Cryo-EM structure and evolutionary history of the conjugation surface exclusion protein TraT

Chloe Seddon, Sophia David, Joshua L. C. Wong, Naito Ishimoto, Shan He, Jonathan Bradshaw, Wen Wen Low, Gad Frankel () and Konstantinos Beis ()
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Chloe Seddon: Imperial College London
Sophia David: University of Oxford
Joshua L. C. Wong: Imperial College London
Naito Ishimoto: Imperial College London
Shan He: Imperial College London
Jonathan Bradshaw: Imperial College London
Wen Wen Low: Imperial College London
Gad Frankel: Imperial College London
Konstantinos Beis: Imperial College London

Nature Communications, 2025, vol. 16, issue 1, 1-15

Abstract: Abstract Conjugation plays a major role in dissemination of antimicrobial resistance genes. Following transfer of IncF-like plasmids, recipients become refractory to a second wave of conjugation with the same plasmid via entry (TraS) and surface (TraT) exclusion mechanisms. Here, we show that TraT from the pKpQIL and F plasmids (TraTpKpQIL and TraTF) exhibits plasmid surface exclusion specificity. The cryo-EM structures of TraTpKpQIL and TraTF reveal that they oligomerise into decameric champagne bottle cork-like structures, which are anchored to the outer membrane via a diacylglycerol and palmitic acid modified α-helical barrel domain. Unexpectedly, we identify chromosomal TraT homologues from multiple Gram-negative phyla which form numerous divergent lineages in a phylogenetic tree of TraT sequences. Plasmid-associated TraT sequences are found in multiple distinct lineages, including two separate clades incorporating TraT from Enterobacteriaceae IncF/F-like and Legionellaceae F-like plasmids. These findings suggest that different plasmid backbones have acquired and co-opted TraT on independent occasions.

Date: 2025
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DOI: 10.1038/s41467-025-55834-w

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