Tn10 and IS10 Transposition and Chromosome Rearrangements: Mechanism and Regulation In Vivo and In Vitro

Kleckner, Nancy; Chalmers, Ronald; Kwon, Douglas S.; Sakai, J.; Bolland, Silvia

doi:10.1007/978-3-642-79795-8_3

Cited by 85 publications

(103 citation statements)

References 92 publications

Supporting

Mentioning

101

Contrasting

Order By: Relevance

“…1D, Kaufman & Rio 1992;van Luenen et al 1994;Craig 1996;Kleckner et al 1996;Vos et al 1996). Double strand cleavage at both ends is obligatory prior to target DNA capture and strand transfer for Tn10 .…”

Section: Classical Transposition Reactionsmentioning

confidence: 99%

Polynucleotidyl transfer reactions in site‐specific DNA recombination

1997

View full text Add to dashboard Cite

Site-specific DNA rearrangement reactions are widespread among organisms. They are used, for example, by vertebrates to boost immune response diversity, and in turn by parasitic organisms to evade the host immune system by surface antigen switching. Parasitic genetic elements ubiquitous to most organisms invade new host genomic sites by a variety of types of site-specific recombination. Polynucleotidyl transfer reactions are central to these DNA recombination reactions. The recombinase of each reaction system that 'catalyses' such chemical reactions at specific DNA sites are apparently designed to accomplish unique DNA geometrical specificity, or delicate control over the extent or direction of the reaction, with the sacrifice of protein turnover. Here we discuss our current understanding of several issues that relate to the polynucleotidyl transfer steps in several of the better studied site-specific recombination reactions.

show abstract

Section: Classical Transposition Reactionsmentioning

confidence: 99%

Polynucleotidyl transfer reactions in site‐specific DNA recombination

1997

View full text Add to dashboard Cite

show abstract

“…The excised transposon is then inserted (by singlestrand connections) into target DNA. Both Tn7 and Tn10 use this mechanism (see Craig, 1996;Kleckner et al, 1996). Transposons that make only single-strand cuts and joins generally make co-integrates (e.g.…”

Section: A Model For Is 2 Intermolecular Transpositionmentioning

confidence: 99%

“…Recent studies of specific members of the IS3 family suggest that their transposition may differ significantly from the two classical paradigms established for Mu, Tn10 and Tn7 (Mizuuchi, 1992;Craig, 1996;Kleckner et al, 1996). In vitro studies of these three elements have shown that transposases either cut both DNA strands at the transposon's two ends to excise it completely from its vector (e.g.…”

Section: Introductionmentioning

confidence: 99%

Two abundant intramolecular transposition products, resulting from reactions initiated at a single end, suggest that IS2 transposes by an unconventional pathway

Lewis

Grindley

1997

Molecular Microbiology

View full text Add to dashboard Cite

SummaryThe Escherichia coli insertion sequence, IS2, is a member of the IS3 family of bacterial transposable elements. Its transposase is a fusion protein, OrfAB, made by a programmed ¹1 translational frameshift near to the end of orfA and just after the start of orfB. We have characterized two major products of IS2 intramolecular transposition, which accumulate in cells that express the IS2 OrfAB fusion protein at elevated levels. The more abundant product is a minicircle composed of the complete IS2 with just a single basepair (occasionally 2 bp) separating the two IS ends. In all cases, this basepair is derived from the vector sequence immediately adjacent to the left IS2 end (IRL). The second product is a figure-eight molecule that contains all the IS2 and vector sequences present in the parental plasmid. One DNA strand contains the parental sequences unrearranged. The other contains a single-stranded version of the minicircle junction -the precise 3Ј end of IRR has been cleaved and joined to a target just outside the 5Ј end of IRL; the remaining vector sequences have a free 5Ј end, derived from cleavage at the 3Ј end of IRR, and a free 3Ј end, released upon cleavage of the target site adjacent to IRL. We propose that figure-eight molecules are the precursor to IS2 minicircles and that the formation of these two products is the initial step in IS2 intermolecular transposition. This proposed transposition pathway provides a means for a transposase that can cleave only one strand at each IS end to produce simple insertions and avoid forming co-integrates.

show abstract

“…IS10 ; see Kleckner et al, 1996). Since bacteria occupy a multitude of diverse ecological niches and receive signals and environmental stresses from a wide range of sources, it would not be surprising if certain of these signals exercised an influence on transposition activity.…”

Section: Introductionmentioning

confidence: 99%

IS911‐mediated intramolecular transposition is naturally temperature sensitive

Haren

Bétermier

Polard

et al. 1997

Molecular Microbiology

View full text Add to dashboard Cite

SummaryIt is shown here that the bacterial insertion sequence IS911 exhibits a temperature-sensitive transposition phenotype. Previous results have demonstrated that elevated levels of the IS911 transposase OrfAB generate significant quantities of a figure-eight form, created by cleavage and circularization of one of the transposon strands, and of an excised circular form, in which both transposon strands have been circularized. We show here that the level of both types of molecule observed in vivo was greatly reduced at 42ЊC compared with 37ЊC. On the other hand, reducing the temperature to 30ЊC resulted in a significant increase in production. Transposition activity at this temperature was sufficiently high to permit detection in vivo of an excised circular form of a defective single IS911 chromosomal copy when OrfAB is supplied in trans. A similar temperature-activity profile is observed for a cell-free reaction that uses partially purified OrfAB and generates the figure-eight form uniquely. Moreover, two point mutants of OrfAB were obtained, which render the reactions partially temperature resistant both in vivo and in vitro. These results suggest that some property of transposase itself is sensitive to elevated temperatures.

show abstract

Tn10 and IS10 Transposition and Chromosome Rearrangements: Mechanism and Regulation In Vivo and In Vitro

Cited by 85 publications

References 92 publications

Polynucleotidyl transfer reactions in site‐specific DNA recombination

Polynucleotidyl transfer reactions in site‐specific DNA recombination

Two abundant intramolecular transposition products, resulting from reactions initiated at a single end, suggest that IS2 transposes by an unconventional pathway

IS911‐mediated intramolecular transposition is naturally temperature sensitive

Contact Info

Product

Resources

About