The Mu transposase tetramer is inactive in unassisted strand transfer: an auto-allosteric effect of Mu A promotes the reaction in the absence of Mu B.
A tetramer of the Mu transposase is the structural and functional core in all three stable higher-order nucleoprotein complexes (Type 0, Type 1 and Type 2 transpososomes) generated in a defined in vitro strand transfer reaction. Although functional in donor cleavage, we report here that contrary to previous belief, the Mu A tetramer is incapable of unassisted strand transfer. The Mu B protein is required to stimulate the tetramer for intermolecular strand transfer. In the absence of Mu B protein we show that additional Mu A molecules must be added to the core tetramer to stimulate intramolecular strand transfer. Mapping experiments indicate that domain II of the assisting Mu A mediates functional interactions with the core tetramer. The recipient site for Mu A stimulated strand transfer on the A tetramer is likely in domain II and is clearly different from the domain IIIb site used by the Mu B protein. The Mu accessory end binding sites and the Mu enhancer are not required in the Mu A assisted strand transfer, suggesting that helper A molecules in solution can interact with the core tetramer to stimulate the reaction. Finally, we argue that the strand transfer activity and protein sites for target interaction reside within the core tetramer; hence the role of the stimulatory A molecules appears to be limited to that of an auto-allosteric effector.
Wu
Z
Z
Department of Biochemistry, The University of Western Ontario, London, Canada.
Chaconas
G
G
D000494:Allosteric Regulation; D010583:Bacteriophage mu / Q000201:enzymology*; D001704:Biopolymers; D004254:DNA Nucleotidyltransferases / Q000737:chemistry / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D019895:Transposases; D014764:Viral Proteins / Q000378:metabolism*
D001704:Biopolymers; D004279:DNA, Viral; D004268:DNA-Binding Proteins; C069215:MuB protein, Enterobacteria phage Mu; D014764:Viral Proteins; D004254:DNA Nucleotidyltransferases; D019895:Transposases