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• W2048103135 abstract "Background: A fundamental feature of bacteriophage λ site-specific recombination is the strict requirement for a region of sequence identity between recombining DNA duplexes. It has been difficult to understand how the recombination machinery identifies and responds to non-homologies as subtle as a single base-pair substitution, because the reaction intermediates are transient and there are likely to be several different homology-dependent steps. In order to understand better how the recombination machinery compares parental sequences, we have used the recently developed ‘suicide substrates’ — DNA containing 5′-bridging phosphorothioate linkages — to monitor the timing of homology-sensing relative to the strand cleavage reactions.Results The cleavage reactions for the two different strands of attB, the bacterial recombination locus for λ integration, show very different degrees of dependence on homology with the partner locus, attP. Strand cleavage at the B binding site for Int recombinase is insensitive to homology. In contrast, cleavage at the B′ binding site strongly depends on homology in the three base pairs adjacent to the B site. Strand cleavage at the B site is apparently required for the readout of this homology but, surprisingly, joining of the cleaved B site to a partner is not.Conclusion Our finding that cleavage at the B site is insensitive to homology shows that effective synapsis between partners does not depend on sequence matching. Cleavage at the B′ site provides the earliest positive signal for a homology-dependent switch in the λ recombination machinery. Because this switch can occur in the absence of strand joining, the results argue against models that invoke strand ligation as the critical element of homology-sensing. Alternative mechanisms are presented that involve varieties of non-covalent strand swapping. A synthesis of the present results and other recent experiments highlights the importance of the disannealing of complementary strands and their reannealing to new partners, a process traditionally described as branch migration. The reversibility of branch migration and its bias away from mismatched combinations are proposed to be the major mechanisms of homology-sensing during λ integration." @default.
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• W2048103135 date "1995-11-01" @default.
• W2048103135 modified "2023-09-30" @default.
• W2048103135 title "Suicide substrates reveal properties of the homology-dependent steps during integrative recombination of bacteriophage λ" @default.
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• W2048103135 doi "https://doi.org/10.1016/s0960-9822(95)00258-2" @default.
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