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Control of directionality in L5 integrase-mediated site-specific recombination

Lewis, JA and Hatfull, GF (2003) Control of directionality in L5 integrase-mediated site-specific recombination. Journal of Molecular Biology, 326 (3). 805 - 821. ISSN 0022-2836

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Abstract

Mycobacteriophage L5 is a temperate phage that forms lysogens in Mycobacterium smegmatis. These lysogens carry an integrated L5 prophage inserted at a specific chromosomal location and undergo subsequent excision during induction of lytic growth. Both the integrative and excisive site-specific recombination events are catalyzed by the phage-encoded tyrosine integrase (Int-L5) and require the host-encoded protein, mIHF. The directionality of these recombination events is determined by a second phage-encoded protein, Excise, the product of gene 36 (Xis-L5); integration occurs efficiently in the absence of Xis-L5 while excision is dependent upon it. We show here that Xis-L5 binds to attR DNA, introduces a DNA bend, and facilitates the formation of an intasome-R complex. This complex, which requires mIHF, Xis-L5 and Int-L5, readily recombines with a second intasome formed by Int-L5, mIHF and attL DNA (intasome-L) to generate the attP and attB products of excision. Xis-L5 also strongly inhibits Int-L5-mediated integrative recombination but does not prevent either the protein-DNA interactions that form the attP intasome (intasome-P) or the capture of attB, but acts later in the reaction presumably by preventing the formation of a recombinagenic synaptic intermediate. The mechanism of action of Xis-L5 appears to be purely architectural, influencing the assembly of protein-DNA structures solely through its DNA-binding and DNA-bending properties. © 2003 Elsevier Science Ltd. All rights reserved.

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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Lewis, JA
Hatfull, GFgfh@pitt.eduGFH
Date: 21 February 2003
Date Type: Publication
Journal or Publication Title: Journal of Molecular Biology
Volume: 326
Number: 3
Page Range: 805 - 821
DOI or Unique Handle: 10.1016/s0022-2836(02)01475-4
Schools and Programs: Dietrich School of Arts and Sciences > Biological Sciences
Refereed: Yes
ISSN: 0022-2836
MeSH Headings: Bacterial Proteins; Base Sequence; DNA--metabolism; DNA Footprinting; Integrases--physiology; Membrane Proteins--genetics; Molecular Sequence Data; Plasmids; Recombination, Genetic--physiology
PubMed ID: 12581642
Date Deposited: 19 Nov 2012 16:57
Last Modified: 02 Feb 2019 15:57
URI: http://d-scholarship.pitt.edu/id/eprint/16354

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