Identification of base pairs in the outside end of insertion sequence IS50 that are needed for IS50 and Tn5 transposition.
Short DNA sequences at ends of transposable elements are needed as sites for transposition. Previous deletion mapping showed that, in Tn5 and its component IS50 elements, these essential sites are about 19 base pairs long. To determine which positions are important in transposition, we made one or more sequence changes at each position in the IS50 outside (O) end and assayed the effects of these changes on transposition. Our results indicate that the specific base pairs at 18 of the 19 positions are important in transposition. A 9-base-pair segment in the O end corresponds to a binding site for the Escherichia coli DnaA protein. Comparisons of effects of mutations at different positions in this site, and also measurements of Tn5 transposition in dnaA- cells, indicate that DnaA protein participates in O-end-mediated transposition.
Phadnis
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Department of Microbiology, Washington University Medical School, St. Louis, MO 63110.
Berg
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D001426:Bacterial Proteins / Q000502:physiology; D001483:Base Sequence; D001665:Binding Sites; D004252:DNA Mutational Analysis; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics*; D011995:Recombination, Genetic; D013329:Structure-Activity Relationship
D001426:Bacterial Proteins; D004251:DNA Transposable Elements