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Resolvase-catalysed reactions between res sites differing in the central dinucleotide of subsite I

Stark, WM and Grindley, NDF and Hatfull, GF and Boocock, MR (1991) Resolvase-catalysed reactions between res sites differing in the central dinucleotide of subsite I. EMBO Journal, 10 (11). 3541 - 3548. ISSN 0261-4189

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The resolvase-catalysed reaction between two res sites in a circular DNA substrate normally gives two circular recombination products linked in a two-noded catenane. Homology between the two res sites at the central overlap dinucleotide of subsite I is important for recombination. Reactions between res sites differing at one position in the central dinucleotide (AC x AT) gave a low yield of recombinants containing mismatched base-pairs, but gave large amounts of a non-recombinant four-noded knot. This result was predicted by a 'simple rotation' model for strand exchange. The mismatch is evidently recognized only after commitment to an initial 180° rotation of the resolvase-linked DNA ends, and it induces a second 180° rotation which restores correct base-pairing at the overlap, giving the four-noded product. Correct base-pairing is not essential for religation, but may be important for release of the products. Characteristic patterns of 4, 6, 8 and 10 node knots, or 4, 8, 12 and 16 node knots were obtained, depending on the reaction conditions and the resolvase. Two pathways for multiple rounds of rotation in 360° steps are inferred. The results support a model for strand exchange by supercoil-directed subunit rotation within a resolvase tetramer.


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Item Type: Article
Status: Published
CreatorsEmailPitt UsernameORCID
Stark, WM
Grindley, NDF
Hatfull, GFgfh@pitt.eduGFH
Boocock, MR
Date: 1 January 1991
Date Type: Publication
Journal or Publication Title: EMBO Journal
Volume: 10
Number: 11
Page Range: 3541 - 3548
DOI or Unique Handle: 10.1002/j.1460-2075.1991.tb04918.x
Schools and Programs: Dietrich School of Arts and Sciences > Biological Sciences
Refereed: Yes
ISSN: 0261-4189
MeSH Headings: Base Sequence; Catalysis; DNA--genetics; DNA--metabolism; Electrophoresis, Agar Gel; Glycoside Hydrolases--metabolism; Kinetics; Molecular Sequence Data; Nucleic Acid Conformation; Nucleotidyltransferases--metabolism; Plasmids; Recombination, Genetic; Substrate Specificity; Transposases; beta-Fructofuranosidase
Other ID: NLM PMC453083
PubMed Central ID: PMC453083
PubMed ID: 1655422
Date Deposited: 12 Mar 2013 19:33
Last Modified: 02 Aug 2020 13:58


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