TY - JOUR
T1 - Transpososomes
T2 - Stable protein-DNA complexes involved in the in vitro transposition of bacteriophage Mu DNA
AU - Surette, Michael G.
AU - Buch, Shilpa J.
AU - Chaconas, George
N1 - Funding Information:
We would like to thank Robert Craigie, Kiyoshi Mizuuchi, and Anthony Maxwell for communication of unpublished results and for a copy of their manuscript prior to publication. We are also grateful to Karl Drlica and Josette Rouviere-Yaniv for a copy of their manuscript prior to publication. We would like to thank Dr. E. A. Faust for his generous gift of DNA cellulose, The advice of Diane Moyles and Micky Hall in the prep aration of electron micrographs is gratefully acknowledged and we are thankful to Dr. R. G. E. Murray and the Department of Microbiology and Immunology for use of darkroom and electron microscope facilities. We also wish to thank Barb Green for the typing of the manuscript. This work was supported by grants from the National Cancer Institute of Canada and the Medical Research Council of Canada. G. C. is a recipient of a Scientist Award from the Medical Research Council of Canada and M. G. S. is a recipient of an Ontario Graduate Scholarship.
PY - 1987/4/24
Y1 - 1987/4/24
N2 - We report that two types of stable protein-DNA complexes, or transpososomes, are generated in vitro during the Mu DNA strand transfer reaction. The Type 1 complex is an intermediate in the reaction. Its formation requires a supercoiled mini-Mu donor plasmid, Mu A and HU protein, and Mg2+. In the Type 1 complex the two ends of Mu are held together, creating a figure eight-shaped molecule with two independent topological domains; the Mu sequences remain supercoiled while the vector DNA is relaxed because of nicking. In the presence of Mu B protein, ATP, target DNA, and Mg2+, the Type 1 complex is converted into the protein-associated product of the strand transfer reaction. In this Type 2 complex, the target DNA has been joined to the Mu DNA ends held in the synaptic complex at the center of the figure eight. Supercoils are not required for the latter reaction.
AB - We report that two types of stable protein-DNA complexes, or transpososomes, are generated in vitro during the Mu DNA strand transfer reaction. The Type 1 complex is an intermediate in the reaction. Its formation requires a supercoiled mini-Mu donor plasmid, Mu A and HU protein, and Mg2+. In the Type 1 complex the two ends of Mu are held together, creating a figure eight-shaped molecule with two independent topological domains; the Mu sequences remain supercoiled while the vector DNA is relaxed because of nicking. In the presence of Mu B protein, ATP, target DNA, and Mg2+, the Type 1 complex is converted into the protein-associated product of the strand transfer reaction. In this Type 2 complex, the target DNA has been joined to the Mu DNA ends held in the synaptic complex at the center of the figure eight. Supercoils are not required for the latter reaction.
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U2 - 10.1016/0092-8674(87)90566-6
DO - 10.1016/0092-8674(87)90566-6
M3 - Article
C2 - 3032448
AN - SCOPUS:0023663468
VL - 49
SP - 253
EP - 262
JO - Cell
JF - Cell
SN - 0092-8674
IS - 2
ER -