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Synapsis in phage Bxb1 integration: Selection mechanism for the correct pair of recombination sites

Ghosh, P and Pannunzio, NR and Hatfull, GF and Gottesman, M (2005) Synapsis in phage Bxb1 integration: Selection mechanism for the correct pair of recombination sites. Journal of Molecular Biology, 349 (2). 331 - 348. ISSN 0022-2836

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Abstract

Recombination by site-specific recombinases is a highly concerted process that requires synapsis of the correct pair of DNA substrates. Phage-encoded serine-integrases are unusual among the serine-recombinase family, which includes transposon resolvases and DNA invertases, in that they utilize two simple but different DNA substrates (attB and attP) and do not require accessory sites, additional proteins, or DNA supercoiling. Synapsis must therefore be directed solely by integrase-DNA interactions. We show here that the Bxb1 serine-integrase binds as a dimer to its two DNA substrates (attB, attP) and recombinant products (attL, attR) with similar affinities. However, synapsis occurs only between attP and attB, and not between any of the other nine possible site combinations. The Bxb1 integrase domain structure, the unusual DNA-binding properties of the integrase, and the characterization of a mutant protein with altered site-discrimination, are consistent with synaptic selectivity being derived from DNA sequence-induced changes in the conformations of integrase-DNA complexes. © 2005 Elsevier Ltd. All rights reserved.


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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Ghosh, P
Pannunzio, NR
Hatfull, GFgfh@pitt.eduGFH
Gottesman, M
Date: 3 June 2005
Date Type: Publication
Journal or Publication Title: Journal of Molecular Biology
Volume: 349
Number: 2
Page Range: 331 - 348
DOI or Unique Handle: 10.1016/j.jmb.2005.03.043
Schools and Programs: Dietrich School of Arts and Sciences > Biological Sciences
Refereed: Yes
ISSN: 0022-2836
MeSH Headings: Amino Acid Sequence; Attachment Sites, Microbiological--genetics; Base Pairing--genetics; Base Sequence; DNA--genetics; DNA--metabolism; Dimerization; Integrases--metabolism; Molecular Sequence Data; Molecular Weight; Mutation--genetics; Mycobacteriophages--enzymology; Mycobacteriophages--genetics; Protein Binding; Recombinases--chemistry; Recombinases--metabolism; Recombination, Genetic--genetics; Sequence Alignment; Substrate Specificity; Tyrosine--genetics; Tyrosine--metabolism; Virus Integration--genetics
PubMed ID: 15890199
Date Deposited: 15 Nov 2012 20:22
Last Modified: 02 Feb 2019 15:57
URI: http://d-scholarship.pitt.edu/id/eprint/16224

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