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
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Status: |
Published |
Creators/Authors: |
Creators | Email | Pitt Username | ORCID  |
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Bai, H | | | | Sun, M | | | | Ghosh, P | | | | Hatfull, GF | gfh@pitt.edu | GFH | | Grindley, NDF | | | | Marko, JF | | | |
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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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