IS-Linked Movement of a Restriction-Modification SystemReport as inadecuate

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Potential mobility of restriction-modification systems has been suggested by evolutionary-bioinformatic analysis of prokaryotic genomes. Here we demonstrate in vivo movement of a restriction-modification system within a genome under a laboratory condition. After blocking replication of a temperature-sensitive plasmid carrying a PaeR7I restriction-modification system in Escherichia coli cells, the plasmid was found integrated into the chromosome of the surviving cells. Sequence analysis revealed that, in the majority of products, the restriction-modification system was linked to chromosomal insertion sequences ISs. Three types of products were: I apparent co-integration of the plasmid and the chromosome at a chromosomal IS1 or IS5 copy 24-28 analyzed; II de novo insertion of IS1 with the entire plasmid except for a 1–3 bp terminal deletion 2-28; and III reciprocal crossing-over between the plasmid and the chromosome involving 1–3 bp of sequence identity 2-28. An R-negative mutation apparently decreased the efficiency of successful integration by two orders of magnitude. Reconstruction experiments demonstrated that the restriction-dependence was mainly due to selection against cells without proper integration: their growth was inhibited by the restriction enzyme action. These results demonstrate collaboration of a mobile element and a restriction-modification system for successful joint migration. This collaboration may have promoted the spread and, therefore, the long-term persistence of these complexes and restriction-modification systems in a wide range of prokaryotes.

Author: Noriko Takahashi, Seishi Ohashi, Marat R. Sadykov, Yoko Mizutani-Ui, Ichizo Kobayashi



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