Structure and Rational Engineering of the PglX Methyltransferase and Specificity Factor for BREX Phage Defence

Overview

Journal Nat Commun

Specialty Biology

Date 2024 Aug 22

PMID 39174540

Authors

Sam C Went

David M Picton

Richard D Morgan

Andrew Nelson

Aisling Brady

Giuseppina Mariano

David T F Dryden

Darren L Smith

Nicolas Wenner

Jay C D Hinton

Tim R Blower

Affiliations

Soon will be listed here.

Abstract

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.

Citing Articles

Molecular basis of foreign DNA recognition by BREX anti-phage immunity system.

Drobiazko A, Adams M, Skutel M, Potekhina K, Kotovskaya O, Trofimova A Nat Commun. 2025; 16(1):1825.

PMID: 39979294 PMC: 11842806. DOI: 10.1038/s41467-025-57006-2.

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