A minimal system for Tn7 transposition: the transposon-encoded proteins TnsA and TnsB can execute DNA breakage and joining reactions that generate circularized Tn7 species.
In the presence of ATP and Mg(2+), the bacterial transposon Tn7 translocates via a cut and paste mechanism executed by the transposon-encoded proteins TnsA+TnsB+TnsC+TnsD. We report here that in the presence of Mn(2+), TnsA+TnsB alone can execute the DNA breakage and joining reactions of Tn7 recombination. ATP is not essential in this minimal system, revealing that this cofactor is not directly involved in the chemical steps of recombination. In both the TnsAB and TnsABC+D systems, recombination initiates with double-strand breaks at each transposon end that cut Tn7 away from flanking donor DNA. In the minimal system, breakage occurs predominantly at a single transposon end and the subsequent end-joining reactions are intramolecular, with the exposed 3' termini of a broken transposon end joining near the other end of the Tn7 element in the same donor molecule to form circular transposon species. In contrast, in TnsABC+D recombination, breaks occur at both ends of Tn7 and the two ends join to a target site on a different DNA molecule to form an intermolecular simple insertion. This demonstration of the capacity of TnsAB to execute breakage and joining reactions supports the view that these proteins form the Tn7 transposase.
Biery
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MC
Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Lopata
M
M
Craig
N L
NL
D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D002413:Cations, Divalent / Q000494:pharmacology; D015342:DNA Probes; D004251:DNA Transposable Elements / Q000235:genetics* / Q000502:physiology; D004270:DNA, Circular / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism / Q000648:ultrastructure; D004278:DNA, Superhelical / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000648:ultrastructure; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D029968:Escherichia coli Proteins; D008345:Manganese / Q000494:pharmacology; D008854:Microscopy, Electron; D008970:Molecular Weight; D009154:Mutation / Q000187:drug effects / Q000235:genetics; D009690:Nucleic Acid Conformation; D009711:Nucleotides / Q000235:genetics; D011995:Recombination, Genetic / Q000187:drug effects / Q000235:genetics*
D001426:Bacterial Proteins; D002413:Cations, Divalent; D015342:DNA Probes; D004251:DNA Transposable Elements; D004270:DNA, Circular; D004278:DNA, Superhelical; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D009711:Nucleotides; C098908:TnsA protein, E coli; C069633:TnsB protein, E coli; D008345:Manganese