Mechanism of transposition of bacteriophage Mu: structure of a transposition intermediate.
Mu transposition works efficiently in vitro and generates both cointegrate and simple insert products. We have examined the reaction products obtained under modified in vitro reaction conditions that do not permit efficient initiation of DNA replication. The major product is precisely the intermediate structure predicted from one of the current models of DNA transposition. Both cointegrates and simple inserts can be made in vitro using this intermediate as the DNA substrate, demonstrating that it is indeed a true transposition intermediate. The requirements for efficient formation of the intermediate include the Mu A protein, the Mu B protein, an unknown number of E. coli host proteins, ATP, and divalent cation. Only E. coli host proteins are required for conversion of the intermediate to cointegrate or simple insert products. Structures resulting from DNA strand transfer at only one end of the transposon are not observed, suggesting that the strand transfers at each end of the transposon are tightly coupled.
Craigie
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Mizuuchi
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D000255:Adenosine Triphosphate / Q000494:pharmacology; D001426:Bacterial Proteins / Q000502:physiology; D010583:Bacteriophage mu / Q000235:genetics*; D004261:DNA Replication; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004251:DNA Transposable Elements; D004278:DNA, Superhelical; D004279:DNA, Viral / Q000235:genetics* / Q000378:metabolism; D015245:Deoxyribonuclease BamHI; D004706:Endodeoxyribonucleases / Q000378:metabolism; D004720:Endonucleases / Q000378:metabolism; D004926:Escherichia coli / Q000502:physiology; D008274:Magnesium / Q000494:pharmacology; D008957:Models, Genetic; D011142:Polyvinyl Alcohol / Q000494:pharmacology; D011995:Recombination, Genetic; D015719:Single-Strand Specific DNA and RNA Endonucleases; D014764:Viral Proteins / Q000502:physiology
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D004278:DNA, Superhelical; D004279:DNA, Viral; D014764:Viral Proteins; D000255:Adenosine Triphosphate; D011142:Polyvinyl Alcohol; D004706:Endodeoxyribonucleases; D004720:Endonucleases; D004262:DNA Restriction Enzymes; D015245:Deoxyribonuclease BamHI; C035571:endodeoxyribonuclease VII; D015719:Single-Strand Specific DNA and RNA Endonucleases; D008274:Magnesium