title: DNA-promoted assembly of the active tetramer of the Mu transposase.
abstract: A stable tetramer of the Mu transposase (MuA) bound to the ends of the Mu DNA promotes recombination. Assembly of this active protein-DNA complex from monomers of MuA requires an intricate array of MuA protein-binding sites on supercoiled DNA, divalent metal ions, and the Escherichia coli HU protein. Under altered reaction conditions, many of these factors stimulate assembly of the MuA tetramer but are not essential, allowing their role in formation of the complex to be analyzed. End-type MuA-binding sites and divalent metal ions are most critical and probably promote a conformational change in MuA that is necessary for multimerization. Multiple MuA-binding sites on the DNA contribute synergistically to tetramer formation. DNA superhelicity assists cooperativity between the sites on the two Mu DNA ends if they are properly oriented. HU specifically promotes assembly involving the left end of the Mu DNA. In addition to dissecting the assembly pathway, these data demonstrate that the tetrameric conformation is intrinsic to MuA and constitutes the form of the protein active in catalysis.
authors.affiliation: Laboratory of Molecular Biology, NIDDK, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: T A
authors.initials: TA
authors.lastname: Baker
authors.forename: K
authors.initials: K
authors.lastname: Mizuuchi
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D002118:Calcium / Q000494:pharmacology; D004269:DNA, Bacterial / Q000378:metabolism* / Q000528:radiation effects; D004742:Enhancer Elements, Genetic; D007700:Kinetics; D046911:Macromolecular Substances; D008274:Magnesium / Q000494:pharmacology; D008961:Models, Structural; D008969:Molecular Sequence Data; D009713:Nucleotidyltransferases / Q000378:metabolism*; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D011487:Protein Conformation; D011995:Recombination, Genetic; D019895:Transposases; D014466:Ultraviolet Rays
chemical_list: D004269:DNA, Bacterial; D046911:Macromolecular Substances; D009838:Oligodeoxyribonucleotides; D009713:Nucleotidyltransferases; D019895:Transposases; D008274:Magnesium; D002118:Calcium