Structure and rational engineering of the PglX methyltransferase and specificity factor for BREX phage defence.

Autor: Went SC; Department of Biosciences, Durham University, South Road, Durham, UK., Picton DM; Department of Biosciences, Durham University, South Road, Durham, UK., Morgan RD; New England Biolabs, 240 County Road, Ipswich, MA, USA., Nelson A; Faculty of Health and Life Sciences, Northumbria University, Newcastle Upon Tyne, UK., Brady A; Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, Liverpool, UK., Mariano G; Department of Microbial Sciences, Faculty of Health and Medical Sciences, University of Surrey, Guildford, UK., Dryden DTF; Department of Biosciences, Durham University, South Road, Durham, UK., Smith DL; Faculty of Health and Life Sciences, Northumbria University, Newcastle Upon Tyne, UK., Wenner N; Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, Liverpool, UK., Hinton JCD; Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, Liverpool, UK., Blower TR; Department of Biosciences, Durham University, South Road, Durham, UK. timothy.blower@durham.ac.uk.
Jazyk: angličtina
Zdroj: Nature communications [Nat Commun] 2024 Aug 22; Vol. 15 (1), pp. 7236. Date of Electronic Publication: 2024 Aug 22.
DOI: 10.1038/s41467-024-51629-7
Abstrakt: Bacteria have evolved a broad range of systems that provide defence against their viral predators, bacteriophages. Bacteriophage Exclusion (BREX) systems recognise and methylate 6 bp non-palindromic motifs within the host genome, and prevent replication of non-methylated phage DNA that encodes these same motifs. How BREX recognises cognate motifs has not been fully understood. In this study we characterise BREX from pathogenic Salmonella and present X-ray crystallographic structures of the conserved BREX protein, PglX. The PglX N-terminal domain encodes the methyltransferase, whereas the C-terminal domain is for motif recognition. We also present the structure of PglX bound to the phage-derived DNA mimic, Ocr, an inhibitor of BREX activity. Our analyses propose modes for DNA-binding by PglX and indicate that both methyltransferase activity and defence require larger BREX complexes. Through rational engineering of PglX we broaden both the range of phages targeted, and the host motif sequences that are methylated by BREX. Our data demonstrate that PglX is used to recognise specific DNA sequences for BREX activity, contributing to motif recognition for both phage defence and host methylation.
(© 2024. The Author(s).)
Databáze: MEDLINE
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