertoires, we analysed the sequences of the tandem esaG genes and their encoded proteins. We identified three blocks of high sequence similarity shared by all esaG genes and identified evidence of extensive recombination events between esaG paralogues facilitated through these conserved sequence blocks. Recombination between these blocks accounts for loss and expansion of esaG genes in S. aureus genomes and we identified evidence of such events during evolution of strains in clonal complex 8. TipC, an immunity protein for the TelC lipid II phosphatase toxin secreted by the streptococcal T7SS, is also encoded by multiple gene paralogues. Two blocks of high sequence similarity locate to the 5 and 3 end of tipC genes, and we found strong evidence for recombination between tipC paralogues encoded by Streptococcus mitis BCC08. By contrast, we found only a single homology block across tsaI genes, and little evidence for intergenic recombination within this gene family. We conclude that homologous recombination is one of the drivers for the evolution of T7SS immunity gene clusters. DATA SUMMARYAll sequence data for strains used in this study are available on NCBI under BioProject PRJNA789916. Sequences from the NCTC3000 project are available on NCBI under BioProject PRJEB6403. Supplementary data 2 and all custom scripts are available on Github: https://github.com/GM110Z/Garret-et-al.-recombination-paper. IMPACT STATEMENTThe type VII secretion system (T7SS) in firmicutes secretes polymorphic toxins that target other bacteria. To protect from the action of these toxins, bacteria carry multiple paralogous copies of immunity protein-encoding genes that are sequence-related but non-identical. To date, little is known about how T7 immunity gene families evolve. In this study we analysed a cluster of EsaG-encoding genes in Staphylococcus aureus which are found at the ess/T7 secretion locus and provide immunity against the T7 secreted nuclease toxin, EsaD. We identified three homology blocks covering esaG genes and their downstream intergenic regions, which are separated by two variable regions. We have shown that recombination can occur between these homology blocks, leading to loss or expansion of esaG genes at this locus. Using a historical dataset of closely related S. aureus strains from clonal complex 8, we identified several independent recombination events leading to changes in the esaG repertoire. We further showed that similar events are observed for an immunity protein encoded by Group B Streptococcus spp. suggesting that recombination plays a broader role in the evolution of T7SS immunity-encoding genes. We speculate that gain and loss of T7 immunity genes is weighed in response to environmental pressure and metabolic burden.

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Garrett, S., Mariano, G., Palmer, T.. 2022-01-08. Homologous recombination between tandem paralogues drives evolution of Type VII secretion system immunity genes in firmicute bacteria. https://doi.org/10.1101/2022.01.07.475358

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