Transposon Tn3 encodes a site-specific recombination system: identification of essential sequences, genes, and actual site of recombination.

Kostriken, Richard; Morita, Chika; Heffron, Fred

doi:10.1073/pnas.78.7.4041

Cited by 63 publications

(25 citation statements)

References 21 publications

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“…As in other characterized sitespecific recombination systems, the recombination enzyme, resolvase, is encoded by a gene (tnpR) adjacent to the recombination site (res). Tn3 resolvase and res are both similar to and interchangeable with the site-specific recombination ' determinants specified by the related transposable element -y6 (Kostriken et al, 1981;Reed, 1981a;Grindley et al, 1982;Kitts et al, 1982b). Both resolvases act rapidly and efficiently on directly repeated res regions in the same molecule but appear to act poorly in the reverse fusion reaction and on inverted substrates in the same molecule (Chiang and Clowes, 1982;Reed and Grindley, 1981).…”

Section: Introductionmentioning

confidence: 99%

Transposon-encoded site-specific recombination: nature of the Tn3 DNA sequences which constitute the recombination site res.

Kitts¹,

Symington²,

Dyson³

et al. 1983

The EMBO Journal

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The tnpR gene of transposon Tn3 encodes a site-specific recombination enzyme that acts at res, a DNA region adjacent to tnpR, to convert co-integrate intermediates of interreplicon transposition to the normal transposition end-products. We have used two complementary approaches to study the nature of the Tn3 recombination region, res. Firstly, the DNA-binding sites for tnpR protein were determined in DNase I protection experiments. These identified a 120-bp region between the tnpA and tnpR genes that can be subdivided into three separate protein-binding sites. Genetic dissection experiments indicate that few, if any, other sequences in addition to this 120-bp region are required for res function. Moreover, we have shown that the two directly repeated res regions within a molecule are unequal partners in the recombination reaction: a truncated res region, which is unable to recombine with a second identical res region, can recombine efficiently with an intact res region. This demonstration, along with the observation that tnpR/res recombination acts efficiently on directly repeated res regions within a molecule but inefficiently both on inverted res regions in the same molecule and in the fusion reaction between res regions in different molecules, leads us to propose that onedimensional diffusion (tracking) of tnpR protein along DNA is used to locate an initial res region, and then to bring a second directly repeated res region into a position that allows recombination between the res regions.

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Section: Introductionmentioning

confidence: 99%

Transposon-encoded site-specific recombination: nature of the Tn3 DNA sequences which constitute the recombination site res.

Kitts¹,

Symington²,

Dyson³

et al. 1983

The EMBO Journal

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“…The tnpA gene encodes transposase, which recognizes the terminal inverted repeat sequences (IRR and IRL) and catalyses the first transposition step, which is the formation of a cointegrate between a donor replicon carrying Tn3 and a target replicon (Gill et al 1979;Heffron et al 1979;McCormick et al 1981;Ichikawa et al 1987;). The tnpR gene encodes resolvase, which resolves the cointegrate molecule efficiently into two products, both carrying Tn3 , by acting at the res site (Kostriken et al, 1981;Reed 1981a,b). Tn3 transposes to a target replicon with Tn3, less frequently than to a target replicon with no Tn3, and this phenomenon has been called transposition immunity (Robinson et al 1977).…”

Section: Introductionmentioning

confidence: 99%

A cell‐free system of Tn3 transposition and transposition immunity

1996

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“…TnpA protein binds nonspecific DNA sequences with a preference for single-stranded DNA (19), specifically binds fragments containing an end of Tn3 (31,47), and forms multimeric complexes in vitro (19). In the second stage, the cointegrate structure is separated (resolved) by a TnpRmediated site-specific recombination between the duplicated copies of Tn3 (3,32,35). The resolution site (res) is required in cis for TnpR-mediated regulation and is the site of both transcriptional repression and recombination (22,27,35,49,50).…”

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confidence: 99%

Nucleotide sequences required for Tn3 transposition immunity

Kans

Casadaban

1989

J Bacteriol

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The Tn3 transposon inserts at a reduced frequency into a plasmid already containing a copy of Tn3, a phenomenon known as transposition immunity. The cis-acting site on Tn3 responsible for immunity was mapped by deletions from each side to be within the terminal 38-base-pair sequence that is inversely repeated at the ends of Tn3. Two palindromic sequences are present in the essential part of this region. Some deletions conferred only partial immunity, and others conferred negative immunity. Multiple copies of partially immune ends conferred additional immunity. No other part of Tn3 was necessary for immunity.

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Transposon Tn3 encodes a site-specific recombination system: identification of essential sequences, genes, and actual site of recombination.

Abstract: The bacterial transposon Tn3 encodes a site-specific recombination system. The recombination requires the product oftnpR, a gene previously identified as a repressor ofthe transposase. This recombination is site specific and takes place somewhere within the sequence

Cited by 63 publications

References 21 publications

Transposon-encoded site-specific recombination: nature of the Tn3 DNA sequences which constitute the recombination site res.

Transposon-encoded site-specific recombination: nature of the Tn3 DNA sequences which constitute the recombination site res.

A cell‐free system of Tn3 transposition and transposition immunity

Nucleotide sequences required for Tn3 transposition immunity

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