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| Subject Categories: Genome Stability & Dynamics |
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| The EMBO Journal
(2006) 25, 4586–4595, doi:10.1038/sj.emboj.7601325 Published online 14 September 2006 |
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Viewing single  site-specific recombination events from start to finish |
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| Jeffrey P Mumm1, 3, Arthur Landy1 and Jeff Gelles2 |
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1 Department of Molecular Biology, Cell Biology, and Biochemistry, Brown University, J Walter Wilson Laboratories, Providence, RI, USA 2 Department of Biochemistry, MS 009 Brandeis University, Waltham, MA, USA To whom correspondence should be addressed Arthur Landy, Department of Molecular Biology, Cell Biology, and Biochemistry, Brown University, J Walter Wilson Laboratories, room 360, 69 Brown Street, Providence, RI 02912, USA. Tel.: +1 401 863 2566; Fax: +1 401 863 1348; E-mail: arthur_landy@brown.edu 3 Present address: Department of Biochemistry, MS 009 Brandeis University, Waltham, MA 02454-9110, USA Received 22 May 2006; Accepted 10 August 2006; Published online 14 September 2006. |
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| Abstract |
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The site-specific recombination pathway by which the bacteriophage chromosome is excised from its Escherichia coli host chromosome is a tightly regulated, highly directional, multistep reaction that is executed by a series of multiprotein complexes. Until now, it has been difficult to study the individual steps of such reactions in the context of the entire pathway. Using single-molecule light microscopy, we have examined this process from start to finish. Stable bent-DNA complexes containing integrase and the accessory proteins IHF (integration host factor) and Xis form rapidly on attL and attR recombination partners, and synapsis of partner complexes follows rapidly after their formation. Integrase-mediated DNA cleavage before or immediately after synapsis is required to stabilize the synaptic assemblies. Those complexes that synapsed ( 50% of the total) yield recombinant product with a remarkable 100% efficiency. The rate-limiting step of excision occurs after synapsis, but closely precedes or is concomitant with the appearance of a stable Holliday junction. Our kinetic analysis shows that directionality of this recombination reaction is conferred by the irreversibility of multiple reaction steps. |
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| Keywords: bacteriophage , DNA recombination, Holliday junction, synapsis, tyrosine recombinase |
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Top of page MORE ARTICLES LIKE THISThese links to content published by NPG are automatically generated NEWS AND VIEWSNature News and Views (24 Sep 1987) DNA recombination: Resolution of intermediates Nature News and Views (25 Oct 1984) RESEARCHViewing single λ site-specific recombination events from start to finish The EMBO Journal Article A structural basis for allosteric control of DNA recombination by λ integrase Nature Article (23 Jun 2005) |
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