The bacteroides NBU1 integrase performs a homology-independent strand exchange to form a holliday junction intermediate.

The bacteroides NBU1 integrase performs a homology-independent strand exchange to form a holliday junction intermediate. Research Abstract Details 

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The bacteroides NBU1 integrase performs a homology-independent strand exchange to form a holliday junction intermediate. Abstract Text:

Lara Rajeev,Anca Segall,Jeffrey Gardner,Lara Rajeev,Anca Segall,Jeffrey Gardner,

The Bacteroides mobilizable transposon NBU1 uses an integrase (IntN1) that is a tyrosine recombinase for its integration and excision from the host chromosome. Previously we showed that IntN1 makes 7-bp staggered cuts within the NBU1 att sites, and certain mismatches within the crossover region of the attN1 site (G(-2)C attN1) or the chromosomal target site (C(-3)G attBT1-1) enhanced the in vivo integration efficiency. Here we describe an in vitro integration system for NBU1. We used nicked substrates and a Holliday junction trapping peptide to show that NBU1 integration proceeds via formation of a Holliday junction intermediate that is formed by exchange of bottom strands. Some mismatches next to the first strand exchange site (in reactions with C(-3)G attBT1-1 or G(-2)C attN1 with their wild-type partner site) not only allowed formation of the Holliday junction intermediate but also increased the rate of recombinant formation. The second strand exchange appears to be homology-dependent. IntN1 is the only tyrosine recombinase known to catalyze a reaction that is more efficient in the presence of mismatches and where the first strand exchange is homology-independent. The possible mechanisms by which the mismatches stimulate recombination are discussed.

The bacteroides NBU1 integrase performs a homology-independent strand exchange to form a holliday junction intermediate. Publishing Authors By Initials

L Rajeev,A Segall,J Gardner,L Rajeev,A Segall,J Gardner,

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The bacteroides NBU1 integrase performs a homology-independent strand exchange to form a holliday junction intermediate. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: The Journal of biological chemistry

VOLUME: 282

Page Numbers: 31228-37

Journal Abbreviation: J. Biol. Chem.

ISSN: 0021-9258

DAY: 30

MONTH: 08

YEAR: 2007

The bacteroides NBU1 integrase performs a homology-independent strand exchange to form a holliday junction intermediate. Information

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LANGUAGE: eng

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Grant and Affiliation Information for The bacteroides NBU1 integrase performs a homology-independent strand exchange to form a holliday junction intermediate.

AFFILIATION: Department of Microbiology, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA. rajeev@uiuc.edu

Country: United States

AGENCY: United States NIGMS

GRANT: GM52847

ACRONYM: GM

MEDLINETA: J Biol Chem

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