Abstract
Genomic islands are DNA regions containing variable genetic information related to secondary metabolism. Frequently, they have the ability to excise from and integrate into replicons through site-specific recombination. Thus, they are usually flanked by short direct repeats that act as attachment sites, and contain genes for an integrase and an excisionase which carry out the genetic exchange. These mobility events would be at the basis of the horizontal transfer of genomic islands among bacteria.Microcin H47 is a ribosomally-synthesized antibacterial peptide that belongs to the group of chromosome-encoded microcins. The 13 kb-genetic system responsible for its production resides in the chromosome of the Escherichia coli H47 strain and is flanked by extensive and imperfect direct repeats. In this work, both excision and integration of the microcin H47 system were experimentally detected. The analyses were mainly performed in E. coli K12 cells carrying the microcin system cloned in a multicopy plasmid. As expected of a site-specific recombination event, the genetic exchange also occurred in a context deficient for homologous recombination. The DNA sequence of the attachment sites resulting from excision were hybrid between the sequences of the direct repeats. Unexpectedly, different hybrid attachment sites appeared which resulted from recombination in four segments of identity between the direct repeats. Genes encoding the trans-acting proteins responsible for the site-specific recombination were shown to be absent in the microcin H47 system. Therefore, they should be provided by the remaining genetic context, not only in the H47 strain but also in E. coli K12 cells, where both excision and integration occurred. Moreover, a survey of the attachment sites in data banks revealed that they are widely spread among E. coli strains. It is concluded that the microcin system is a small island –H47 genomic island- that would employ a parasitic strategy for its mobility.
Highlights
The increasing sequencing of complete bacterial genomes revealed the existence of a flexible genetic content that varies even among closely related strains
Most genomic islands (GI) are flanked by short direct repeats that act as attachment sites left and right and contain the genes for the corresponding integrase and excisionase [4,5,6,7]
It could be present in the E. coli strain CA46, which contains the microcin H47 (MccH47) system limited by an attL-like sequence
Summary
The increasing sequencing of complete bacterial genomes revealed the existence of a flexible genetic content that varies even among closely related strains In the chromosome, this variable genetic information is gathered in defined regions of up to 200 kb called genomic islands (GI). The mobility of GIs reproduces the site-specific recombination events characteristic of temperate phages following the lisogenic pathway. It requires the presence of attachment sites and the participation of specific recombination functions carried out by an integrase and an excisionase. The island integrated into the chromosome excises through site-specific recombination between its flanking repeats, a process operated by the corresponding integrase-excisionase pair. The original condition of the GI is reproduced, but in the context of a new host
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