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© Melanie Blokesch, EPFL
Flagellated Vibrio cholerae
Publication : Nucleic acids research

Replicative resolution of integron cassette insertion

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Nucleic acids research - 26 Jun 2012

Loot C, Ducos-Galand M, Escudero JA, Bouvier M, Mazel D

Link to Pubmed [PMID] – 22740653

Nucleic Acids Res. 2012 Sep;40(17):8361-70

Site-specific recombination catalyzed by tyrosine recombinases follows a common pathway consisting of two consecutive strand exchanges. The first strand exchange generates a Holliday junction (HJ), which is resolved by a second strand exchange. In integrons, attC sites recombine as folded single-stranded substrates. Only one of the two attC site strands, the bottom one, is efficiently bound and cleaved by the integrase during the insertion of gene cassettes at the double-stranded attI site. Due to the asymmetry of this complex, a second strand exchange on the attC bottom strand (bs) would form linearized abortive recombination products. We had proposed that HJ resolution would rely on an uncharacterized mechanism, probably replication. Using an attC site carried on a plasmid with each strand specifically tagged, we followed the destiny of each strand after recombination. We demonstrated that only one strand, the one carrying the attC bs, is exchanged. Furthermore, we show that the recombination products contain the attC site bs and its entire de novo synthesized complementary strand. Therefore, we demonstrate the replicative resolution of single-strand recombination in integrons and rule out the involvement of a second strand exchange of any kind in the attC×attI reaction.

http://www.ncbi.nlm.nih.gov/pubmed/22740653