Type I BREX system defends against antibiotic-resistant plasmids in Escherichia coli .

The Bacteriophage Exclusion (BREX) system is a novel antiphage defense system identified in Bacillus cereus in 2015. The purpose of this study was to investigate the presence of the BREX system defenses against antibiotic-resistant plasmids such as bla KPC and bla NDM invasion in Escherichia coli . The BREX system was present in 5.4% (23/424) of E. coli clinical isolates and 6.5% (84/1283) of E. coli strains with completely sequenced genomes in the GenBank database. All 23 BREX-positive E. coli clinical isolates were susceptible to carbapenems, while all five isolates carrying bla KPC and 11 carrying bla NDM were BREX-negative. For E. coli strains in the GenBank database, 37 of 38 strains carrying bla KPC and 109 of 111 strains carrying bla NDM were BREX negative. The recognition site sequence of methyltransferase PglX in a clinical E. coli 3756 was 5'-CANC A TC-3' using PacBio single-molecular real-time sequencing. The transformation efficiency of plasmid psgRNA-ColAori-target with the PglX recognition site was reduced by 100% compared with the plasmid without the recognition site in E. coli DH5α-pHSG398-BREX. The BREX showed lower defense efficacy against plasmid psgRNA-15Aori-target which had the same plasmid backbone but different surrounding sequences of recognition sites with psgRNA-ColAori-target. The conjugation frequency of the KPC-2 plasmid and NDM-5 plasmid in E. coli 3756-ΔBREX was higher than that in E. coli 3756 clinical isolate (1.0 × 10 -6 vs 1.3 × 10 -7 and 5.5 × 10 -7 vs 1.7 × 10 -8 , respectively). This study demonstrated that the type I BREX system defends against antibiotic-resistant plasmids in E. coli .