The integrase of genomic island GIsul2 mediates the mobilization of GIsul2 and ISCR-related element CR2-sul2 unit through site-specific recombination

In the worldwide health threat posed by antibiotic-resistant bacterial pathogens, mobile genetic elements (MGEs) play a critical role in favoring the dissemination of resistance genes. Among them, the genomic island GI sul2 and the IS CR -related element CR2 -sul2 unit are believed to participate in this dissemination. However, the mobility of the two elements has not yet been demonstrated. Here, we found that the GI sul2 and CR2 -sul2 units can excise from the host chromosomal attachment site ( attB ) in Shigella flexneri . Through establishing a two-plasmid mobilization system composed of a donor plasmid bearing the GI sul2 and a trap plasmid harboring the attB in recA -deficient Escherichia coli , we reveal that the integrase of GI sul2 can perform the excision and integration of GI sul2 and CR2 -sul2 unit by site-specific recombination between att core sites. Furthermore, we demonstrate that the integrase and the att sites are required for mobility through knockout experiments. Our findings provide the first experimental characterization of the mobility of GI sul2 and CR2 -sul2 units mediated by integrase. They also suggest a potential and unappreciated role of the GI sul2 integrase family in the dissemination of CR2 -sul2 units carrying various resistance determinants in between.