title: Tn552 transposase catalyzes concerted strand transfer in vitro.
abstract: The Tn552 transposase, a member of the DDE superfamily of transposase and retroviral integrase proteins, has been expressed in soluble form. The purified protein performs concerted strand transfer in vitro, efficiently pairing two preprocessed transposon ends and inserting them into target DNA. For maximum efficiency, both participating DNA ends must contain the two adjacent transposase-binding sites that are the normal constituents of the Tn552 termini. As is the case with transposition in vivo, the insertions recovered from the reaction in vitro are flanked by repeats of a short target sequence, most frequently 6 bp. The reaction has stringent requirements for a divalent metal ion. Concerted strand transfer is most efficient with Mg2+. Although it stimulates strand transfer overall, Mn2+ promotes uncoupled, single-ended events at the expense of concerted insertions. The simplicity and efficiency of the Tn552 transposition system make it an attractive subject for structural and biochemical studies and a potentially useful genetic tool.
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520-8114, USA.
authors.forename: A E
authors.initials: AE
authors.lastname: Leschziner
authors.forename: T J
authors.initials: TJ
authors.lastname: Griffin
authors.forename: N D
authors.initials: ND
authors.lastname: Grindley
mesh_terms: D001483:Base Sequence; D002384:Catalysis; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism*; D004251:DNA Transposable Elements; D008274:Magnesium / Q000378:metabolism; D008345:Manganese / Q000378:metabolism; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000378:metabolism; D011489:Protein Denaturation; D012091:Repetitive Sequences, Nucleic Acid; D012995:Solubility; D019895:Transposases / Q000378:metabolism*
chemical_list: D004251:DNA Transposable Elements; D009841:Oligonucleotides; D008345:Manganese; D004247:DNA; D019895:Transposases; D008274:Magnesium