Promiscuous target interactions in the mariner transposon Himar1.
We have previously characterized the early intermediates of mariner transposition. Here we characterize the target interactions that occur later in the reaction. We find that, in contrast to the early transposition intermediates, the strand transfer complex is extremely stable and difficult to disassemble. Transposase is tightly bound to the transposon ends constraining rotation of the DNA at the single strand gaps in the target site flanking the element on either side. We also find that although the cleavage step requires Mg2+ or Mn2+ as cofactor, the strand transfer step is also supported by Ca2+, suggesting that the structure of the active site changes between cleavage and insertion. Finally, we show that, in contrast to the bacterial cut and paste transposons, mariner target interactions are promiscuous and can take place either before or after cleavage of the flanking DNA. This is similar to the behavior of the V(D)J system, which is believed to be derived from an ancestral eukaryotic transposon. We discuss the implications of promiscuous target interactions for promoting local transposition and whether this is an adaptation to facilitate the invasion of a genome following horizontal transfer to a new host species.
Lipkow
Karen
K
Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom.
Buisine
Nicolas
N
Chalmers
Ronald
R
D000704:Analysis of Variance; D002118:Calcium / Q000737:chemistry; D002384:Catalysis; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004251:DNA Transposable Elements / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics*; D004305:Dose-Response Relationship, Drug; D008274:Magnesium / Q000737:chemistry; D008345:Manganese / Q000737:chemistry; D008957:Models, Genetic; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D019895:Transposases; D045567:VDJ Recombinases / Q000378:metabolism
D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins; C111652:mariner transposases; D008345:Manganese; D004247:DNA; D019895:Transposases; D045567:VDJ Recombinases; D008274:Magnesium; D002118:Calcium