The rulB gene of plasmid pWW0 is a hotspot for the site-specific insertion of integron-like elements found in the chromosomes of environmental Pseudomonas fluorescens group bacteria

The rulAB operon of Pseudomonas spp. confers fitness traits on the host and has been suggested to be a hotspot for insertion of mobile elements that carry avirulence genes. Here, for the first time, we show that rulB on plasmid pWW0 is a hotspot for the active site-specific integration of related integron-like elements (ILEs) found in 6 environmental pseudomonads (strains FH1-6). Integration into rulB on pWW0 occurred at position 6488 generating a 3 bp direct repeat. ILEs from FH1 and FH5 were 9403 in length and contained 8 ORFs whilst the ILE from FH4 was 16233 bp in length and contained 16 ORFs. In all three ILEs the first 5.1 kb (containing ORFs 1-4) were structurally conserved and contained 3 predicted site-specific recombinases/integrases and a tetR homologue.Downstream of these resided ORFs of the ‘variable side’ with structural and sequence similarity to those encoding survival traits on the fitness enhancing plasmid pGRT1 (ILEFH1 and ILEFH5) and the NR-II virulence region of genomic island PAGI-5 (ILEFH4). Collectively, these ILEs share features with the previously described type III protein secretion system effector (T3SE) integron-like elements and are considered important to host survival and transfer of fitness enhancing and (a)virulence genes between bacteria.