A1 Refereed original research article in a scientific journal
Mu transpositional recombination: Donor DNA cleavage and strand transfer in trans by the Mu transposase
Authors: Savilahti H, Mizuuchi K
Publisher: CELL PRESS
Publication year: 1996
Journal:Cell
Journal name in sourceCELL
Journal acronym: CELL
Volume: 85
Issue: 2
First page : 271
Last page: 280
Number of pages: 10
ISSN: 0092-8674
DOI: https://doi.org/10.1016/S0092-8674(00)81103-4
Abstract
Central to the Mu transpositional recombination are the two chemical steps; donor DNA cleavage and strand transfer. These reactions occur within the Mu transpososome that contains two Mu DNA end segments bound to a tetramer of MuA, the transposase. To investigate which MuA-monomer catalyzes which chemical reaction, we made transpososomes containing wild-type and active site mutant MuA. By preloading the MuA variants onto Mu end DNA fragments of different length prior to transpososome assembly, we could track the catalysis by MuA bound to each Mu end segment. The donor DNA end that underwent the chemical reaction was identified. Both the donor DNA cleavage and strand transfer were catalyzed in trans by the MuA monomers bound to the partner Mu end. This arrangement explains why the transpososome assembly is a prerequisite for the chemical steps.
Central to the Mu transpositional recombination are the two chemical steps; donor DNA cleavage and strand transfer. These reactions occur within the Mu transpososome that contains two Mu DNA end segments bound to a tetramer of MuA, the transposase. To investigate which MuA-monomer catalyzes which chemical reaction, we made transpososomes containing wild-type and active site mutant MuA. By preloading the MuA variants onto Mu end DNA fragments of different length prior to transpososome assembly, we could track the catalysis by MuA bound to each Mu end segment. The donor DNA end that underwent the chemical reaction was identified. Both the donor DNA cleavage and strand transfer were catalyzed in trans by the MuA monomers bound to the partner Mu end. This arrangement explains why the transpososome assembly is a prerequisite for the chemical steps.