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Single-molecule analysis reveals the molecular bearing mechanism of DNA strand exchange by a serine recombinase

Bai, H and Sun, M and Ghosh, P and Hatfull, GF and Grindley, NDF and Marko, JF (2011) Single-molecule analysis reveals the molecular bearing mechanism of DNA strand exchange by a serine recombinase. Proceedings of the National Academy of Sciences of the United States of America, 108 (18). 7419 - 7424. ISSN 0027-8424

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Abstract

Structural and topological data suggest that serine site-specific DNA recombinases exchange duplex DNAs by rigid-body relative rotation of the two halves of the synapse, mediated by a flat protein-protein interaction surface. We present evidence for this rotational motion for a simple serine recombinase, the Bxb1 phage integrase, from a single-DNA-based supercoil-release assay that allows us to follow crossover site cleavage, rotation, religation, and product release in real time. We have also used a two-DNA braiding-relaxation experiment to observe the effect of synapse rotation in reactions on two long molecules. Relaxation and unbraiding are rapid (averaging 54 and 70 turns/s, respectively) and complete, with no discernible pauses. Nevertheless, the molecular friction associated with rotation is larger than that of type-I topoisomerases in a similar assay. Surprisingly we find that the synapse can stay rotationally "open" for many minutes.


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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Bai, H
Sun, M
Ghosh, P
Hatfull, GFgfh@pitt.eduGFH
Grindley, NDF
Marko, JF
Date: 3 May 2011
Date Type: Publication
Journal or Publication Title: Proceedings of the National Academy of Sciences of the United States of America
Volume: 108
Number: 18
Page Range: 7419 - 7424
DOI or Unique Handle: 10.1073/pnas.1018436108
Schools and Programs: Dietrich School of Arts and Sciences > Biological Sciences
Refereed: Yes
ISSN: 0027-8424
MeSH Headings: Biotin; DNA Nucleotidyltransferases--metabolism; DNA, Superhelical--chemistry; DNA, Superhelical--metabolism; Digoxigenin; Escherichia coli--genetics; Plasmids--genetics; Rotation; Serine--metabolism
Other ID: NLM PMC3088605
PubMed Central ID: PMC3088605
PubMed ID: 21502527
Date Deposited: 29 Oct 2012 20:29
Last Modified: 02 Feb 2019 15:57
URI: http://d-scholarship.pitt.edu/id/eprint/16115

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