Transposase makes critical contacts with, and is stimulated by, single-stranded DNA at the P element termini in vitro.
P elements transpose by a cut-and-paste mechanism. Donor DNA cleavage mediated by transposase generates 17 nucleotide (nt) 3' single-strand extensions at the P element termini which, when present on oligonucleotide substrates, stimulate both the strand-transfer and disintegration reactions in vitro. A significant amount of the strand-transfer products are the result of double-ended integration. Chemical DNA modification-interference experiments indicate that during the strand-transfer reaction, P element transposase contacts regions of the substrate DNA that include the transposase binding site and the duplex portion of the 31 bp inverted repeat, as well as regions of the terminal 17 nt single-stranded DNA. Together these data suggest that the P element transposase protein contains two DNA-binding sites and that the active oligomeric form of the transposase protein is at least a dimer.
Beall
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Department of Molecular and Cell Biology, 401 Barker Hall, University of California, Berkeley, CA 94720-3204, USA.
Rio
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D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D002712:Chlorides / Q000494:pharmacology; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004251:DNA Transposable Elements / Q000235:genetics*; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004330:Drosophila / Q000201:enzymology / Q000235:genetics; D006160:Guanosine Triphosphate / Q000494:pharmacology; D017895:Manganese Compounds / Q000494:pharmacology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics; D018411:Spodoptera; D019895:Transposases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D002712:Chlorides; D004251:DNA Transposable Elements; D004277:DNA, Single-Stranded; D017895:Manganese Compounds; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; D006160:Guanosine Triphosphate; D004247:DNA; D019895:Transposases; C025340:manganese chloride