TnpI recombinase: identification of sites within Tn5401 required for TnpI binding and site-specific recombination.
The Bacillus thuringiensis class II transposon Tn5401 encodes a recombinase protein, TnpI, that mediates the resolution of cointegrate molecules generated as intermediates during Tn5401 transposition by the TnpA transposase. This recombination event requires a specific target site, or internal resolution site, at which TnpI binds and catalyzes the exchange of DNA strands. Gel mobility shift assays and DNase I footprinting analyses were used to localize the TnpI binding region to the sequence extending from nucleotides 637 to 747 of Tn5401. Deletions within this region blocked TnpI-mediated recombination in vivo. The 12-bp sequence ATGTCC RCTAAY, present in four copies within the TnpI binding region, is proposed to be the recognition sequence for TnpI binding. TnpI also binds to a single copy of this sequence located within the 53-bp terminal inverted repeats of Tn5401. The unique juxtaposition of recombinase and transposase binding sites at the terminal inverted repeats of Tn5401 suggests that TnpI regulates the binding and/or catalytic activity of TnpA transposase.
Baum
J A
JA
Ecogen Inc., Langhorne, Pennsylvania 19047-3023, USA.
D000595:Amino Acid Sequence; D001413:Bacillus thuringiensis / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D004252:DNA Mutational Analysis; D004254:DNA Nucleotidyltransferases / Q000096:biosynthesis / Q000378:metabolism*; D004251:DNA Transposable Elements / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D009713:Nucleotidyltransferases / Q000378:metabolism; D011485:Protein Binding; D011994:Recombinant Proteins / Q000096:biosynthesis; D011995:Recombination, Genetic; D017384:Sequence Deletion; D019895:Transposases
D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins; D011994:Recombinant Proteins; D004254:DNA Nucleotidyltransferases; D009713:Nucleotidyltransferases; D019895:Transposases; C069598:tnpI recombinase